Author: Augustine Amalraj; Sasikumar Kuttappan; Karthik Varma A C; Avtar Matharu
Date: 2022-11-18 00:00:00+00:00
Keywords: General; Herbs; Technology & Engineering; Spices and Their Roles in Nutraceuticals and Functional Foods; Food Science
Description: Herbs, Spices and Their Roles in Nutraceuticals and Functional Foods gives an overview of the many pharmacological activities associated with herbs and spices, including detailed coverage on their mechanisms and formulations for the food industry. Chapters focus on key ingredients such as Curcuma longa, Piper Nigrum and Trigonella foenum-graecum, with contributors across the globe providing the latest research and advances for each. This is an essential read for scientists who want to understand the fundamental mechanisms behind the bioactive compounds within herbs and spices. The numerous phy
Publisher: Elsevier

Herbs, Spices and Their Roles in Nutraceuticals and Functional Foods

Editors
Augustine Amalraj
Aurea Biolabs (P) Ltd, Kochi, Kerala, India
Sasikumar Kuttappan

R&D Centre, Aurea Biolabs (P) Ltd, Kochi, Kerala, India

Karthik Varma A.C.

R&D Centre, Aurea Biolabs (P) Ltd, Kochi, Kerala, India

Avtar Matharu

University of York, Heslington, United Kingdom

Table of Contents

Cover image

Title page

Copyright

Contributors

Chapter 1. Overview of natural products in nutraceutical industries

1.1. Introduction

1.2. Global nutraceutical market

1.3. Historical perspectives

1.4. Status of natural products–derived nutraceuticals

1.5. Regulation of nutraceutical around the world

1.6. Challenges and future perspectives

1.7. Conclusion

Chapter 2. Curcuma longa

2.1. Introduction

2.2. Phytochemistry and isolation of bioactive components

2.3. Hurdle of bioavailability and matrix design strategies for improving curcumin bioavailability

2.4. Molecular targets of curcuminoids

2.5. Curcuminoids in functional foods

2.6. Different formulations of curcuminoids and their role in nutraceuticals

2.7. Conclusion

Chapter 3. Boswellia serrata

3.1. Introduction

3.2. Use of B. serrata as a nutraceutical or functional food in neurological diseases

3.3. Use of B. serrata as a nutraceutical or functional food in inflammatory diseases

3.4. Use of B. serrata as a nutraceutical or functional food in gastrointestinal diseases

3.5. Use of B. serrata as a nutraceutical or functional food with anticancer properties

3.6. Use of B. serrata as a nutraceutical or functional food in the treatment of diabetes

3.7. Use of B. serrata as a nutraceutical or functional food with antioxidant properties

3.8. Use of B. serrata as a nutraceutical or functional food with antimicrobial properties

3.9. Conclusion and perspectives

Chapter 4. Role of Piper nigrum in functional food and nutraceuticals

4.1. Introduction

4.2. Overview of pepper

4.3. Nutritional and phytonutrient composition of black pepper

4.4. Recommended dosage and quality specifications of P. nigrum

4.5. Functional food and nutraceutical

4.6. Other therapeutic benefits of P. nigrum

4.7. Uses of P. nigrum essential oil

4.8. Culinary and ethnomedical uses of P. nigrum

4.9. Conclusion

Chapter 5. Ocimum sanctum

5.1. Introduction

5.2. Beneficial effects from Ocimum sanctum as a nutraceutical or functional food

5.3. Conclusion and future perspectives

Chapter 6. Role of Moringa oleifera in nutraceuticals and functional foods

6.1. Introduction

6.2. Nutritional value of Moringa oleifera

6.3. Traditional food and potential uses of Moringa oleifera

6.4. Antimicrobial properties

6.5. Anti-inflammatory properties

6.6. Antioxidant properties

6.7. Anticancer properties

6.8. Antidiabetic properties

6.9. Antiasthmatic properties

6.10. Hepatoprotective properties

6.11. Neuroprotective properties

6.12. Conclusion and future directions

Chapter 7. Galangal—Roles in nutraceuticals and functional foods

7.1. Galangal and its phytochemistry

7.2. Galangal in various formulations for different applications

7.3. Biological activities of galangal for different nutraceutical applications

7.4. Conclusion and future prospective

Chapter 8. Bacopa monnieri

8.1. Background

8.2. Plant morphology

8.3. Chemical constituents

8.4. Extraction methods for chemical constituents

8.5. Analytical techniques used for qualitative and quantitative analysis of B. monnieri

8.6. Quantitative analysis of bacosides

8.7. Structure elucidation of saponins in B. monnieri

8.8. Biological activities

8.9. Neuroprotection

8.10. Examples of standardized extract of B. monnieri as herbal formulations

8.11. B. monnieri as a nutraceutical in commercial food products

8.12. Future perspectives

Chapter 9. Trigonella foenum graecum

9.1. Introduction

9.2. Morphology of fenugreek

9.3. Chemical constituents

9.4. Roles of fenugreek as nutraceutical

9.5. Fenugreek as functional food

9.6. Conclusion

Chapter 10. Rosmarinus officinalis

10.1. Introduction

10.2. Rosemary oil and their roles in nutraceuticals and functional foods

10.3. Effect of rosmarinic acid in beverage drinks

10.4. Effect of carnosic acid and rosmarinic acid on cognition

10.5. Effect of ursolic acid

10.6. Conclusion and future perspectives

Chapter 11. Phytochemicals and therapeutic potential of Punica granatum L

11.1. Introduction

11.2. Morphology of Punica granatum

11.3. Nutritional and medicinal importance

11.4. Economic importance

11.5. Phytochemicals present in various parts of pomegranate

11.6. Pharmacological activities of pomegranate

11.7. Role of phytoconstituents in the management of environment pollutant

11.8. Applications of P. granatum in nanotechnology

11.9. Industrial uses of pomegranate

11.10. Patents granted on P. granatum

11.11. Pomegranate in clinical trials

11.12. Toxicity of pomegranate

11.13. Conclusion

List of. abbreviation

Chapter 12. Garcinia cambogia

12.1. Introduction

12.2. Phytochemical analysis

12.3. Hydroxycitric acid

12.4. Pharmacological activities and mechanism of action

12.5. Common adulterants

12.6. Future perspectives

12.7. Declaration of interest

Chapter 13. Camellia sinensis

13.1. Introduction

13.2. Phytochemicals

13.3. Pharmacokinetics

13.4. Pharmacological activities

13.5. Conclusion

Chapter 14. Ferula asafoetida

14.1. Introduction

14.2. Chemical constituents

14.3. Pharmacological and therapeutic potentials of asafoetida

14.4. Toxic effects

14.5. Conclusion

14.6. Conflict of interest

Chapter 15. Concise review on Asparagus racemosus and its role as nutraceuticals and functional foods

15.1. Introduction

15.2. Habit and habitat

15.3. Macroscopic and microscopic characterization

15.4. Cultivation of Asparagus racemosus

15.5. Nutritional and traditional medicinal value of Asparagus racemosus

15.6. Pharmacological activities

15.7. Chemical constituents

15.8. Clinical studies

15.9. Adverse effect and precaution

Chapter 16. Cinnamon as a potential nutraceutical and functional food ingredient

16.1. Introduction

16.2. Species

16.3. Composition of cinnamon

16.4. Natural compounds of cinnamon and therapeutic uses

16.5. Other therapeutic benefits

16.6. Advanced glycation end products

16.7. Culinary uses of cinnamon across the globe

16.8. Conclusion

Chapter 17. Myristica fragrans (Houtt.)

17.1. Introduction

17.2. Taxonomy

17.3. Morphology and pharmacognostical details of M. fragrans fruit

17.4. Estimation of composition in rind, nutmeg, and mace

17.5. Phytochemistry of M. fragrans fruit

17.6. Extraction and analysis of bioactive components from M. fragrans

17.7. Application of M. fragrans

17.8. Focus on future perspective of nutmeg

17.9. Toxicity aspects of M. fragrans

17.10. Conclusion

Chapter 18. “Ashtawarga”: the wonder medicinal herbs of nature

18.1. Introduction

18.2. Home of Ashtawarga

18.3. Lack of standardization of Ashtawarga plants

18.4. Uses of Ashtawarga plants in traditional and modern system of medicine

18.5. Red list category and criteria

18.6. Conservation actions and strategies for future prospects

Chapter 19. Regulatory aspects of nutraceuticals

19.1. Introduction

19.2. Need for regulatory standards

19.3. Overview on different regulatory bodies

19.4. Conclusion

Chapter 20. Current trends and technologies in nutraceutical industry

20.1. Introduction

20.2. Current trends to drive nutraceutical industry: global scenario

20.3. Technologies in nutraceutical industry

Chapter 21. Future prospective of nutraceutical and functional food with herbs and spices

21.1. Introduction

21.2. Traditional nutraceuticals and products

21.3. Health benefits of nutraceuticals and functional foods

21.4. The global nutraceuticals and functional foods market

21.5. Motivators of nutraceutical and functional foods market

21.6. Present status of nutraceutical and functional foods

21.7. Challenges in the nutraceuticals and functional foods market

21.8. Regulatory and perception issues

21.9. A snapshot of global regulations

21.10. Toxicity potential of nutraceuticals and functional foods

21.11. Future prospects of nutraceuticals and functional foods

21.12. Nutraceuticals/functional foods: the future of intelligent food

21.13. Characteristics of success: daring to win

21.14. Conclusion

Index

Copyright

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Contributors

Olufunmilola A. Abiodun,
Department of Home Economics and Food Science, University of Ilorin, Ilorin, Nigeria

Beatrice I.O. Ade-Omowaye,
Department of Food Science, Ladoke Akintola University of Technology, Ogbomoso, Nigeria

Sajad Ahmad Wani,
Islamic University of Science and Technology, Jammu & Kashmir, India

Reem S. Alruhaimi,
Department of Biology, College of Science, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia

Augustine Amalraj,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Prajya Arya,
Islamic University of Science and Technology, Jammu & Kashmir, India

M. Aswini,
School of Sciences, Department of Home Science, The Gandhigram Rural Institute-Deemed to be University, Dindigul, Tamil Nadu, India

Parveen Bansal,
University Centre of Excellence in Research, BFUHS, Faridkot, Punjab, India

Renu Bansal,
Government Medical College, Patiala, Punjab, India

Pamita Bhandari

CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India

Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India

Dinesh Chaudhary,
Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon, South Korea

Victor Antonio Costa Lima,
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

Adegbola O. Dauda,
Department of Home Economics and Food Science, University of Ilorin, Ilorin, Nigeria

Leonardo de Liori Teixeira,
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

Suman Giri,
College of Pharmacy, Daegu Catholic University, Gyeongsangbuk-do, South Korea

Terri Grassby,
Department of Nutritional Sciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford, United Kingdom

Vikas Gupta,
University Centre of Excellence in Research, BFUHS, Faridkot, Punjab, India

Emad H.M. Hassanein,
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University-Assiut Branch, Assiut, Egypt

Md Jawaid Akhtar,
Department of Pharmaceutical Chemistry, College of Pharmacy, National University of Science and Technology, Bousher, Muscat, Oman

J.A.A.S. Jayaweera,
Department of Microbiology, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Colombo, Sri Lanka

George Joseph,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Shintu Jude,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Arya Kadukkattil Ramanunny,
School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, Punjab, India

Gunpreet Kaur,
University Centre of Excellence in Research, BFUHS, Faridkot, Punjab, India

Shah Alam Khan,
Department of Pharmaceutical Chemistry, College of Pharmacy, National University of Science and Technology, Bousher, Muscat, Oman

Bhupinder Kumar,
Department of Pharmaceutical Chemistry, ISF College of Pharmacy, Moga, Punjab, India

Pradyuman Kumar,
Islamic University of Science and Technology, Jammu & Kashmir, India

Neha Kumari,
Islamic University of Science and Technology, Jammu & Kashmir, India

Sachin Kumar Singh,
School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, Punjab, India

Sasikumar Kuttappan,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Gopal Lamichhane,
College of Pharmacy, Wonkwang University, Jeollabuk-do, South Korea

Janeline Lunghar,
School of Sciences, Department of Home Science, The Gandhigram Rural Institute-Deemed to be University, Dindigul, Tamil Nadu, India

Ayman M. Mahmoud

Department of Life Sciences, Faculty of Science and Engineering, Manchester Metropolitan University, Manchester, United Kingdom

Physiology Division, Zoology Department, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt

Mukesh Maithani,
MRU, VCSG Government Institute of Medical Sciences & Research, Srinagar, Uttarakhand, India

Sreejith Mohanan,
New Product Development, Plant Lipids Private Limited, Cochin, Kerala, India

Akhila Nair,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Jithu Nambukulangara Vijayan,
Department of Business Development, Zeon Life Sciences Pvt. Ltd, Sirmour, Himachal Pradesh, India

Samson A. Oyeyinka

Department of Nutritional Sciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford, United Kingdom

National Centre for Food Manufacturing, College of Sciences, University of Lincoln, Lincoln, United Kingdom

Adewumi T. Oyeyinka,
School of Tourism and Hospitality, College of Business and Economics, University of Johannesburg, Bunting Road Campus, Johannesburg, South Africa

Febin Pappachan,
New Product Development, Plant Lipids Private Limited, Cochin, Kerala, India

Vanessa Bonassa Pesse,
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

Ariane Leite Rozza,
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

Nitisha Sendri

CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India

Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India

Amaya Suku,
New Product Development, Plant Lipids Private Limited, Cochin, Kerala, India

A. Thahira Banu,
School of Sciences, Department of Home Science, The Gandhigram Rural Institute-Deemed to be University, Dindigul, Tamil Nadu, India

Ana Laura Tironi de Castilho,
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

C.V. Umesh,
Department of Chemistry, MES Ponnani College, Malappuram, Kerala, India

Bincicil Annie Varghese,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Karthik Varma A.C.,
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Sheetu Wadhwa,
School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, Punjab, India

Chapter 1: Overview of natural products in nutraceutical industries

Abstract

Natural products have been serving as a source of food and nutrition since antiquity. Use of herbs, minerals, and animal-derived and fermented products as nutrients had been passed on from generations to generation as an integral part of diet in human beings. These agents hold a high market share of the modern nutraceutical market as well. Prebiotics, probiotics, herbal formula, dietary fiber, proteins, and other nutrients derived from natural resources are widely available and hold billions of dollar value in the market. Research and development on this field, regulation by regulatory body around the world, and awareness and “back to nature” trend on customer guide this market to another height. So, natural products–derived nutraceutical can shape future nutraceutical market, and nutraceutical industries should focus on research, innovation, and development in this field to fulfill customer demand and hold good nutraceutical market share.

Keywords

Herbal nutraceutical; Natural products; Nutraceutical industries, Regulation of nutraceuticals

1.1. Introduction

“Nutraceutical,” a term coined by Stephen DeFelice in 1989 has been used interchangeably with functional food and dietary supplements these days. He defined nutraceutical as “a food (or part of food) that provides medical or health benefits, including the prevention and/or treatment of disease” [1]. This definition consolidates the concept of nutrition with pharmaceuticals to address the saying of Hippocrates, father of medicine, that “let the food be your medicine.” Nutraceuticals are believed to maintain human health by modifying abnormalities and retaining normal physiological function of the body [2].

The dominance of the market with highly processed food and their implication in health due to nutritional imbalances made humans susceptible to many chronic metabolic diseases. Use of nutraceutical to treat/manage those major health problems such as obesity, cardiovascular diseases, cancer, osteoporosis, arthritis, diabetes, and hyperlipidemia has gained popularity these days. Increased awareness of people toward health coupled with research finding that support the health benefit of these foods in the prevention and/or treatment of diseases has caused the nutraceutical market to grow geometrically. This increased demand, and added value of nutraceuticals has made the food industry and nutritionists to focus on the investigation of disease-preventive and curative potential of food products [3].

Diverse products ranging from herbal formula, isolated nutrients, dietary supplement, genetically modified food, and processed products such as fortified cereals and beverages are produced by the food industry, herbal industry, pharmaceutical, and agribusiness/nutritional conglomerates. Research and development for standardization, product development, and clinical trials are focused by these industries to add health claims to produce an impact on consumer sentiments [4].

Natural products and herbal medicine were used since antiquity for treatment and prevention of diseases [5]. Incorporating several functional foods, spices as part of diet had been practiced since long, which was kept in the shade due to the evolution of the modern diet because of industrialization [6]. Now, those concepts are again bringing attention after clinical evaluation of foods and increased consumer interest in buying food with health claims in it [7,8].

1.2. Global nutraceutical market

Basically, the global market of nutraceutical industries is driven by aging populations, consumer awareness, increasing distribution channels, and increasing healthcare cost. The global nutraceutical market valued around 209 billion in 2017 covering vitamins, dietary supplements, functional drinks, and functional foods (Fig. 1.1). This stake of markets is estimated to reach 373 billion by 2025 [9,10 ]. The United States was believed to hold 75.9 billion worth of nutraceutical market share in 2018 with a compound annual growth rate of 5.3%. Similarly, nutraceutical worth in 2019 and 2020 were $382 and $412 billion, respectively [9,11]. In 2021, the global turnover of the nutraceutical market was valued over $275 billion, which is unpredictable growth of nutraceutical industries that was 7.3$ annual growth. Baby boomers and millennials increased interests in sports and energy, weight loss remedies, natural pharmaceutical alternatives, and the aim of increasing life expectancy were the major players driving this growth [10]. Consumer trend to seek functionality in food and beverages has guided the rapid growth of the nutraceutical market. Growth further increased lately as with the outbreak of corona pandemic, as consumers started looking for solutions that will boost immunity, reduce stress and anxiety, and other health and well-being products [12]. Different marketed nutraceuticals have shown in Table 1.1. Digestive health and beauty-promoting functional food are garnering interest among the USA consumers as supported by increased number of products in market with added prebiotics, probiotics, and collagen [10]. With the increased interest in the medicinal value of marihuana, cannabidiol (CBD) served as the biggest innovation in the nutraceutical industry. The United States market of CBD is expected to grow to 16 billion by 2025 from 2 billion in 2018 [10].

Figure 1.1
Global estimated nutraceutical market share divided into vitamins and dietary supplement, functional drinks and functional food in 2017 (estimated 209 billion totals)[10].

Table 1.1

List of marketed nutraceuticals [9,13–15].

Products | Claimed phytochemicals | Activities
Appetite intercept™ | Caffeine, phenylalanine, and tyrosine | Appetite suppressant
Metabolife ultra caffeine free™ | B-vitamins
Lumatol AC | Cacti
Coral calcium | Calcium and trace minerals | Calcium supplement
Calcirol D-3® | Calcium and vitamins
Ester C | Vitamin | Immune booster and immunomodulator
Chyawanprash | Amla, Ashwagandha, Ppippali
Emergen-C | Vitamin
Celestial healthtone | Dry fruit extract
DanActive® | Lactobacillus casei DN114001 | Probiotics
DanimalsR | Lactobacillus rhamnosus GG (infant diarrhea)
Aciforce, Bacilac, Bactisubtil, Bififlor, Proflora | Freeze-dried probiotics bacteria
Actimel, Activia, Hellus, Jovita probiotisch, Provie, ProViva,Rela, Revital active, Yakult, Yosa, Vifit, Vitamel | Yogurt/drink/dairy products
Ensure fiber | Fiber, digestive health | Prebiotics
Builder's bar | Fiber
Luna bar | Fiber
ZonePerfect shakes | Fiber
Helios nutrition's organic kefir | Bifidogenesis, calcium absorption
Proteinex® | Carbohydrates, minerals, predigested protein, and vitamins | Protein supplements
ThreptinR diskettes | Protein and vitamin
Biovinca™ | Vinpocetine | Neurotonic
Pure red reishi | Ganoderma lucidum | Antidepressant
PNer plus™ | Vitamin and other natural supplement | Neuropathic pain supplement
Olivenol | Natural antioxidant hydroxytyrosol | Dietary supplement
Table Continued

Products | Claimed phytochemicals | Activities
GRD® | Proteins, vitamins, minerals, and carbohydrates | Nutritional supplement
Mushroom optimizer™ | Folic acid, mushrooms, and polysaccharides | Immunity supplement

1.3. Historical perspectives

The distinction between food and medicine begins after times of Hippocrates (460–377 BCE), but the term nutraceutical was coined lately in 1989 by Stephen De Felice. Medicinal practice before consists of wise selection of natural food products. Story about the development and consumption of “elixir vitae”, ‘Ayurvedic literature’s explanation of “Rasayana” are some examples of age-old history representing consumption of nutraceutical for the improvement of health [16,17]. Ayurvedic literature mentions that consumption of Rasayana (nutraceutical) helps to improve metabolic processes like digestion and immunomodulation, mental health, and overall quality of life. It will also biotransform tissues and prevent senility and related diseases. Rasayana are divided into three classes: Kamya (improve liveliness, intellects, etc.), Naimittika (for specific disease conditions), and Ajasrika (for general health). Ajasrika Rasayanas are one which can be consumed on a routine basis for improving quality of life and revitalizing the body [18]. Formulations such as Chyawanprash (for prevention of general health and respiratory complaints), Brahma Rasayana (to protect from mental stress), Phala Ghrita (to maintain reproductive health), Arjuna Ksheerpaka (to protect from cardiovascular complaints), Shatavari Ghrita (to maintain general health of women), and Rasona Ksheerpaka (for cardioprotection) explained in Ayurveda literature are all nutraceuticals derived from natural products and are being used since ages [17].

1.4. Status of natural products–derived nutraceuticals

Primarily, the bioactive compounds are mostly present in fruits, vegetables, grains, etc., at least in small amounts. And depending upon the presence of active constituents, they have diverse therapeutic functions [7,11]. Alkaloids, flavonoids, saponins, tannins, essential oils, anthraquinones, etc., are present herbs including amino acids, vitamins as well as minerals that make them function [5,15,19,20]. Nutraceuticals can be classified into several types based on source, pharmacological activities, chemical properties, and others. Various medicinal plants have been used as herbal nutraceuticals in different formulations (Table 1.2)..

1.4.1. Herbal

Since the prehistoric era, herbal products have been used for prevention, treatment, and mitigation of health, including the maintenance of health, thereby improving the quality of life. Therefore, it has its place in nutraceutical industries because of their culinary and medicinal value. Biologically active phytochemicals such as alkaloids, terpenoids, lignans, saponins, coumarins, polyphenols, flavonoids, etc., from herbs are responsible for regulating physiological activities [7,20]. There are three major categories of herbs relying on their purpose: food herbs (these are herbs consumed as food having normal positive effect on the body and negligible toxicity which can be taken for a long time), medicinal herbs (these categories of herbs are especially used for medicinal purposes in certain medical conditions that have been prescribed by a medical practitioner), and poisonous herbs (these herbs have strong toxic effects either acute or chronic and strictly require skilled clinician for safe handling and effective treatment) [20].

Table 1.2

List of medicinal plants used as herbal nutraceuticals [9,13,20–23].

Scientific name (common name) | Part | Active constituent | Herbal benefits
Allium sativum (Garlic) | Dried bulb | Allin, Allicin | Antiinflammatory, antibacterial, antigout, nervine tonic
Ginkgo biloba (Maidenhair tree) | Leaves | Ginkgolide, Bilobalide | Memory enhancer, antioxidant, platelets aactivating factor antagonist
Zingiber officinale (Ginger) | Rhizomes | Zingiberene, Gingerols | Stimulant, chronic bronchitis, hyperglycemia, throat ache
Echinacea purpurea (Ehinacea) | Whole plant | Alkylamide, Echinacoside | Antiinflammatory, immunomodulator, antiviral
Panax ginseng (Ginseng) | Dried root | Ginsenosides, Panaxosides | Stimulating immune and nervous system
Glycyrrhiza glabra (Liquorice) | Dried root | Glycyrrhizin, Liquirtin | Antiinflammatory, antiallergic, expectorant
Curcuma longa (Turmeric) | Rhizome | Curcumin | Antiinflammatory, antirheumatic, anticancer
Allium cepa (Onion) | Dried bulb | Allicin, Allin | Hypoglycemic, antibacterial, anticholesterol
Aloe barbadensis (Aloes) | Dried leaves | Aloins, Aloesin | Vasodilator, emollient, antiinflammatory, wound healing
Hydrastis canadensis (Goldenseal) | Dried root | Hydrastine, Berberine | Antimicrobial, astringent, antihemorrhagic
Cassia angustifolia (Senna) | Dried leaves | Sennosides | Purgative
Ferula assafoetida (Asafoetida) | Resin | Ferulic acid, Umbellic acid | Stimulant, carminative, expectorant
Aegle marmelos (Bael) | Unripe fruit | Marmelosin | Digestive, antidysentery, appetizer
Centella asiatica (Brahmi) | Whole herbs | Asiaticoside, Madecassoside | Antianxiety, spasmolytic, nervine tonic
Hypericum perforatum (st. john's wort) | Dried aerial part | Hypericin, Hyperforin | Antidepressant, antiviral
Valeriana officinalis (Valeriana) | Dried root | Valerenic acid, Valerate | Analgesic, antispasmolytic
Table Continued

Scientific name (common name) | Part | Active constituent | Herbal benefits
Agave americana (Rambans) | Leaves | Hecogenin, Spirostanol | Antibacterial, laxatives, antiinflammatory
Aloe vera (Ghyukumari) | Leaves | Aloin, Lupeol, Auxin, Alovin | Wound healing, antibacterial, antiulcer
Capsicum annum (Bell pepper) | Fruits | Quercetin, Luteolin | Antioxidant, antiarthritis
Moringa oleifera (Moringa) | Seed | Linoleic acid, Phytate | Antimicrobial, antiinflammatory
Trigonella foenumgraceum (fenugreek) | Whole plant | Trigonelline | Antiinflammatory,
Phyllanthus emblica (Amla) | Fruit | Kaempferol | Antioxidant, antidiabetic
Camellia sinensis (Green tea) | Leaves/bud | Polyphenols | Antiobesity, aanticancer, antioxidant, aantidiabetics
Quercus acutissima (Acorn) | Seed | Tannins, flavonoids, and lignans | Antiinflammatory, antioxidant, aantiobesity, aanticancer
Poncirus Trifoliata (Orange) | Fruits | Flavonoid, Coumarin, terpenoid | Antiallergic, used in gastrointestinal disorder
Rhus verniciflua (lacquer) | Bark | Lutein and sulfuretin | Antiobesity, antioxidant, aanticancer, diaphoretic

1.4.2. Dietary supplements

Dietary supplements are a major part of nutrition, synthesized by incorporating some essential phytochemicals in the diet. Amino acids, fatty acids, minerals, vitamins, and minerals (Table 1.3 and Table 1.4) are considered as functional nutritional factors in dietary supplements [24]. Amino acids are usually found in dietary sources, including eggs, meat, dairy, etc., acting as cofactor and regulating the chemical reaction inside the cell and help to transport nutrients, building blocks of protein, hormones, and neurotransmitters [5,7]. Similarly, minerals help to maintain the bone and cartilage, and improve the nerve impulses. Moreover, inflammatory responses, brain function, and cholesterol levels are regulated by fatty acids like omega-3 [11,24].

Table 1.3

List of resources of minerals found in plants [11,24].

S.No. | Minerals | Fruit sources | Vegetable sources
1 | Copper | Avocado, Litchi, Mango, Pomegranate, Guava, Blackberries | Peas, Potatoes, Pumpkin, Sweet potato, French beans, Spirulina
2 | Iodine | Strawberries, Cranberries, Pineapple, Banana | French beans, Navy beans, Potatoes
3 | Iron | Grapes, Kiwi, Cherries, Dates, Figs, Blackberries | Peas, Potatoes, Spinach, Turnip
4 | Calcium | Orange, Pomegranate, Mulberries, Grapes, Blackcurrant | Amaranth leaves, Broccoli, Beans, Lady's finger, Turnip
5 | Magnesium | Banana, Blackberries, Dates, Guava | Okra, Peas, Beans, Butternut Squash
6 | Potassium | Cherries, Dates, Grapes, Guava, Watermelon, Banana | Bamboo shoots, Pumpkin, Potatoes, French beans
7 | Zinc | Blackberries, Dates, Raspberries, Blackberries | Bamboo shoots, Corn, French beans, Asparagus
8 | Sodium | Apple, Guava, Avocado, Papaya, Mango, Banana | Fresh vegetables
9 | Selenium | Litchi, Mango, Guava, Banana | French beans, Lima beans, Mushrooms, Peas
10 | Phosphorus | Avocado, Litchi, Mulberries, Guava | Maize, potatoes, parsnip, brussels sprouts
11 | Manganese | Banana, Blackcurrant, Grapes | Lima beans, Okra, Peas, Sweet potato

Table 1.4

List of dietary supplements [24].

Dietary supplement | Sources | Significance
Ketogenic diet | Low protein and carbohydrates, high fat | Control seizure
Minimally refined grains | Cereals and grains having calcium supplement | Blood glucose control and antigastrointestinal cancer
Phytoestrogen | Soya flour, linseeds | Regulate estrogen levels and prevent breast cancer
Edible mushrooms | Tonnage, lentinus, Pleurotus | Modulate immune system, antitumor activity, antilipidemic activity

1.4.3. Probiotics

Probiotics are symbiotic microbial organisms (like Bifidobacterium, Lactobacilli, and Eubacteria) that on ingestion ameliorate gastrointestinal (GI) microbial balance [25]. Ideal probiotics should withstand very low pH (1–3) of the stomach, bile salt in the small intestine, and should grow well in the intestinal wall and prevent colonization by pathogenic bacteria [26–28]. They are especially beneficial in inflammatory bowel diseases as they maintain GI health by inhibiting pathogenic infection [25]. They help in the absorption and digestion, stimulating immunity, reducing blood cholesterol, preventing cancer, and playing a role in the synthesis of some vitamins. Due to the huge health benefits of probiotics and awareness of people's healthy food and lifestyle, the probiotics market is worth hundreds of million and is increasing day by day. Fermented dairy products like yogurt and other beverages are natural sources of probiotics [29–31]. Several other formulations of prebiotics are also available in the market. Moreover, encapsulation techniques to prevent probiotics bacteria from a harsh gastric environment have also been developed [32].

1.4.4. Prebiotics

Food ingredients that are beneficial for the growth of probiotics in GI tract such as short-chain carbohydrates (Oligosaccharide) constitute prebiotics. They have the inherent capability of not being digested and/or absorbed in the front part of the GI tract, while can be broken down to beneficial compounds, to be absorbed into the host, by probiotic bacteria in the large intestine. Ideal prebiotics should enhance the growth of beneficial bacteria while should not provide grounds to grow pathogenic ones [33,34]. Studies have shown that prebiotics when consumed together with probiotics reduces the risk of getting stomach and intestinal cancer in adults. Feeding babies with prebiotics like inulin and fructooligosaccharide found to make them less susceptible to cold, diarrhea, and vomiting [19,29,35].

Major prebiotics in use are fructooligosaccharide, lactulose, lactose, stachyose, raffinose, pectin, cellulose, hemicellulose, and xylan. Although many commercially available prebiotics are synthetic, most of them are also found naturally in banana, soybean, garlic, asparagus, artichoke, onion, chicory, etc. Efficiency and amount of those natural prebiotics compounds in these foods can be further improved by employing biotechnological tools. So, manufacturing companies should try to use naturally derived prebiotics such as galactooligosaccharide, inulin, fructooligosaccharide, oligosaccharide from soybean, and others to address consumer preference toward herbal products [29].

1.4.5. Dietary fiber

It is an indispensable part of a healthy diet that helps to efficiently digest and pass out from the digestive system. Mostly, these are composed of carbohydrates and based on their solubility, they are divided into soluble, and insoluble dietary fibers [36,37]. Insoluble dietary fibers are not soluble in water, and therefore, it has been used for GI cancer for rapid passage of food waste from the digestive tract. Similarly, soluble dietary fibers are used to maintain the cholesterol, pressure, and blood glucose level in the body due to having decreased passage of intestinal food waste [20,24,38]. For instance, fruits having pectin and gums are the good sources of dietary fibers.

1.4.6. Proteins

Proteins, structural and functional units of life, are indispensable requirements for the functioning of the body. Previously, most of the world population from developing countries had a deficient protein content in diet [39–41]. Later, the several nutraceutical products are available in different forms such as powder, cookies, etc. Recently, protein-based supplements are available on the basis of each group for children, diabetic patient's groups, hypertensive groups, athlete's groups including the patients with bone problems. For instance, L-arginine amino acid is present in the recaller, and athlete diets because it rapidly metabolizes into nitric oxide and helps to improve muscle development via increasing the blood flow [41,42]. In the diet of thyroid patients, aromatic acid tryptophan has been included to increase thyroid hormone synthesis and maintenance of metabolism [41,43].

1.5. Regulation of nutraceutical around the world

As the nutraceutical market is increasing geometrically, regulation of these products for the protection of consumer health and rights became necessary. Food and Drugs Administration (FDA), an agency under the United States Department of Health and Human Service, allows manufacturers to include “Health Claim” in products, but extensive research to establish that claim (compared to pharmaceutical agent) is not mandatory. These scenarios together with cases of adverse events sometimes posed a considerable risk to the health of consumers. This necessitated strict regulation that ensured quality and safety of products before being marketed [18]. Keeping this scenario in the notice, “Codex Alimentarius” was formulated in 1962 by Food and Agriculture Organizations and World Health Organizations to harmonize the regulatory aspect of food and nutraceutical throughout the world [44]. Table 1.5 highlights different regulations enforced by several countries for the regulation of nutraceuticals products.

Table 1.5

Regulation of nutraceutical and functional food around the world [18,44].

Country/territory | Name of regulation
USA | Dietary Supplement Health and Education Act, 1994
China | State Food and Drug administration of China (SFDA), 2003
Japan | FOSHU (Food for specified Health Uses), 1991
France | French Act on regional Health Governance
Korea | The Health/Functional Food Act (2002)
Europe | Functional Food science in europe (FUFOSE), 1996 Regulation
India | The Food Safety and Standards Act (FSSA), 2006
Canada | Natural Health Products Directorate (NHPD), 2003
Australia | Australian guidelines for complementary Medicines
Nepal | Dietary Supplement guideline, 2016
Taiwan | Health Food Control Act (HFCA),1999
Philippine | Food Fortification Act, 2000
Malaysia | The Food Act, 1983

1.6. Challenges and future perspectives

Lack of regulation, lack of uniformity of regulation, difficulty in getting health claims for the nutraceuticals due to hectic regulatory issues, and less innovation by nutraceutical companies are the biggest challenges of nutraceuticals industries. It is difficult to obtain health claims in the United States of America, Japan, and Europe, though they have formulated laws for the regulation of nutraceuticals. Ambiguity of information on several aspects such as clinical studies, validation of biological markers, defining efficient dose by dose response curve, and information related to adverse effects made it difficult to develop and market nutraceuticals in the global market. Even if some companies made their way to get health claims, any other nutraceutical companies can use those health claims to manufacture similar products as most of the nutraceuticals are not so unique (free ingredient) and are not regulated by patent right. This spoils the competitive advantage of companies who put their effort to obtain that health claim [45].

Some of the major challenges in nutraceutical industry can be listed as follows.

- 1. Lack of regulation/uniformity of regulation throughout the world.

- 2. Difficulty obtaining health claims for nutraceutical products (due to regulation pertaining to clinical evaluation, biologically active marker identification, dose response effect, adverse effect studies in those products).

- 3. Difficult to obtain intellectual property rights due to the lack of exclusive ingredients in products and production of copy products for those free ingredients bringing unethical competition in the market.

- 4. Focus of food companies in conventional food and traditional method of adding functionalities to food by adding vitamin, mineral, and/or herbal extract. Lack of innovation and rigorous research interest in food companies for new product developments.

In spite of many difficulties in nutraceutical industries, it is growing geometrically and holds tremendous potential in the upcoming days.

With the increased interest in the holistic approach of treatment using medical, behavioral, and nutritional intervention, consumer and food companies are now aware about how nutraceutical affects people's health. With increased innovation, technology, and research in the nutraceutical industry, new products will be developed with more efficacy and safety data supported by scientific evidence. Use of biotechnological tools such as genetically modified food is expected to give a new dimension to this industry. Development of personalized food for the target population with defined risk factor, aged population, awareness of healthy lifestyle in developing countries and movement that attract people to live with nature will assure the growth of nutraceutical industries in future [45].

1.7. Conclusion

In conclusion, natural products–derived agents have been serving as an indispensable component of nutraceutical market. New research finding supporting the health benefits of these agents further increased their demands and value. So, natural products–derived nutraceutical can shape future nutraceutical market. Nutraceutical industries should focus on research and development in this field to fulfill customer demand and hold good nutraceutical market share.

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Chapter 2: Curcuma longa

Karthik Varma A.C., Shintu Jude, Bincicil Annie Varghese, Sasikumar Kuttappan, and Augustine Amalraj
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Abstract

Natural phytochemicals derived from plant extracts are used for a wide variety of applications, and research interest is going on a high pace utilizing the efficacy of various bioactive components. Consumer consciousness has led the scientists to deliver value-added products with the incorporation of various plant extracts. Turmeric, the “golden spice” is known from the Vedic cultures where it was used for regular homemade uses and for treating various disorders. It is one of the herbs which finds use from kitchen to clinical studies. Curcuminoids find their application to deliver health benefits, even in different forms. Innovative nutraceutical companies have focused on developing novel technologies to deliver value-added products. The chapter tries to present an overview on the various applications of curcuminoids in functional foods, nutraceuticals, molecular targets, and innovative technologies.

Keywords

Curcuminoids; Functional foods; Matrix design to increase the bioavailability; Molecular targets; Nutraceuticals

2.1. Introduction

Turmeric is regarded as the golden spice with innumerable health benefits. Turmeric, scientifically known as Curcuma longa belongs to the Zingiberaceae family, genus Curcuma. The crop is cultivated in the tropical and subtropical regions around the world [1]. Turmeric originates from the Indian subcontinent and South East Asian countries. Turmeric has been used in various traditional systems of medicine like Ayurveda, Siddha, and Chinese medicine for the treatment of various diseases. The diverse applications of turmeric include use in dietary spice, foods, textile industries, medicinal and nutraceutical fields [2].

The phytochemical analysis of the rhizome had revealed the presence of valuable phytochemical components viz, curcuminoids, volatile oils, proteins, dietary fibers, carbohydrates, and minerals [3]. Curcuminoids form the class of molecules which include curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Depending on the variety, the percentage by weight of curcuminoids contribute to around 3%–8% in turmeric rhizome.

The low toxicity of natural products has opened the way to use in nutraceuticals, dietary supplements, and functional foods and has gained significant popularity globally over the past few decades due to the increased potential health benefits. Most modern nutraceutical industries have focused on developing value-added products which are of natural origin using the incorporation of cutting-edge technologies. The major hurdle in the drug development is designing of the formulation, which has the potential to increase the bioefficacy and often comes as with increased bioavailability of the molecule. There are several promising technologies which are also in commercial use with enhanced pharmacological activities [4]. The numerous health benefits of curcuminoids extend the applications to be used in wide variety of nutraceutical applications [5]. The chapter tries to present an overview on the different phytochemicals, molecular targets, bioavailability, and the role of curcuminoids in various sort of nutraceutical and functional food applications and the various forms with which they are supplemented to the end user (Fig. 2.1).

Figure 2.1
Role of curcuminoids in functional foods and nutraceuticals.

2.2. Phytochemistry and isolation of bioactive components

Investigation of the chemical composition of turmeric has revealed the presence of many valuable phytochemicals such as carbohydrates (70%), moisture (13%), protein (5%), essential oils (phellandrene, sabinene, cineol, borneol, zingiberene, and sesquiterpenes), fat, minerals (potassium, calcium, phosphorus, iron, and sodium), and phenolic curcuminoids (3%–5%) [6]. The curcumin molecule was first isolated by Vogel and Pelletier in the year 1815 as they found a yellow coloring principle in C. longa [7]. Curcumin was first characterized by Milobedeska et al. [8] and was first synthesized by Lampe et al. [9].

There are many methods which are adopted globally to isolate the bioactive components from the rhizomes of C. longa. The dried and crushed raw material is subjected to steam distillation or supercritical fluid extraction (SCFE) to extract the volatile oil components [10,11]. Solvent extraction is often employed to isolate the bioactive curcuminoids which include curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Solvent extraction followed by crystallization and purification has been found to be the most common employed method reported for separating curcumin from turmeric. Several polar and nonpolar organic solvents have been used for the same including hexane, ethyl acetate, acetone, methanol, etc.; ethanol being one of the safest polar solvents is used for the extraction of curcuminoids [12]. Other methods, including Soxhlet extraction, ultrasonic extraction, microwave, zone refining, and dipping methods have been tried, among which the Soxhlet, ultrasonic, and microwave extractions are the most commonly used methods [13–17]. Owing to the use of curcuminoids in dietary supplements, scientists are developing extraction methods employing food grade solvents like triacylglycerols to give good yields [18]. Even though the cost of production is on the higher side, SCFE using carbon dioxide (CO_2) is also often employed to extract curcuminoids. The CO_2 method makes use of the difference in the extraction pressure and application of cosolvents like ethyl alcohol/isopropyl alcohol to extract curcuminoids [19].

2.3. Hurdle of bioavailability and matrix design strategies for improving curcumin bioavailability

Even though curcuminoid is a well-researched moiety, the poor bioavailability and low solubility in water limit the efficacy of the molecules when used for clinical indications. The pharmacological potential of curcuminoids is limited mainly due to their decreased bioavailability and poor pharmacokinetic profile (absorption, distribution, metabolism, and excretion) as well as short half-lifetime in the gastrointestinal (GI) tract [20–22]. The curcumin molecule has less stability in various physiological conditions [23].

The two possible mechanisms by which the bioavailability of curcuminoids can be accessed include the delivery systems and excipient systems. In the formal platform, curcuminoids are isolated from its usual environment and then subjected to encapsulation within a suitable delivery system, which is attributed to the increased bioavailability. In the excipient system, curcuminoids are supplemented in the form of capsules or tablets, followed by combination with an excipient food whose composition and structure are carefully designed to enhance the bioavailability of the curcuminoids. In this scenario, the nature of the excipient has to be selected in such a manner, which doesn't affect the stability of the curcuminoid molecules [24].

There are several methods which are often employed to increase the bioavailability of curcuminoids, which include recreation of the natural matrix of turmeric [25], micellar formulation [26], nanocurcumin formulations [27], etc. The increased bioavailability of the molecule can contribute to improved pharmacological activities.

2.4. Molecular targets of curcuminoids

Curcuminoids can impact a diverse range of molecular targets and signaling pathways, which augment the efficacy of the molecule in a wider variety of pharmacological activities (Fig. 2.2). Curcuminoids are found to be effective in controlling the transcription factors like nuclear factor-κB (NF-κB) [28], activated protein-1 (AP-1) [29], signal transducer and activator of transcription (STAT) proteins [30,31], hypoxia-inducible factor-1 (HIF-1) [32], early growth response-1 (Egr-1) [33], and β-catenin [34], but activates other transcription factors such as aryl hydrocarbon receptor (AhR) [35], activating transcription factor (ATF) 3 [36], C/EBP homologous protein (CHOP) [37], electrophile response element (EpRE) [38], peroxisome proliferator-activated receptor-gamma (PPAR-γ) [39], and NF-E2-related factor (Nrf2) [40]. Curcuminoids are also reported to be molecule of interest to the epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans). Curcuminoids inhibit the EGFR signaling by preventing EGFR tyrosine phosphorylation and suppressing EGFR gene expression that is mediated by the activation of PPAR-γ [41]. Curcuminoids are also reported to have inhibitory activity on tyrosine kinase activity of the HER2/neu receptor [42]. Studies have also found that curcuminoids may directly inhibit angiogenesis and also downregulate expression of various proangiogenic growth factors such as vascular endothelial growth factor, fibroblast growth factor, and epidermal growth factor [43]. Curcuminoids also pose to have inhibitory effects on mammalian target of rapamycin [44], mitogen-activated protein kinase (MAPK) [45]. The antiinflammatory potential of curcuminoids is attributed by its modulatory action on the inflammatory cytokines like tumor necrosis factor alpha (TNF-α) [46] and interleukins [47]. Multiple enzymes are responsible for the progression of inflammatory disorders, which are found to be mediated by curcuminoids like cyclooxygenase-2, inducible nitric oxide synthase, 5-lipoxygenase, and phospholipases A2 [48–51]. Curcuminoids are also found to act on apoptosis-related proteins [52], adhesion molecules, etc. [53].

Figure 2.2
Molecular targets of curcuminoids.

2.5. Curcuminoids in functional foods

The distinct color, flavor, and health benefits pave the way for the use of turmeric and curcuminoids in a wide variety of functional foods and nutraceutical applications. The health awareness among the consumers has led researchers and industries to think on developing a convenient and effective delivery platform for functional foods. Scientific studies have also focused on the use of natural bioactive ingredients in functional food applications.

2.5.1. Cereal-based foods

Cereal-based products such as bread, pasta, cookies, and cakes are often preferred by most people around the world as the main source of energy and nutrients. While fungal growth is a major problem in cereal-based food products, the interest in bakery products continues to rise day by day on account of their nutritional properties [54,55]. Plant-derived bioactive components such as proteins, grape seed extract, fruit pomace powder, ginger, and turmeric are used in bakery products to improve the antioxidant capacity [56–58]. Studies showed that incorporation of raw turmeric powder (8 g) increased antioxidant activity but failed in rheological properties of the cake. Further evaluation by the authors showed that 6g of turmeric powder showed some promising results on all the aspects including the sensory evaluation [59]. Turmeric flowers containing p-cymen-8-ol was incorporated in cookies, and the antioxidant potential was evaluated. Incorporation of higher dosage of the material resulted in a higher degradation rate, which can be attributed to the higher time involved in the cooking of the material [60]. Phytosterol-curcumin–incorporated bread and biscuits showed substantial decrease of cholesterol and risk of cardiovascular disease effects as shown by Ferguson et al. [56]. Incorporation of curcumin in the crackers showed an increase in the antioxidant potential and phenolic contents as well as the bioaccessibility, flavor, and taste of crackers [61]. Curcumin can also be incorporated in many other value-added forms to the cereal foods. Encapsulated curcumin when incorporated in the cakes showed reduction in the hardness as well as chewiness, and a homogeneous yellow color was visibly seen [54]. The antimicrobial activity of curcumin when delivered in the encapsulated form resulted in reducing the growth of the mold spores [62]. While 5mg curcumin per gram were added to wheat flour to make cookies, the results showed that incorporation of curcumin into the bread showed a high antioxidant capacity [55]. Curcumin-enriched turmeric powder extracts were used as a natural antioxidant in the preparation of biscuits along with carrot and grape [63]. These applications showed curcumin when incorporated in baked foods can have a positive impact on the health aspects.

2.5.2. Dairy-based product

Turmeric and its derived phytochemicals are also found to be a promising candidate to be incorporated in dairy-based products. Functional ingredients are commonly used in the development of functional dairy products that include vitamins, probiotics or prebiotics, bioactive compounds, minerals, etc. [64]. The diverse range of pharmacological activities of curcumin has led to the incorporation of turmeric in most of the dairy-based formulations. Milk and milk products containing triglycerides were found to be one of the fine-tuned mediums for the addition of curcumin. The triglycerides present in milk can increase the bioefficacy of curcumin. Curcumin, when incorporated into cow's milk showed good nutritional and antimicrobial activity [65], increased calcium content, and antioxidant activity throughout the storage period [66]. Curcumin was formulated in the nanoemulsion form and included in milk. The study showed that incorporation of curcumin minimized lipid oxidation and also effectively released during intestinal digestion [67]. A study on curcumin addition in yogurts also showed increased total phenolic content and radical scavenging activity [68]. Ricotta cheese supplemented with curcumin was found to increase organoleptic behavior, phenolic and antioxidant activity [69]. Studies have focused on the addition of curcuminoids to the other dairy products like ice cream. Borrin et al. added 10–20mg of pure curcumin per gram of ice cream and found that curcumin could be a better alternative to artificial dye, and the prepared functional ice cream showed similar physiological properties compared to control as well [70]. Curcumin, when incorporated in ghee also contributed to its antioxidant potential (Table 2.1) [71].

2.5.3. Other functional foods

In addition to cereal and dairy-based functional foods, phytochemicals are also added in meat, fruit, and vegetable-based products. Curcumin, when added to the lamb meat at a dosage of 100–300mg/g showed significant reduction in the saturated fatty acid content [70]. A similar effect was observed in meat pate where pork fat was found to be reduced by the addition of curcumin hydrogel, and in turn, the lipid oxidation was also found to be decreased [72]. Curcumin when added to atmospheric packaging of fresh lamb sausages showed an improved antioxidant capacity [73]. In another study, it was found that direct incorporation of 5–15mg curcumin/g flour improved the vitamin E and curcumin content without affecting the sensory characteristics of the chicken nugget [74]. A randomized, single-blind study concluded that compared to control, spice-based beverages containing turmeric increased plasma-peptide tyrosin-tyrosin (p-Ptt) and lowered the “desire to eat” and “prospective consumption (quantity of food wanted to it)” in a healthy human [75].

2.6. Different formulations of curcuminoids and their role in nutraceuticals

Curcuminoids and other value-added phytochemicals have been found to be the primary molecules of interest in the natural products industry, which can be used in the nutraceutical arena. The role of nutraceutical supplements with proven clinical background is gaining much importance nowadays.

2.6.1. Inflammatory disorders

Various nutraceuticals have potential antiinflammatory activities, which help in the prevention and treatment of chronic inflammation-associated diseases. The great advantage of nutraceutical supplements in inflammatory disorders is that it can be used as a complementary alternative to antiinflammatory therapeutic drugs, which leads to a reduction in drug dosage and lower adverse effects.

The roles of curcumin containing supplements are well proved for various sorts of inflammatory disorders. Curcumin being an inhibitor of inflammatory cytokines such as interleukins, TNF-α and cyclooxygenase 2 can have a positive impact on reducing the symptoms associated with inflammatory disorders. Rheumatoid arthritis (RA), being an autoimmune disease and acute chronic inflammatory condition leading to the destruction of joints and severe pain [76]. Several curcumin formulations are evaluated for their efficacy in controlling RA. A novel bioavailable curcumin formulation with the recreation of complete turmeric matrix was evaluated for their efficacy in controlling RA. The RA subjects, who received the curcumin product at both low and high doses reported statistically significant changes in their clinical symptoms such as American College of Rheumatology response, visual analogue scale, C-reactive protein, Disease Activity Score 28, erythrocyte sedimentation rate (ESR), and rheumatoid factor values [77]. Similar study which included curcumin with turmeric oil formulation also showed positive clinical indications on relieving the symptoms associated with RA [78]. Curcumin supplementation at a dosage of 1200mg was supplemented in comparison with phenylbutazone (300mg) for a duration of 2 weeks showed significant improvement in the joint swelling, morning stiffness, and walking time [79].

Osteoarthritis (OA) being a sort of chronic inflammation affecting is severely painful at the onset and increase in strength if not properly treated. Curcumin formulations in different forms are found to be effective in controlling knee OA. A novel formulation containing curcumin with phospholipids and a water dispersible form of curcumin is found to be effective in decreasing pain and nonsteroidal antiinflammatory drugs, without any major adverse effects [80,81]. The polar extract of turmeric rich in polysaccharides was also evaluated for their role in controlling OA. Turmeric extract 500mg capsule was compared against 375mg glucosamine sulfate supplementation for 42 days and resulted in significant improvement in the symptoms associated with OA [82]. In another randomized clinical trial with two parallel groups, Diclofenac 25 and 250mg curcumin supplied twice daily for 3 months also showed improvement in the clinical symptoms associated with OA in the curcumin group [83]. Curcumin extract is also supplied in higher doses like 1500mg/day to evaluate their efficacy in controlling the symptoms associated with OA. Curcumin 1500mg was compared with Ibuprofen 1200mg/day and was supplemented for a duration of weeks [84]. Similarly, many studies with different doses like 0.5–1g was also evaluated for their efficacy in long-term and short-team management of OA [81,85]. The underlying mechanism of curcumin in controlling knee OA was attributed to the downregulation of inflammatory mediators p38MAK, interleukins, vascular cellular adhesion molecule-1, ESR, etc. [86]. Curcumin when used in combination with polyherbs is also found to be effective in controlling many health disorders. Curcumin when used in combination with extracts of Boswellia serrata, Piper nigrum, and Kaempferia galanga is found to be effective in controlling the symptoms associated with knee OA [87].

Supplementation of curcumin is also found to be effective in reducing the inflammatory disorders such as ulcerative colitis, ulcerative proctitis, inflammatory bowel disease, irritable bowel syndrome (IBS), Crohn's disease, etc. The study proved that the supplementation of curcumin at a dosage of 1.1g/day for 1 month, followed by 1.65g/day for 1 month showed significant reductions in the symptoms as well as inflammatory indices in patients [88]. The clinical symptoms associated with ulcerative colitis are also found to be effectively controlled using the supplementation of curcumin/turmeric extract. One study involving 2g/day supplementation of turmeric for a duration of six months showed improvement in the prevention of relapse of disease [89]. Turmeric is also found to be effective in reducing the symptoms associated with IBS, when supplemented with a dosage of 0.072–0.144g/day for a duration of 8 weeks [90]. Not only curcuminoids but also the dietary fibers derived from the rhizomes of turmeric are found to be effective in controlling the symptoms associated with gut health disorders. A novel formulation containing turmeric fiber, curcuminoids, and asafetida extract are found to be effective in relieving the symptoms related to ulcerative colitis as proved in a rat study [91]. The same formulation was proved clinically for their effectiveness in the management of gut health. The study proved that the formulation is effective to increase beneficial bacterial growth such as fecal bifidobacteria and lactobacilli count, the reduction in GI symptoms rating scale, and the study also reported ideal stool form indicated an improvement in gut health [92]. Clinical trials with the turmeric/curcumin supplementation are found to be effective in controlling the symptoms associated with different conditions such as chronic anterior uveitis [93], recurrent anterior uveitis [94], postoperative inflammation [95], gastric ulcer [96], peptic ulcer [97], etc.

2.6.2. Cancer

Cancer being a multistage process involves a series of events and resulting from the dysregulation of genes at multiple steps in cell signaling pathways [98]. The first indication of curcumin's anticancer activities in human participants was shown by Kuttan and coworkers in 1987 and found that topical application of curcumin was found to produce remarkable symptomatic relief as evidenced by reductions in smell, itching, lesion size, and pain [99]. Colorectal cancer being the second leading cause of cancer deaths was found to be effective in controlling many of the biomarkers. A dose escalation study involving 0.036–0.18g/day of curcumin was found to reduce the glutathione (GSH) S-transferase activity in subjects with advanced colorectal cancer [100]. In another study, subjects were given curcumin capsules at three different doses (3.6, 1.8, and 0.45g/day) for 7 days and found that curcumin supplementation reduced the prostaglandin E2 levels [101]. Curcumin/standardized turmeric extract supplementation also reduced the M_1G levels [102], aberrant crypt foci [103], improved body weight, reduced serum inflammatory biomarkers, and induced p53 protein expression [104]. The supplementation of curcuminoid is also found to be effective in pancreatic cancer by reducing the lipid peroxidation [105], mild anticancer activities [106], etc. Supplementation of turmeric extracts standardized to curcuminoids is also found to be effective in controlling the symptoms and biomarkers associated with pancreatic cancer. In a human study, 1.5g/day of curcumin for a duration of six weeks is found to reduce the lipid peroxidation and increase the GSH content in the subjects [107]. The feasibility and tolerability of the combination of docetaxel and curcumin in patients with advanced and metastatic breast cancer were evaluated in an open-label trial. A dose escalation study found that maximum tolerable dose of curcumin was found to be 8g/day, whereas the recommended dose was 6g/day for seven consecutive days every 3 weeks in combination with a standard dose of docetaxel [108]. Curcuminoids are also found to be effective in cancer diseases such as prostate cancer [109], multiple myeloma [110], lung cancer [111], head and neck cancer [112].

2.6.3. Metabolic disorders

Curcumin is found to be effective in controlling the metabolic disorders. Diabetes mellitus (DM) is a condition in which the person has high concentrations of blood sugar, which in turn produces symptoms of polyuria, polydipsia, and polyphagia. Curcumin's ability to decrease blood sugar levels in human patients was first reported and studied in the year 1972. A male patient who had diabetes was supplemented with 5g of turmeric powder over a period for 16 years, after which his fasting blood sugar reduced from 140 to 70mg/dL. Ingestion of curcumin, along with insulin synergistically reduced the blood sugar level to a greater extend [113]. A study was also done to evaluate the potential of a standardized preparation of curcuminoids (NCB-02) against various oxidative stress and inflammatory markers in patients with type 2 DM. Seventy-two subjects were divided into two groups and were randomly given curcumin, twice a day, atorvastatin (10mg, once a day), or placebo for eight weeks. The study showed that curcumin supplementation significantly improved endothelial function and reduced oxidative stress (malondialdehyde) and inflammatory markers (IL-6, TNF-α, and endothelin-1) in the subjects [114]. Microangiopathy is a disease of the small blood vessels (capillaries), in which the capillary walls become so thick and leading to bleeding, leakage of protein, and slow the flow of blood. A bioavailable curcumin formulation containing curcumin with phospholipids (1g/day) was added as a supplement to the standard treatment for weeks, and study showed that the formulation could serve as a potential for controlling diabetic microangiopathy [115]. A novel polyherbal formulation containing curcuminoids, Salacia extract, amla extract, and gymnema extract was preclinically proved for reducing type-2 diabetes in a dose-dependent manner [116].

2.6.4. Neurodegenerative disorders

Curcuminoid derived from turmeric and analogous found in the curry spice turmeric is one of the most prominent dietary polyphenols that has been investigated for its therapeutic properties including neurodegenerative disorders [117]. Alzheimer's disease is a progressive neurodegenerative disorder, usually affecting people older than age 65 years. A randomized, double-blind, placebo-controlled study in 34 patients with Alzheimer's disease proved that curcumin at two different doses (1 or 4g) showed significant improvement in the clinical biomarkers and an increase in the antioxidant activity, as a result of which increased vitamin E content was observed [118]. Dejerine–Sottas disease is often caused by defects in genes of axons and myelin such as myelin P0 (MPZ), peripheral myelin protein 22 (PMP22), PRX, and EGR2. Human study with 1.5g of oral curcumin daily for the first 4 months and 2.5g/day thereafter was supplied to complete a 12-month trial. After 12 months, the patient experienced improvements in the clinical symptoms and the doses were well tolerated [119].

2.6.5. Skin disorders

Turmeric has been used in many traditional systems of medicine, including ancient ayurvedic practices for skin disorders. In South East Asian countries, women apply turmeric to their skin for controlling the skin disorders. The antioxidant property of curcumin was found to be contributing to the therapeutic option for the treatment of vitiligo, which was investigated by the combination of narrow-band ultraviolet B and tetrahydrocurcuminoid cream could result in synergistic therapeutic effects against vitiligo [120]. Another type of skin disorder, psoriasis is a chronic inflammatory skin disease characterized by thick, red, scaly lesions that may appear on any part of the body. The disease affects approximately 2% of the population worldwide and is associated with increased cardiovascular risk [121]. Phosphorylase kinase activity is associated with the depletion of glycogen granules in active psoriasis and an open-label, two-stage clinical trial found that curcumin was well-tolerated, and all participants completed the study with slight improvement in the clinical biomarkers [122].

2.6.6. Other clinical indications

Curcumin being a versatile molecule was studied for a wide range of pharmacological indications. Lupus nephritis recognized as an autoimmune disease, which often can be controlled using curcuminoids, was studied in 24 patients with relapsing or refractory biopsy-proven lupus nephritis and found that curcuminoid has the ability to decrease proteinuria, hematuria, and systolic blood pressure [123]. The role of curcuminoid in renal transplantation was evaluated in 43 subjects with dialysis-dependent cadaveric kidney recipient. Subjects were supplemented with capsules containing curcuminoids (480mg) and quercetin (20mg) to the subjects for a duration of 1 month after surgery. The study concluded that formulation can improve early outcomes in cadaveric renal transplantation, possibly through the induction of heme oxygenase-1 [124]. Abnormalities in oxidative stress and in the antioxidant defense system are commonly reported in patients with β-thalassemia. Even though significant results were not obtained, the study paved the way for the use of curcuminoids in renal diseases [125]. The potential role of curcuminoids is also clinically evaluated in biliary dyskinesia [126], chronic arsenic exposure [127], alcohol intoxication [128], and chronic bacterial prostatitis [129].

2.7. Conclusion

Natural phytochemicals have always gained a lot of accolades for their potential use in nutraceutical and functional food applications. The bioactive components derived from turmeric find extensive application in both nutraceutical and functional food applications. Innovative technologies and innovative delivery platforms can open up the umbrella of application of turmeric-derived components in multivaried applications. Furthermore, the significant improvement in various biomarkers of the clinical studies can open up the research interest of the herb to be used an alternative in comparison to the synthetic counterparts.

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Chapter 3: Boswellia serrata

Ana Laura Tironi de Castilho, Leonardo de Liori Teixeira, Victor Antonio Costa Lima, Vanessa Bonassa Pesse, and Ariane Leite Rozza
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

Abstract

Natural products we usually fed aid in preventing and treating various diseases, besides their nutritional functions. Recent clinical and preclinical studies have shown that consuming herbs and spices triggers beneficial biological effects. Boswellia serrata is a tree found mainly in dry areas in India, and traditional Arab, Ayurvedic, and Chinese medicine use its oleogum resin for medicinal purposes. Furthermore, there are several studies about its role as a nutraceutical and functional food. So, here we reviewed the beneficial effects of B. serrata on nutrition, as it presents antiinflammatory, anticancer, antidiabetic, and antioxidant properties, among others. The highlighting of B. serrata beneficial effects could offer a helpful direction to future studies and the development of a phytomedicine for the prevention and treatment of health conditions.

Keywords

AKBA; Boswellia serrata; Gum resin extract; Oleoresin; Oxidative stress; Pentacyclic triterpenic acids

3.1. Introduction

Boswellia is a small genus with approximately 28 species [1], part of the Burseraceae family, whose members are known for their aromatic essential oils and resins [2]. Boswellia has an oleogum resin internationally known as frankincense or Indian olibanum, widely used in traditional Arab, Ayurvedic, and Chinese medicines to treat some diseases [3]. In addition to the use of frankincense, Boswellia trees can also be fodder or timber [1].

This genus includes Boswellia serrata, a tree with a shrubby appearance, height about 2–6m, and mainly found in dry areas in India [3]. The tree is listed as rare in the Indian Red Data Book, even if it still covers a wide geographic area of India. It is helpful as a commercial B. serrata gum resin extract or frankincense source, and India is the only producer of this oleoresin, mainly in the states Madhya Pradesh, Andhra Pradesh, Gujarat, and Jharkhand [1].

B. serrata gum resin extract comes from incisions in the bark of trunk and branches, emerges a stick-milky liquid, which is coagulated and dried in the oleogum resin. It contains 15%–20% of boswellic, lupeolic, and pentacyclic triterpenic acids [4].

Pentacyclic triterpenic acids can modulate essential pathogenic pathways of inflammatory diseases and cancer, inhibiting crucial pathways to tumor growth and inducing apoptosis in various cancer cell lines [3]. Boswellic acids, such as beta-boswellic acid, keto-beta-boswellic acid, and acetyl-11-keto-beta-boswellic acid (AKBA), are used in the pharmaceutical industry. These compounds have been investigated in the treatment of cancer [1] and inflammatory diseases, as these compounds are able to inhibit the gene expression of proinflammatory cytokines [3].

B. serrata acts as a turpentine source to make paints and varnishes; however, the most important commercial use of B. serrata oleoresin is in Unani and Ayurvedic herbal medicines. It can treat asthma, rheumatoid arthritis, Crohn's disease, osteoarthritis, collagenous colitis, and even fever, pain, and swelling [3]. The European Medicines Agency classified B. serrata gum resin extract as an orphan drug to treat peritumoral brain edema [4] and several cancer types [2]. It also has antiinflammatory properties, inhibiting leukotriene biosynthesis, decreasing proinflammatory cytokine production [5]. It is either used in western biomedicine, religious rituals, perfumery, and cosmetics [1].

B. serrata is also commonly used as a food supplement. Studies about the benefits of B. serrata supplements show their role as nutraceuticals and functional foods [6,7]. In this study, we have investigated the effects of B. serrata as a nutraceutical or functional food, aiming to develop an oral phytomedicine of B. serrata, with several benefits to human health, besides its nutritional value, and decreasing the risk of developing chronic degenerative diseases and being used as an adjuvant in the treatment of several diseases, contributing to the increase of the life quality of people around the world.

3.2. Use of B. serrata as a nutraceutical or functional food in neurological diseases

Stroke is a significant challenge to public health since it has a high incidence and presents a risk of threatening the life [8]. The disease is the third leading reason for death and can lead to disability [9]. A permanent or transient decrease in cerebral blood flow leads to schematic stroke, which represents about 80% [10] and leads to neuronal injury after ischemia-reperfusion (I-R) (Sharma et al., 2014). Oxidative stress plays a vital role in this process, but the body has a natural antioxidant defense mechanism that maintains homeostasis. When this mechanism is disturbed, oxidative stress plays a significant role in I-R and is critical for cerebral damage caused by inflammation and apoptotic cascades [11]. B. serrata resin has antioxidant and antiinflammatory activity, which lead [12] to test the effect of B. serrata aqueous and ethanolic extracts (ABS and EBS, respectively) on cerebral ischemic injury induced by middle cerebral artery occlusion in rats. This occlusion induced the focal cerebral I-R injury, and the animals were randomly distributed into the groups: group 1, the sham one, underwent surgery without passing by ischemia, group 2 received a saline solution and acts as the control group, groups 3–8 received ABS and EBS (125, 250, and 500mg/kg), and group 9 received AKBA (50mg/kg), which is an active triterpenoid compound from B. serrata. All groups received their compounds intraperitoneally injected just after the surgery. The authors evaluated the neurological score of the rats by observing the symmetry of their four limbs; the treatment with ABS or EBS increased the neurological score. The animals’ brains were analyzed, and there was infarction in the ischemic animals compared to the sham group; ABS, EBS, and AKBA treatment remarkably decreased brain infarction. DNA fragmentation in apoptotic cell nuclei was determined using the TUNEL assay. The ischemic animals had increased cell apoptosis, but the animals treated with ABS, EBS, and AKBA showed decreased apoptosis rate compared to the ischemic group. Middle cerebral artery occlusion led to a substantial increase in the expression of cleaved caspase-3 and Bax/BCL-2 ratio, while the treatment with ABS, EBS, or AKBA decreased cleaved caspase-3 expression and Bax/BCL-2 ratio. Lipid peroxidation level was measured through the malondialdehyde (MDA) levels, which was decreased in the animals treated with ABS, EBS, and AKBA. The antioxidant activity was estimated by measuring reduced glutathione (GSH) levels and superoxide dismutase (SOD) activity, and the results showed that ABS, EBS, and AKBA increased GSH levels and SOD activity, evidencing an antioxidant activity. These results showed the neuroprotective potential of B. serrata extracts and its biologically active constituent, AKBA, against focal cerebral ischemia in rats in a dose-dependent manner, probably by decreasing oxidative stress and apoptosis.

B. serrata is used in traditional medicine also because of its properties to improve memory [13], prevent amnesia [14], and other pathologic conditions associated with brain inflammation [15]. However, the molecular mechanism involved in improving memory is still unclear. Long-term memory relies on protein synthesis regulated by transcriptional factors, such as cAMP-response element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) [16]. CREB is involved in many neuronal processes, and it is critical for the conversion of short-term memory to long-term memory [17]. BDNF is a crucial factor involved in memory formation, playing a critical role in various aspects of neural plasticity, which is essential to memory formation [18]. Thereby, Ref. [19] studied the effect of aqueous extract of B. serrata gum resin on the expression of BDNF and CREB genes to clarify the molecular mechanism by which B. serrata affects memory formation. For this, the researchers treated male rats with aqueous extract of B. serrata for 28 days, dividing them into three groups: group 1 was the control, only receiving distilled water, and groups 2 and 3 orally received B. serrata extract 50mg/kg and 100mg/kg. After 28 days of treatment, the researchers tested the animals using the Morris water maze (MWM) to analyze spatial learning and memory, and the results showed that the swimming speed was not different between the groups, which indicates no motor disturbance in the animals. Furthermore, the MWM test showed a difference in escape latency and distance traveled parameters between the groups. About the scape latency, all groups learned to find the hidden platform in the MWM, and it showed a difference between the first and the sixth day of the test. On the sixth day, the scape latency from treated groups had a lower score than the control group, which shows the positive effects of the extract on memory enhancement. The distance traveled of all groups diminished comparing the first with the sixth day, which implies learning appropriately in all groups, and the treated groups showed lower scores than the control on the sixth day, evidencing the positive effect of the treatment. This result implies that B. serrata aqueous extract may positively affect spatial memory. Afterward, the authors performed a quantitative real-time reverse transcription-polymerase chain reaction assay to test the expression of CREB and BDNF genes in the hippocampus, and the results showed no difference in CREB expression between the groups, but increased expression of BDNF transcripts in treated groups. The results showed some enhancements in the rats' spatial memory treated with an aqueous extract of B. serrata that can occur at least partly due to BDNF upregulation. However, since CREB transcription levels were not affected, the BDNF-CREB-BDNF cycle is probably not the pathway affected by the treatment.

Aging can lead to health problems such as cognitive impairment, chronic diseases, physiological disorders, and changes in neurocognitive abilities and memory deficits [20]. Herbal treatment is helpful for diseases such as amnesia and also to reinforce memory. The Iranian traditional medicine recommends Melissa officinalis and B. serrata to improve memory and neurological disorders [21]. Since there were no studies to joint M. officinalis and B. serrata on human memory, Ref. [22] conducted a study to investigate the tablet containing both extracts on older adult memory. The study was a randomized, double-blinded, parallel-controlled trial. A psychologist interviewed the patients to complete Wechsler Memory Scale-Revised (WMS-R) and received both extracts in tablets (two tablets per day during 30 days). The participants completed two questionnaires every week: one with sociodemographic characteristics and the second with WMS-R. There was no sociodemographic difference between the treated and the control group. The total scores of the WMS-R were increased in the treated group, evidencing improved memory in the patients which have ingested the tablets. Specific parameters were also increased in the treated group, such as auditory immediate, visually immediate scores, immediate memory, and working memory. So, M. officinalis and B. serrata oral administration offered beneficial effects on improving memory in older adults, but more research is needed to investigate the efficacy of B. serrata alone in improving memory.

Chemotherapy-induced peripheral neuropathy (CINP) is a major clinical problem associated with cytotoxic agents, resulting in a reduced chemotherapeutic benefit for patients [23]. The evidence of the efficacy from drugs to prevent this side effect is sparse [24]. Some drugs can avoid other types of neuropathies, but cannot avoid CINP. OPERA (GAMFARMA SRL, Milan, Italy) is a dietary supplement capsule combining α-lipoic acid, a potent lipophilic antioxidant [25], B. serrata, which resins contain acids (such as AKBA) with antiinflammatory properties [26], methylsulfonylmethane, which has chemopreventive and antiinflammatory activities [27], and bromelain, the crude extract of pineapple stems, which has antiinflammatory properties [28]. So, Ref. [29] made a prospective study to determine the efficacy and safety of this supplement in patients affected with CINP. The patients were selected with criteria and enrolled at the first clinical sign of neuropathy, during or after chemotherapy, with potentially neurotoxic agents. The patients took OPERA capsules once daily, during 12 weeks. The authors assessed CIPN using evaluation tools, such as the national cancer institute-common toxicity criteria (NCI-CTC), sensory and motor neuropathy score, the clinical version of the total neuropathy score clinical version (TNSc), the modified inflammatory neuropathy cause and treatment, group sensory sum score (mISS), and the visual analog pain scale. All evaluation tools were utilized during the first assessment at the enrollment visit and were repeated every three weeks during 12 weeks by the Chemotherapy Day Unit medical staff. The primary endpoint evaluated the changes of measured scores after 12 weeks of therapy compared to the baseline evaluation done at the beginning of neuropathy. The secondary endpoint evaluated neuropathy reduction at 12 weeks after the beginning of therapy with OPERA. The pain assessment indicated a progressive reduction in pain perceived by patients during the study period. The sensory and motor score showed that the sensory rather than motor component was impaired and that the toxicities incidence diminished at the end of 12 weeks. The mISS scale showed a decreased value of sensory impairment in each evaluation. The TNSc results showed that the median value of the TNSc scale decreased since the enrollment, and the evaluation of individual items showed impairment of the sensory component of CIPN compared with the motor component, and the most affected was vibration. The treatment with OPERA also presented safety, and there was no acute toxicity directly associated with its intake. In conclusion, after 12 weeks of treatment with OPERA, pain and CIPN symptoms showed an improvement. This study has its limited sample size, and nonhomogenous patient population as a challenge, so a prospective controlled trial in a selected population would be interesting to confirm OPERA activity. Moreover, B. serrata was not evaluated alone in improving CIPN symptoms.

3.3. Use of B. serrata as a nutraceutical or functional food in inflammatory diseases

Chronic kidney disease is a progressive disease that can lead to end-stage renal disease or cardiovascular disease [30]. This disease is partly an inflammatory process; acute-phase reactants, oxidative stress, and chronic inflammation are part of its pathological state. Eicosanoids could be involved in the regulation of various of these risk factors, produced via the enzymatic interaction with cyclooxygenases (COXs), lipoxygenases (LOXs), and cytochrome P450 [31]. So is prudent to pursue therapies that intervene in these pathways. Curcumin, a natural product which can decrease prostaglandin E_2 (PGE_2) [32], and B. serrata, which positively affect some human diseases, are two promisor therapies. So, Ref. [33] made a study aiming to report secondary analysis of metabolites downstream of COX and LOX after short-term supplementation with curcumin and B. serrata in early chronic kidney disease in hemodialysis patients. In this regard, the researchers selected about six to eight patients recruited from a community health center in Central Texas to perform a prospective, double-blinded, randomized, placebo-controlled trial. The treatment contained curcumin and B. serrata (two capsules daily) or placebo (roasted rice powder), in addition to their current medications. The patients attended two testing sessions eight weeks apart to collect blood and measure their height, weight, heart rate, blood pressure, and waist and hip circumferences. They performed plasma and serum analysis to the measurement of eicosanoids PGE2, 5-hydroxyeicosatetraenoic acid (5-HETE), 12-hydroxyeicosatetraenoic acid (12-HETE), 15-hydroxyeicosatetraenoic acid (15-HETE), and 13-hydroxyoctadecadienoic acid (13-HODE). As a result of demographics analysis, the placebo and the treatment group differed in height, waist, and hip circumferences without significant differences in age, weight, systolic blood pressure, diastolic blood pressure, resting heart rate, pill compliance, and medication use. They reported only minor side effects during the study. The results showed no difference between groups for baseline levels of 5-HETE, 12-HETE, and 15-HETE, and no differences existed posttreatment for any dependent variables. However, when multivariate analysis was performed, the authors observed an effect for PGE2 between groups and a strong univariate trend toward significance for the interaction between group and time. At the end of the treatment, the treated group had no increased PGE2 levels, while the placebo group had it increased, which means that the supplementation may have attenuated its increase. The interaction of cells involved in the inflammatory process, such as macrophages, lymphocytes, and neutrophils, produces PGE2. The study showed no significant difference in placebo and treated levels of 5-HETE, which conflicts with a report that showed that curcumin diminished its levels in polymorphonuclear leukocytes isolated from blood [34]. The difference in doses charged this difference. There are no reports of other studies examining 12-HETE, 15-HETE, and 13-HODE levels in humans with chronic kidney disease in earlier stages and examining the effects of curcumin and B. serrata. Eight weeks of supplementation have a slight impact on the inflammatory process of chronic kidney disease, but it is necessary more extensive randomized control trials and greater doses of supplements to investigate and elucidate the uselessness of these supplements in mitigating inflammation in the early stage of chronic kidney disease.

Given the formal name LI13109F, the herbal composition containing the extracts of B. serrata gum resin and Aegle marmelos fruit is a promising treatment for asthma. This mix of plant extracts was tested in rats and humans. The extracts decreased airway inflammation in rats, by suppressing the 5-LOX inflammatory pathway and downregulating the expression of the protein Cys-LT1, which is responsible for the infiltration of granulocytes into the bronchoalveolar fluid. The extracts also decreased asthma symptoms in clinical trials, after 14 days of oral treatment. After 56 days of supplementation, asthmatic individuals presented increased levels of serum interferon-gamma and decreased levels of inflammatory marker interleukin-4 without adverse effects. The treatment increased the Th1 response and decreased Th2 response, suppressing the biochemical pathways that cause airway inflammation responsible for the asthma symptoms, leading the authors to conclude that the extract is an effective intervention for airway inflammation [35].

3.4. Use of B. serrata as a nutraceutical or functional food in gastrointestinal diseases

B. serrata is a plant that has been widely studied and used in western medicine due to its pharmacological properties, which are beneficial to the organism. It can treat inflammatory and chronic diseases in extracts or isolated compounds, such as AKBA. However, studies have shown boswellic extracts have decreased bioavailability due to their low permeability and aqueous solubility, impairing their absorption by the animal cells.

Several researchers are trying to find ways to increase B. serrata absorption by the cells. Based on this, Ref. [36] conducted a study aiming to increase the solubility and dispersion of AKBA. The researchers used an ex vivo model that resembles the gastrointestinal tract to increase the intestinal absorption of AKBA, using cyclodextrin and poloxamer solid dispersion formulations. The authors concluded that, when used in proper concentrations, the cyclodextrin complexes act by penetrating the plasma membrane, increasing the permeability of intestinal cells, and allowing better absorption of AKBA. Meanwhile, poloxamer solid dispersion formulation increases the solubility and dispersion of the compound, facilitating its absorption by the cells (increasing it up to 14 times). Both formulations are efficient drug delivery agents and bioavailability enhancers and could improve the absorption of AKBA.

The study conducted by [37] evaluated the efficacy of the administration of B. serrata extract, using a lecithin-based delivery system (Casperome), in the treatment of ulcerative colitis in the low symptomatic remission phase. Among the 43 patients involved, 22 patients daily received B. serrata extract+Casperome supplementation for four weeks; the other 21 patients abstained. The authors found that the supplemented individuals improved their symptoms, such as diffuse intestinal pain, evident and occult blood in the stools, colic, watery stools, malaise, anemia, white blood cell count, the need for concomitant medication, and medical care. Thus, the intake of B. serrata extract+Casperome decreased the need for synthetic pharmaceutical drugs and medical care for patients with ulcerative colitis. Also, no side effects were observed after the ingestion of B. serrata extract+Casperome.

3.5. Use of B. serrata as a nutraceutical or functional food with anticancer properties

B. serrata has also shown anticancer properties, especially in colorectal and breast cancer. A study published by [38] investigated the influence of B. serrata extract on the growth of colon tumors by introducing the extracts into the diet of the rats. Colorectal cancer was induced in these animals by applicating azoxymethane and dextran sodium sulfate, which promoted localized intestinal inflammation, producing intense cell multiplication, thus possibly leading to mutation and the appearance of tumors. Then, the authors offered B. serrata extract supplemented food for 15 weeks to the rats. As a result, there was a decrease in the inflammatory response in the treated group by inhibiting the PI3K/Akt signaling pathway, reducing cell proliferation, and colon tumor growth. So, the researchers concluded that B. serrata extract has a great potential to suppress colon tumorigenesis.

Some researchers have demonstrated the cytotoxic potential of B. serrata against tumor cells using in vitro models [36]. However, further studies with clinical trials in humans and animals are needed to determine its actual effectiveness. As this plant has many different functional and nutraceutical properties, its consumption should be encouraged.

3.6. Use of B. serrata as a nutraceutical or functional food in the treatment of diabetes

The usage of herbs in traditional medicine drives researchers to explore the active ingredients that convey the medicinal properties of these herbs. In this study, considering the stand-alone benefits of the plants Silybum marianum, B. serrata, and Urtica dioica in the treatment of type II diabetes mellitus, a polyherbal treatment comprised the extracts of these three herbs were developed, aiming to merge the beneficial hypoglycemic properties of each single herb therapies, while maintaining safety and minimum adverse effects. The herbal mix decreased serum fasting blood glucose, glycosylated hemoglobin, and triglyceride on the three-month clinical trial alongside the previous hypoglycemic treatment that the patients were already following. Even with these results and previous studies showing the single-herb hypoglycemic benefits, this study showed a potential antihyperglycemic and triglyceride-lowering effect of the herbal formulation, although this beneficial effect could not be attributed only to B. serrata [39].

3.7. Use of B. serrata as a nutraceutical or functional food with antioxidant properties

Intensive exercise triggers advanced glycation end products (AGEs) and oxidative stress markers, which damages healthy tissues. The supplementation of healthy male athletes with curcumin and B. serrata gum resin for three months, together with the Mediterranean diet, decreases AGE and MDA in the athletes' plasma. Soluble receptors for RAGE (sRAGE) and nonesterified fatty acids also decreased after three months of treatment but did not differ from the group of athletes who received only the Mediterranean diet, without the blend of curcumin and B. serrata. The results reported a beneficial effect of supplementation with curcumin plus B. serrata gum resin on glycoxidation and lipid peroxidation in chronically exercising athletes [40].

3.8. Use of B. serrata as a nutraceutical or functional food with antimicrobial properties

Several studies have shown clinical and preclinical B. serrata antimicrobial effects. [41] evaluated the efficacy of a phytotherapeutic combination of L-Methionine associated with Hibiscus sabdariffa and B. serrata to treat acute episodes of urinary tract infections in women. Symptomatic women were treated with two tablets daily or with an antibiotic. After three months, the quality of life was higher in the women treated with the tablets, and one-quarter of the patients from this group presented asymptomatic bacteriuria, while no women treated with antibiotics demonstrated bacteriuria. So, this phytotherapeutic combination, including B. serrata, can improve patients' quality of life, reduce symptoms, and prevent recurrences.

3.9. Conclusion and perspectives

Traditional medicine has used B. serrata since ancient times, as this species has many pharmacological properties when ingested. Preclinical and clinical studies highlighted in this review showed the action of B. serrata in food due to its acids, such as 11-keto-beta-boswellic acid and AKBA. These acids act through different mechanisms and pathways in many kinds of diseases, helping to improve individual health and to combat pathological conditions. Here, we highlighted the importance of developing new researches about this species to elucidate the mechanism behind B. serrata beneficial effect and produce new effective drugs or dietary supplements.

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Chapter 4: Role of Piper nigrum in functional food and nutraceuticals

A. Thahira Banu, and M. Aswini
School of Sciences, Department of Home Science, The Gandhigram Rural Institute-Deemed to be University, Dindigul, Tamil Nadu, India

Abstract

Piper nigrum (black pepper) is used as a spice in modern and traditional cooking; it is also known as King of Spices, among other spices. P. nigrum belongs to the family Piperaceae. In black pepper, the pharmacological property is the presence of specific phenolic components such as alkaloids, flavonoids, carotenoids, terpenoids, etc. It contains flavoring compounds like alkaloids (piperine). Furthermore, pepper is used to treat various diseases such as cough, common cold, sore throat, etc. Many studies reported that piperine present in black pepper possesses several therapeutic properties, that is, antioxidant, antiinflammatory, anticancer, antidiabetic, antimicrobial, antidepressant, enhance the bioavailability, and help to aid digestion. Traditionally, P. nigrum is also used as a culinary ingredient (food), natural preservative (food), flavoring components (cosmetic), and medicine to treat various diseases in pharmacological industries. This chapter mainly focuses on the role of P. nigrum as functional food ingredients and their mechanism of action.

Keywords

Antioxidant; Culinary; Flavonoids; Pharmacological; Piper nigrum; Therapeutic

4.1. Introduction

Piper nigrum (black pepper) is primarily known as the “King of Spices.” Black pepper is the oldest most and widely consumed spice worldwide, and it is native to Southern India. Hence, the primary producer of P. nigrum is Brazil, Malaysia, Indonesia, and India. Pepper, the name derived from the Sanskrit word Pippali and other Indian names are Milagu in Tamil, Kali Mirch in Urdu and Hindi, Peppercorn and Madagascar pepper in English, Kari Menasu in Kannada, Kuru Mulagu in Malayalam, and Miriyam in Telugu [1–3]. Pepper belongs to the family Piperaceae; based on the maturity and ripening stages of P. nigrum fruit, pepper has been classified into three types: white, green, and black pepper. The genus of Piper contains 700 species growing in other parts of the world [4]. It is one of the most prominent genus families and has several traditional medicinal properties.

Black pepper (P.nigrum) is collected from mature fruit, white pepper is obtained from fully ripened (P.nigrum) fruits after removing the outer skin, and the green pepper is prepared from unripe (P.nigrum) fruits. In contrast, green pepper is characterized due to its fresh and herbal flavour. It is responsible for spicy taste due to piperine alkaloids and volatile essential oil [5–7]. Piperine (alkaloid) is found in P. nigrum; it is responsible for the characteristic flavor. Piperine has antioxidant, antimutagenic, anticancer, and antidepressant properties [8]. It is best to enhance the bioavailability of drugs and nutraceuticals. The enhanced bioavailabilities are Co-Q10, β-carotene, curcumin, and antibiotics [9].

P. nigrum was used for various purposes in the human diet as a spice, traditional medicinal agent, preservatives, and biocontrol agent. Whole P. nigrum and its active components are used in various foods and also in medicine. Black pepper is used in different sauces and dishes like meat dishes worldwide [10,11]. In conventional medication, P. nigrum showed gastrointestinal activity such as increased appetite, dyspnea throat diseases, remedies to cough and cold, dysentery, stomachache, discontinuous fever, worms, and piles. It is used as an antipyretic, anti-inflammatory, treats epilepsy and snakebite [1].

Piperine is the primary bioactive compound present in black pepper. It possesses several therapeutic properties, that is, antioxidant, anticancer, antihypertensive, antidepressants, analgesic, antiplatelet, antipyretic agent, and antidiarrheal [1,12–14]. As the most abundant alkaloid present in pepper (piperine content), it is influenced by environmental factors such as origin, climate, and growing conditions. Piperine stimulates the intestinal and pancreatic enzymes, which help to enhance digestion. The therapeutic properties of P. nigrum are the presence of piperine as a part of chemical component and used as a agent.

4.2. Overview of pepper

Taxonomy of black pepper: (Damanhouri and Ahmad, 2014) [3]

Kingdom: Plantae.
Class: Equisetopsida
Superorder: Magnolianae
Order: Piperales
Family: Piperaceae
Genus: Piper
Species: nigrum

P. nigrum quickly grows in the shade with supporting trees or poles. The maximum growth height of climbers is up to 100m, and the diameter of the crown is up to 1.5m. P. nigrum plant had 10–20 essential adventitious roots formed from the base on the mature stem [1]. P. nigrum fruits are small, that is, 3 to 4mm in diameter is known as drupe, whereas unripe, dried fruits black pepper is called peppercorn. The fully ripe fruits (5mm diameter) are dark red, and a single fruit contains one pepper seed. In India, P. nigrum plants start to bear fruits from the fourth to fifth year, and they continue up to seven years. The fresh immature harvested green pepper fruit dried in freeze drier and sundry to form green pepper and black pepper. The red skin of mature ripe fruits was removed, and the seed was sundried to make white pepper [3]. P. nigrum fruit is slightly oval or round. The fruit has been classified into three types: ordinary fruit, ripe fruit (green), and red-orange; the thickness of pepper skin is 1 to 2mm. The immature fruits contain hard skin; ripe fruits have soft skin, whereas red-orange fruits contain juicy skin was easy to peel [15]. In Indian cooking, black pepper has been used since at least 2000 B.C, and it is referred to as “Black Gold.” In Ayurveda, P. nigrum was explicitly mentioned for internal treatment such as fever, gastric, abdominal disorders, and urinary infections, whereas external treatment includes rheumatism, neuralgia, and boils [16]. P. nigrum was used to treat joint pain, muscle pain, and skincare; it improves blood circulation and the respiratory system. However, the bioactive component in black pepper had significant therapeutic impacts on neuromuscular nervous systems and helped to improve digestion. Fig. 4.1 presents the common pepper species used.

4.3. Nutritional and phytonutrient composition of black pepper

P. nigrum (black pepper) is rich in proximate nutrients (carbohydrate, protein, fat), vitamins, and minerals. The nutritional composition of 100 gram of P. nigrum seeds contains (8g) water, (400kcal) energy, (66.5g) carbohydrate, (10g) protein, (10.2g) fat [21], (3.43%–5.09%) total ash, and (10.79%–18.60%) crude fiber [22]. Minerals namely (400mg) calcium, (235.8–249.8mg) magnesium, (1200mg) potassium, and (160mg) phosphorus were found in higher concentration, whereas (10mg) sodium, (17mg) iron, and (1.45–1.72mg) zinc were found in lower concentrations [22,23]. Furthermore, P. nigrum has a crucial vitamin concentration, namely (27.46–32.53mg) vitamin C, (0.74–0.91mg) vitamin B1, (0.48–0.61mg) vitamin B2, and (0.63–0.78mg) vitamin B3 [23].

Figure 4.1
Examples of piper species illustrating the differences between the species. A1 and A2: Piper guineense leaves and seeds [17]; B1: Piper nigrum seeds [17]; B2: Piper nigrum plant [18]; C1 and C2: Piper longum plant and fresh fruit [19]; C3: Piper longum dried fruits [20].

Ashokkumar et al. [24] reported that black pepper contains bioactive components in significant concentration, that is, (2.11–2.80mg) tannin, flavonoids namely (410.0μg) catechin, (56.0μg) myricetin, (13μg) quercetin, and carotenoids such as (260.0μg) lutein and (150.0μg) β-carotene. The essential oil profile of south Indian black pepper seeds is composed of (9.52%–26.95%) β-caryophyllene, (15.13%–20.78%) limonene, (0.00%–19.23%) sabinene, (3.88%–6.48%) α-pinene, (1.32%–7.96%) β-bisabolene, (0.20%–5.57%) α-capaene, (0.18%–4.89%) α-cadinol, and (0.60%–2.94%) α-thujene, respectively [22,25]. Black pepper contains antioxidants, that is, beta carotene, lauric, myristic, and palmitic acids [26].

4.4. Recommended dosage and quality specifications of P. nigrum

The dosage and quality standards of P. nigrum as per FSSAI (Food Safety and Standard Authority of India) standards and AGMARK (Agricultural Marketing) specifications are presented in Table 4.1. P. nigrum is considered Generally Recognized as Safe to use as a flavoring agent in certain foods. It reflects doses are lower than in animal and cell culture studies. Hence, it is not meant to justify the unrestricted use at higher levels. Besides, the approximate intake of P. nigrum in India is about 0.3g/day, a value similar to that of Americans approximate data published by the US Department of Agriculture [28]. The amount is much lower than P. nigrum ingestion in human studies (about 1.5g/d).

Table 4.1

Recommended dosage of Piper nigrum.

Form of the item | Recommended dosage for adults/day and permitted range as per FSSAI, 2016 | AGMARK specifications
Specifications | Bulk density g/1 (Min.) | Moisture % (m/m) (max.) | Total ash % (m/m) (max.) | Nonvolatile ether extract % (m/m). (Min.) | Volatile oil % (ml./100gm) (Min.) | Piperine content % (m/m) (Min.)
Piper nigrum powder | 0.5–2 g | Unbroken pericarp, brown and black color with wrinkled surface, free from added coloring matter and mineral oil | 490–470 | 11.0–12.0 | 6.0–7.0 | 6.0–5.5 | 2.5–2.0 | 4.0–2.0

FSSAI, 2016. FSSAI. Direction for Operationalization of Standards of Coumarin Content in Cinnamon: Notice of Operationalization of Standards of Cinnamon. In Food Security and Safety Authority of India. New Delhi, 2017. https://www.fssai.gov.in/upload/uploadfiles/files/Compendium_Nutra_29_09_2021.pdf.

4.5. Functional food and nutraceutical

The active components in black pepper (piperine) contain numerous biologically active properties, namely antioxidant, antimicrobial, antitumor, antiinflammatory, anticancer, antiobesity, antidepressant activity, and insecticidal activity [1]. Fig. 4.2 presents the biological activity of black pepper/piperine.

4.5.1. Antioxidant property of piper species and P. nigrum

Oxidative stress and lipid peroxidation are known to cause degenerative diseases. The free radicals formed are scavenged easily. Piperine acted as an antioxidant and was proved by many studies [29]. Rauscher et al. [30] studied the antioxidant potential using diabetes mellitus as an oxidative damage model. They investigated using piperine (10mg/kg/day intraperitoneal for four days) on streptozotocin-induced Sprague diabetic rats. Treatment with piperine reversed the diabetic effects on glutathione peroxides and superoxide dismutase (SOD) activities on cardiac, brain, and renal except for liver. Thus, it was concluded that 14 days of subacute treatment with piperine is partially effective. Vijayakumar et al. [31] investigated the antioxidant status of a group of Wistar rats fed with high fat and supplemented piperine. It was evident that a minimal dosage of piperine supplementation along with high-fat to the Wister rats had significant protective action against high-fat diet-induced oxidative stress on the cells.

The spice supplementation reduced the oxidative stress formed due to a high-fat diet. Piperine is used as a bioenhancer in the absorption of curcumin [32,33]. The absorption of curcumin is 2000-fold [34]. Piperine and curcumin were supplemented to patients with COVID-19 as asymptomatic adjuvant therapy and found to reduce mortality and morbidity [35].

Figure 4.2
Biological activity of black pepper/piperine.

Free radical causes damage to the membrane, causing lipid oxidation, less enzyme activity, and causes cancer. Antioxidants entirely stop the process of oxidation. Enzymes involved in the antioxidant protection system are ascorbate, peroxidase, catalase, SOD, which scavenges both radical and nonradical-related oxygen species. The antioxidant activity of piperine can be used to reduce reactive substances, that is, thiobarbituric acid, and maintain glutathione peroxidase, catalase, SOD, glutathione, and glutathione S-transferase. The antioxidant is used to decrease high-fat diet-induced oxidative stress in the cell membrane [3,31]. Oxidative stress is essential for various degenerative diseases, namely immune dysfunction, cancer, diabetes, and Parkinson's. Further, natural antioxidant components form free radicals by scavenging and suppressing degenerative and chronic diseases [1].

4.5.2. Anticancer property of piper species and P. nigrum

Plant secondary metabolites isolated have been widely used as chemopreventive agents [36]. Like most other herbal plants and spices, piper species also contain bioactive compounds to fight malignancy [37,38]. The anticancer property is attributed to the presence of piper amides, and other compounds such as piperine [39], pipernonaline [40], amide alkaloids [41], and piperlongumine isolated in most of the species is most effective compound against cancer cells [42,43]. The same compound also inhibits prostate cancer cells, leukemia, and numerous tumor cell lines [39,44,45]. Piperine and piperlongumine are reported to improve the efficacy of chemotherapeutic drugs [46]. Bezerra [38] conducted a study by administrating pipartine and piperine intraperitoneally for seven days at 50 to 100mgkg/day to female Swiss mice transplanted with sarcoma 180 cells and found that the compounds inhibited tumor proliferation rate. Similarly, Gong et al. [47] studied the efficacy of piperlongumine against human epithelial ovarian cancer cell lines A2780, OVCAR3, and SKoV3. The compound significantly decreases the growth of ovarian cancer cells on 72hr exposure-piperlongumine on OVCAR3 apoptosis at higher concentrations. Sriwiriyajan et al. [48] studied the methanol extract of the P. nigrum fruit against breast cancer MCF-7 and MDA-MB-231 cell lines. Likewise, Prashant et al. [49] reported an ethanolic extract of that fruit showed anticancer activity against three colorectal cancer cell lines (HT-29, HCT-116, and HCT-18) with IC50 values of 7.3, 3.4, and 1.9μg/mL, respectively.

Two bioactive compounds isolated from P. nigrum fruit, namely Kusunokinin and Piperlonguminine, exhibited antiproliferative effects against breast cancer cells, MCF-7, MDA-MB-468, and SW-620. Kusunokinin showed IC50 of 1.18, 1.62, and 2.60μg/mL, respectively. Piperlonguminine showed maximum IC50 activity of 71.54μg/mL MEA-MB-231. The next most common compound of P. nigrum studied for anticancer activity is the piperine and found to be highly effective against breast cancer cells [50]. Similarly, piperine is also effective against prostate cancer cell lines (LNCaP, PC-3, and DU 145) [51]. Selvendiran et al. [52] administrated piperine to the mice-induced lung cancer and found a 95.2% reduction in tumor nodule formalin, serum sialic acid level, and serum gamma-glutamyl transpeptidase activity. Anticancer activity is revealed to the antioxidant activity of piperine and its role in reducing protein extrusion, thus modulating cell proliferation and transformation. Further, black pepper reduces cytotoxicity by modifying enzyme kinetics that controls blood protein lends [53]. The black pepper is reported to have a cytotoxic effect against colon cancer by altering the β-glucuronidase and mucinous activity. The radical scavenging activity of P. nigrum helps arrest the progression of cancer.

Many studies reported that black pepper or piperine helps to inhibit tumor formation. The bioactive component piperine was used to lower the risk of lung cancer by reducing lipid peroxidation and stimulating antioxidative protective enzymes. It has the potential anticancer property [54]. From the mechanism of anticancer activity of piperine against androgen-dependent and independent cells of prostate cancer, it is understood that the proliferation of LNCaP, 2RV1, PC-3, and DU-145 prostate cancer cell dose-dependently was inhibited by piperine. Piperine is nongenotoxic and possesses antimutagenic and antitumor properties. The chemical structure and biological activity of Piper species–related compounds are presented in Table 4.2.

4.5.3. Antiinflammatory property of piper species and P. nigrum

Inflammation is the cause of a few degenerative diseases like Parkinson's, Alzheimer's, and cancer. Inhibition of enzyme activity is also responsible for inflammatory cancer. The antiinflammatory activity was due to the presence of the compound piperine. Mujumdar et al. [65] reported that piperine stimulated the pituitary–adrenal axis and, in turn, indices the antiinflammatory process. Later, conducting in vitro studies said intense activity at dose 20/500μg/mL reduced the level of β-glucuronidase and lactate dehydrogenase. Piperine, along with other bioactive components, inhibits the production of prostaglandins and leukotrienes biosynthesis. The antiinflammatory effect of P. nigrum was determined by Tasleem et al. [66] reported that piperine, along with other bioactive components, inhibits the production of prostaglandins and leukotrienes biosynthesis. Further reported the antiinflammatory activity of piperine was maximum for extracts of piperine from ethanol and hexane and lasted for 120 minutes. The pure piperine showed antiinflammatory activity only for 60 minutes at a dosage of 10 and 15 ml/kg fed to rats induced with inflammation.

In addition, oxidative stress is closely associated to damage caused by excessive production of inflammatory cytokines [67,68], and oxidative stress results in excessive production of proinflammatory cytokines. Further, microglial cells of brain serve as a crucial inflammatory in cell populations in central nervous system. However, activated microglia release a wide range of soluble proinflammatory cytokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) and stimulate neurodegeneration [69,70]. Thus, the proinflammatory cytokines (IL-1β, IL-6, and TNF-α) help to protect against inflammatory damage of ischemic injury [67,68,71–73]. In recent times, numerous studies are indicating the effectiveness of piperine in reducing oxidative stress and, in turn, inflammation and enhancing SOD levels [74,75]. Studies are indicating the use of P. nigrum in treating respiratory inflammatory diseases. Ethanol extracts of P. nigrum were administrated to the ovalbumin-induced asthma mice model, and it decreased the release of IL-4, IL-6, and IL-1β, retinoic acid, TNF-α, and total immunoglobulin E. Further, it decreased fibrous and inflammatory cell infiltration by the cytokines such as Th1, Th2, and Th17. Additionally, the treatment with the extract blocked allergy [76].

4.5.4. Antimicrobial property of piper species and P. nigrum

Extracts of Piper species are effective against both gram-positive and gram-negative bacteria. Further, it is reported that plant microbes do not develop resistance. Piper species contain alkaloids that give increasing antimicrobial activity and do not pose the problem of microbial resistance to antibiotics. Piperine inhibits the growth of Staphylococcus aureus [77]. The alkaloid present in pepper is helpful to act as antiparasitic activity against Giardia and Entamoeba species [78]. P. nigrum or piperine is a natural preservative to protect foods from microbial spoilage. Long-term use of chemical preservatives causes ill effects to the consumer, so they prefer natural, nontoxic products [79]. Rani et al. [80] stated that black pepper or piperine had potential antibacterial and antifungal activity against Escherichia coli, Bacillus subtilis, S. aureus, Aspergillus niger, Alternaria alternate, Aspergillus flavus, and Fusarium oxysporum. Freshly isolated phenolic compounds from black pepper seed help inhibit the growth of S. aureus, S. faecalis, Bacillus, E. coli, and Bacillus cereus [1,81].

Table 4.2

Chemical structures and biological activities of Piper species–related compounds.

Compound/Chemical structure | Species | Biological/pharmacological properties | References

Piperine

P. guineense, P. longum, P. nigrum

Anticancer, antimicrobial, antimalarial

Bezerra et al. [37], Lai et al. [39], Choi et al. [55], Umadevi et al. [56], Niu et al. [57]

Pipernonaline | P. longum, P. retrofractum | Anticancer, antifungal | Lee et al. [40], Muharini et al. [58]

1-[(4E, 9E)-10-(3,4-methylenedioxyphenyl)-4,9-nonadienoyl]pyrrolidine/

P. boehmeriaefolium

Anticancer

Tang et al. [41]

1-[(9E)-10-(3,4-methylenedioxyphenyl)-9-decenoyl]pyrrolidine

P. boehmeriaefolium

Anticancer

Tang et al. [41]

1-[(2E, 4Z,9E)-10-(3,4-methylenedioxyphenyl)-2,4,9-undecatrienoyl] pyrrolidine

P. boehmeriaefolium

Anticancer

Tang et al. [41]

1-[(2E, 4Z,8E)-9-(3,4-mehtylenedioxyphenyl)-2,4,8-nonatrienoyl]pyrrolidine

P. boehmeriaefolium

Anticancer

Tang et al. [41]

Piperarborenine C, D, E | P. arborescens | Anticancer | Tsai et al. [59]
(+)-Arborone | P. arborescens | Anticancer | Tsai et al. [59]
Dihydropiperlongumine | P. guineense, P. nigrum | Anticancer, antimalarial | Scott et al. [60], Rao et al. [61]
Table Continued

Compound/Chemical structure | Species | Biological/pharmacological properties | References

Aduncamide

P. aduncum

Anticancer, antimicrobial

Parmar et al. [4]

Guineensine

P. betle, P. retrofractum

Antimicrobial, anticancer

Muharini et al. [58], Barh et al. [62]

Chabamide

P. chaba, P. retrofractum

Anticancer, antimicrobial, antimalarial

Muharini et al. [58], Rukachaisirikul et al. [63], Ren et al. [64]

4.6. Other therapeutic benefits of P. nigrum

Several studies on spices reported that piperine increases digestion by stimulating pancreatic amylase and decreases the food transit time in the gastrointestinal tract. It also helps in increasing saliva production, gastric secretion, protection, and activation of salivary amylase. P. nigrum and piperine stimulate the liver to secrete bile acid; it plays a vital role in the absorption and digestion of fat. The oral administration of active components, that is, piperine, pipene, piperamines, and piperamides, increases the activity of enzymes such as pancreatic amylase, protease, lipase, and chymotrypsin activity [8,82].

4.6.1. P. nigrum for gastrointestinal health

P. nigrum is used to treat cuts and injuries in traditional medicine. The bioavailability of flavonoid compounds in piperine inhibits the P-glycoprotein and helps in cellular efflux during intestinal absorption [83]. Furthermore, piperine, also known as a natural bioenhancer, interrupts gastrointestinal motility and increases gastric acid secretion [34]. Piperine activates liver, pancreas, and digestive enzymes in the small intestinal mucosa by oral administration. However, the addition of piperine in food flavors and food materials thus increases enzyme activity such as protease, lipase, and pancreatic amylase [8].

4.6.2. Antiobesity property of piper species and P. nigrum

In an antiadipogenesis study of P. nigrum extract and piperine in 3T3-L1 preadipocytes, both the P. nigrum extract and piperine strongly inhibited the adipocyte differentiation of 3T3-L1 cells without affecting cytotoxicity [84]. In another study, black pepper was given to a high fat-fed rat in two doses, that is, 250mg/kg and 500mg/kg and piperines at 20mg/kg were administered for 10 weeks. The plasma and tissue lipid profile significantly reduced total cholesterol, phospholipids, free fatty acids, and triglycerides in both groups. Thus, the results showed that the dietary intake of black pepper or piperine reduces the risk of atherosclerosis via lowering hypolipidemic and antiatherogenic effects [84].

4.6.3. Anxiolytic and property of piper species and P. nigrum

Depression in animals was evaluated via a decrease in sucrose utilization and an increment of immobility time in the tail suspension and forced swim tests. The results showed that in the hippocampus of corticosterone-treated mice, the brain-derived neurotrophic factor protein levels were significantly reduced in the treated mice. Finally, the piperine treatment of mice's behavioral and biochemical changes induced by corticosterone had reverted to normal [85]. The antidepressant activity of the piperine administrated Wister male rats in different doses for 28 days and finally documented that piperine possessed antidepression-like activity and cognitive-enhancing effect during the entire treatment duration [10].

4.7. Uses of P. nigrum essential oil

Essential oil of P. nigrum appears as colorless green liquid, with spicy (pepper) aroma. It has a characteristic odor with a flashpoint of 54°C, specific gravity (0.860–0.884), and refractive index (1.478–1.488) at 20°C [86,87]. The oil extraction was done using various methods, namely liquid carbon dioxide, steam distillation, and solid-phase microextraction. The volatile and nutrient compounds were estimated through column chromatography, gas chromatography, and gas chromatography mass spectrometer [88,89]. Essential oils are recognized as aromatic, odoriferous, and volatile oily liquids extracted from various parts of herbs by steam distillation, expression, etc. It has been proved that some plants’ essential oils have antioxidant, antimicrobial, and also anticancer properties [90–96]. So far, 46 components were identified from pepper and the main compounds such as (E) ocimene, guaiene, (Z, E)-farnesol, guaiol and cadinol. Sabinene, carophyllene, and terpinen-4-ol are present in appreciable quantities. Other imperative sesquiterpene hydrocarbons are bisabolene, cadinenes, calamenene, copaene, cubebenes, ar-curcumene, elemenes, farnesene, guaiene, humulenes, isocaryophyllene, muurolene, santalene, selinenes, ledene, sesquisabinene, and zingiberene [97–99].

The β-caryophyllene is a natural sesquiterpene compound, contribute aroma to the pepper oil, and these compounds have several biological activity properties, that is, antiinflammatory, antibiotic, antioxidant, and local anesthetic [100]. Limonene and Ferreira are other primary components present in black pepper essential oil. These compounds help to protect against colon cancer, gastric cancer, and hepatic cancer and has a promising effect in the prevention of cancer [101,102]. The main antioxidant compound found in essential oil of P. nigrum, namely β-cymene, α-terpineol, α-pinene, β-pinene, δ-3-carene, limoneno, and α-terpineol inhibit bacterial growth [6,103–105]. P. nigrum essential oil, β-caryophyllene (sesquiterpene), limonene, β-pinene, α-pinene, sabinene, 3-carene as well as camphene (mono terpenes) components were identified by suing GC/MS [7]. The major compounds (%) of P. nigrum essential oil are presented in Table 4.3.

Table 4.3

Major compounds (%) of Piper nigrum essential oil.

Compounds | Black | White
E-β-Farnesence | tr.-3.3 | –
Limonene | 16.4–24.2 | 22.6
P-cymene | – | 1.0
Sabinene | 0.1–13.8 | –
α-Copaene | 0.1–3.9 | –
α-Cubebene | 0.2–1.6 | –
α-Humulene | – | 1.0
α-Phellandrene | – | 4.5
α-Pinene | 1.1–16.2 | 4.0
β-Bisabolene | 0.1–5.2 | –
β-Caryophyllene | 9.4–30.9 | 23.4
β-Myrcene | – | 2.7
β-Pinene | 4.9–14.3 | 9.3
δ-3-Carene | tr.-15.5 | 25.2
δ-Elemene | – | 2.1

B.M. Lawrence, Progress in essential oil, Perfum. Flavour. 20 (1995) 49–59; B.M. Lawrence, Progress in essential oils: pepper oil. Perfum. Flavor. 27 (2002) 48–56.

The primary oxygenated terpenes available in the essential oil, that is, caryophyllene oxide, linalool, terpinen-4-ol, and eugenol. The main compounds of P. nigrum essential oil have potential health benefits by (minimizing) pain formed due to tissue injury. Oral administration of essential oil increased SOD, glutathione, and glutathione reeducate enzyme level in the blood of mice, and it provides potential therapeutic applications. Pepper essential oil had antioxidant and antimicrobial properties. The isolated essential oil from pepper can be used as a natural preservative and to preserve orange juice and pork. Moreover, the existing method of food preservation involving the use of artificial/synthetic preservative has undesirable effect on health and also does not completely eliminate microorganism form the preserved foods. So, essential oils are promoted as natural preservatives. Topical application of P. nigrum essential oil (20% in aloe vera gel) was reported to enhance vein visibility and intravenous catheter insertion in the patients [108]. Inhalation of P. nigrum essential oil was able to activate the insular or orbitofrontal cortex, which led to an improved reflexive swallowing movement in older poststroke patients with swallowing dysfunction (dysphagia) in a one-month randomized, controlled study [109]. Moreover, inhaling P. nigrum oil was effective in reducing smoking withdrawal symptoms including cigarette craving and anxiety [110].

4.8. Culinary and ethnomedical uses of P. nigrum

P. nigrum is used mainly as a culinary ingredient in various dishes worldwide; it's preferred for its spicy and hot flavor. Black pepper is mainly used in Western cuisine as a seasoning ingredient and a preservative [111]. In the preparation of soup stocks, soups, curry powders, and paste, whole peppercorns are used. It is also used in stews and soups as a part of a bouquet garni, together with celery, thyme, parsley, and bay leaf. Coarsely crushed peppercorns are added to thick sauces, mayonnaise, or coat fillet steaks or chicken breasts to add hot flavor to food. Ground white peppercorns are mainly used in salads, cream sauces, and light-colored Thai and Chinese cuisine.

Different parts (flower, seed, fruit, and leaf) of P. nigrum are used to manage many ailments. India is one of the countries producing a large quantity of pepper and using it in folk medicines, whereas in European cuisines, it is used only as an ingredient. In traditional folk medicines, P. nigrum is used as paste, decoction, powder, boiled along with milk, or infusions. Almost all ayurvedic formulations will have three ingredients called “Trikatu” or “three acrids” comprising P. nigrum, ginger, and long pepper taken in equal proportions. There are 370 formulations given in the Hand Book of Domestic Medicines and Common Ayurvedic Remedies
; 210 formulations contain “Trikatu” or either one ingredient [112]. The common Ayurveda foods that contain black pepper consumed as medicine are Halu kashaya: Kashaya is a herbal drink recommended for cold and fever and prepared by boiling mint, black pepper, ginger, and jaggery and strained and consumed along with hot milk [113].

Jaljira powder for good digestion and the following ingredients curcumin, black pepper, ginger, mint, and black salt are used to prepare the powder, and this powder can be mixed in water and consumed. Tambuli is prepared and widely consumed in South India, mainly Karnataka. Moreover, this is prepared using any edible green leafy vegetable or pumpkin mixing with cooked vegetables, grated ginger, black pepper, cumin seeds, red chilli, and buttermilk; it is a liquid food [114].

Piperine increases the absorption of selenium, vitamin B12, β-carotene, and curcumin, as well as other compounds. Piperine is shown to possess bioavailability-enhancing activity with various structurally and therapeutically diverse drugs [115]. P. nigrum is used in traditional Ayurvedic medicine due to its active pharmaceutical compound 1-(1-[1, 3-benzodioxol-5-yl]-1-oxo-2, 4 pentenyl) piperidine, which is a pungent nitrogen alkaloid. Furthermore, black pepper is used in cooking some unique flavor foods and as a preservative in food additives or perfumes [116]. Piperine has a low risk of toxicity, is not genotoxic, and does not present any significant adverse effects.

4.9. Conclusion

Black pepper (P. nigrum) contains certain chemical and bioactive compounds, piperine. P. nigrum is also used as a spice and flavoring agent in cooking. Besides, the nutritional composition of P. nigrum includes proximate nutrients, minerals, vitamins, phenolic compounds, flavonoids, terpenoids, and carotenoids. The bioactive compound alkaloid piperine contains primary essential oil such as β-caryophyllene, sabinene, β-bisabolene, limonene, α-pinene, and α-copaene used in food industries as a natural food source, cosmetics industries, and some pharmaceutical industries to enhance therapeutic efficiency. Furthermore, black pepper possesses several medicinal properties, that is, antiinflammatory, antioxidant, antimicrobial, anticancer, aids digestion, and improves the bioavailability of nutrients. Nutraceutical compound (piperine) in black pepper exhibits pharmaceutical properties.

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Chapter 5: Ocimum sanctum

Ana Laura Tironi de Castilho, Victor Antonio Costa Lima, Vanessa Bonassa Pesse, Leonardo de Liori Teixeira, and Ariane Leite Rozza
Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil

Abstract

The ingestion of natural products derived from plants may aid the prevention and treatment of a variety of diseases. Vegetables could act as a nutraceutical or functional food, triggering several beneficial biological effects, proved by clinical and preclinical studies. Ocimum sanctum (Lamiaceae) is an aromatic herbaceous plant characterized by a strong aroma and can be consumed in several ways, such as cold or hot tea, herbal preparations, and as a spice. The therapeutic effects of O. sanctum have been studied, and in this review, we analyzed evidence from the scientific literature about the usage of O. sanctum and its activities as a nutraceutical or functional food. Several effects have been reported, such as adaptogen, neuroprotective, antioxidant, and antihyperglycemic, among others. In conclusion, O. sanctum could be used to improve several human conditions, and more studies should be performed to evidence more and more beneficial effects.

Keywords

Alzheimer's disease; Chronic variable stress; Nutraceutical or functional food; Ocimum species; Lamiaceae; Luteinizing hormone; Tulsi

5.1. Introduction

Ocimum sanctum is a plant that belongs to the Lamiaceae family [1] and is commonly known as holy basil or tulsi. There are approximately 160 species that belong to the Ocimum genus, including O. sanctum, Ocimum americanum, Ocimum basilicum (Ban Tulsi), Ocimum camphora, Ocimum canum (Dulal Tulsi), Ocimum gratissimum (Ram Tulsi), Ocimum kilimandscharicum, Ocimum micranthum, and Ocimum tenuiflorum (Krishna Tulsi). These species have medicinal value and are known as the most sacred plant in India [2].

The Ocimum genus is originally from north-central India, and nowadays, it grows native in the eastern world tropics [3], growing from the Himalayas to the Andaman and Nicobar Islands, in several Arab countries and West Africa. This genus grows in a wide range of soils and climatic conditions and does not demand much care to be propagated by its seeds [2].

O. sanctum is an erect, much-branched annual or biennial subshrub that reaches 30–75cm. It has hairy stems and the leaves are ovate, oblong, obtuse, or acute. The leaves are slightly toothed, opposite, and straightforward, and measure up to 5cm. The flowers are small and purple in elongate racemes in close whorls. The fruits are small and have reddish-yellow seeds [2].

O. sanctum is an aromatic herbaceous plant characterized by a strong and characteristic aroma that occurs mainly because of the volatile oils present in the leaves. The consumption of O. sanctum could be made in various forms; its leaves can be dried and used as an herbal tea in cold or hot preparations, dried powder, or even fresh leaves. It can also be used as a condiment in food preparations, such as salads, and can also be used to produce alcohol tinctures and oil preparations. O. sanctum tastes hot and bitter [3], with an astringent taste. Besides the leaves, its stem, root, and seeds can also produce a formulation that acts systemically and topically [2].

The chemical composition from O. sanctum is very complex. The hydroalcoholic extract of the leaves presents flavonoids as the main constituent, such as ursolic acid, oleanolic acid, orientin, and vicenin [4]. However, many phytochemicals may vary since it has different growing, harvesting, processing, and storage conditions [2]. Most of these chemicals bring beneficial effects for human health, exhibiting diverse pharmacological effects [5].

It explains why this plant is useful in various kinds of traditional medicine, such as Ayurveda, Unani, and Siddha, and is recommended to treat a wide range of diseases, such as malaria fever, bronchitis, catarrh, gastric, cardiac and genitourinary disorders, rheumatism, anxiety, cough, asthma, diarrhea, fever, dysentery, arthritis, eye diseases, otalgia, indigestion, hiccups, vomiting, gastric pain, back pain, ringworm, and skin disorders. The leaves can be used to treat colds, coughs, indigestion, and throat and chest ailments [2,3].

Many clinical and preclinical studies evidence O. sanctum medicinal properties, such as antiinflammatory, antiallergic, immunomodulatory, anticoagulant, antimicrobial, antistress, antiulcer, wound-healing, anticataract, analgesic, antipyretic, antihypertensive, antidiabetic, antifertility, central nervous system depressant, cardioprotective, gastroprotective, hepatoprotective, renoprotective, radioprotective, chemopreventive, and anticancer properties [2,6,7].

The essential oil extracted from the O. sanctum leaves can be applied to alleviate joint pain, and the seeds are helpful for constipation and diarrhea and have anticoagulant properties [3]. The plant can also be an antidote for snakebite and scorpion sting, and the preparation mode for this use is a juice of fresh leaves, flowers, and slender roots [2].

O. sanctum is also used in rituals and practices of daily lifestyle, aiming to promote general health, longevity, and acting prophylactically to prevent diseases. O. sanctum is an adaptogen, because it helps to adapt to stress conditions, helping to deal with the stress of daily life [8], promoting well-being and resilience. O. sanctum also has the property to bring luster to the complexion, sweetness to the voice, fostering beauty, intelligence, and a calm emotional disposition [3].

The phytochemicals and secondary metabolites of plants contribute to their medicinal properties [8], and the presence of antioxidant and antiinflammatory activity is responsible for many O. sanctum physiological beneficial effects. The antioxidant ability protects the body from toxin-induced damage due to its high content of phenolic compounds, which are able to scavenge free radicals [3]. Studies showed that O. sanctum consumption leads to alterations in the natural antioxidant system of the organism, increasing antioxidant molecules levels, such as reduced glutathione (GSH), GSH-related enzymes, such as glutathione peroxidase (GPx), and several antioxidant enzymes, such as catalase and superoxide dismutase (SOD). Antioxidant enzymes protect membranes and cellular organelles from the damage caused by free radical excess [2,3].

Polyphenols present in plants act quenching and neutralizing free radicals because of their properties to donate electrons, which react with free radicals to convert them into more stable products and to terminate radical chain reaction, acting as reducing agents and effectively protecting the biological system from free radical damage under high oxidative stress condition, preventing oxidative stress-related diseases [1,8]. Phenolic compounds from plants help improve food quality, shelf life, and nutritional value. They effectively impede the rancidity of the food due to lipid oxidation, being attractive to researchers that aim to find natural, safe components with better efficiency from plants [8]. In this study, we aimed to investigate, assemble, and discuss the effect of O. sanctum as a nutraceutical or as a functional food in clinical and preclinical studies published in the last seven years.

5.2. Beneficial effects from Ocimum sanctum as a nutraceutical or functional food

O. sanctum has been used in several studies of neurological conditions, such as Alzheimer's disease and depression, discussed as follows. Cognition is an individual psychologic function, which occurs through several functions, such as learning, memory, verbal activity, visual-spatial processing, and psychomotor ability, playing a significant role in daily activities. Dementia is one of the most severe cognitive impairments for aging and middle-aged individuals [9]. Therefore, Ref. [10] investigated the ability of O. sanctum ethanolic extract to enhance cognitive performance, using young and middle-aged rats as the experimental model. The rats were three, six, and nine months old and orally received O. sanctum (50 or 100mg/kg) for 45 days. The results showed that the treatment with O. sanctum (100mg/kg) increased the cognitive ability in the older rats, availed by the radial arm maze test. Also, there was an increase in the density of granular and pyramidal cells in the dentate gyrus, cornu ammonis 1, and cornu ammonis 3 in the hippocampus. There was increased choline acetyltransferase (ChAT) activity, an enzyme that catalyzes acetylcholine synthesis, besides increased gene expression for ChAT. The authors concluded that ingesting O. sanctum enhanced the cognitive ability in middle-aged rats and provided new insight into the role of ChAT in cognitive changes in early to middle-aged life.

Ref. [11] explored the use of O. sanctum in the treatment of chronic variable stress (CVS) paradigm, which is an animal model of depression. Rats were distributed into groups that, apart from the control groups, received different doses of O. sanctum for 16 days after being exposed to CVS. The rats had their weight and immobility time measured during the forced swim test. These measurements showed that O. sanctum was somewhat helpful against the symptoms of CVS, with the immobility time of treated rats being indistinguishable from the control group. Regarding weight variations, rats in the CVS group had a significant body weight loss, while those treated with O. sanctum (200mg/kg) had significant weight gain. The relative weight of adrenal glands and the cortisol levels had an insignificant increase in CVS-induced rats, whereas O. sanctum-treated groups showed a decrease in these aspects. O. sanctum effectively managed stress effects, probably due to inhibiting the release of cortisol by the adrenal glands.

Alzheimer's disease is a progressive neurodegenerative disorder that is the most common type of dementia. This disease mainly affects older adults, impairing cognitive functions, such as memory, language, comprehension, attention, and reasoning. Ref. [12] prospected the antidementia effects of ethanolic extract of O. sanctum and the related mechanisms, using the olfactory bulbectomized (OBX) rodent model of mice, focusing on the cholinergic and vascular endothelial growth factor (VEGF) systems. OBX mice were orally treated with O. sanctum extract (200 or 400mg/kg) or donepezil, starting seven days before the surgery, until day 31 after the surgery, except the three days after the surgery. O. sanctum treatment attenuated OBX-induced impairment of spatial and nonspatial working memories, according to the Y-maze test and novel object recognition test, respectively. O. sanctum treatment also avoided the degeneration of septal cholinergic neurons and the suppression of hippocampal neurogenesis. The oral treatment with O. sanctum decreased the acetylcholinesterase activity and increased VEGF gene and protein expression levels and did not alter the VEGF receptor type 2 gene expression in the hippocampus. These results demonstrated that the administration of O. sanctum could attenuate the cognitive deficits induced by OBX in mice, evidencing an antidementia effect. The hippocampal neurogenesis mediates the effect via protection of central cholinergic systems and restoration of VEGF expression. The authors suggested that the consumption of O. sanctum could act preferentially to prevent the cognitive impairment caused by Alzheimer's disease rather than treating the disease's cognitive dysfunctions.

Ref. [13] also used in vivo and in vitro models of Alzheimer's disease to investigate the effect of ethanolic extract of O. sanctum on the neurons of the dentate gyrus, cornu ammonis 1, and cornu ammonis 3 of the hippocampus. The authors induced Alzheimer's disease in rats through an intraperitoneal injection of trimethyltin, which induces neuronal cell death in the limbic system, especially in the hippocampus. Two groups were orally treated with O. sanctum for seven days before the Alzheimer's disease induction, but one of them also received O. sanctum for 21 days after the induction. Brain samples were collected on days 7, 14, and 21 and analyzed for neuronal density and quantity of neuropeptide Y (NPY), an inductor of neuronal precursor cell proliferation. The neuronal density increased after 14 and 21 days in the group treated with O. sanctum before and after the induction. Nevertheless, the number of neurons decreased in the same period in the group that was only previously treated with O. sanctum. The authors observed the same results for the NPY; a higher quantity in the group that received O. sanctum for 28 days and a decrease in the group that was just pretreated. The authors also used an in vitro model of Alzheimer's disease, treating HEK-293 cells with trimethyltin. O. sanctum treatment reduced the number of apoptotic cells, as indicated by flow cytometry and Hoechst staining. The results showed that ethanolic extract of O. sanctum could act as a neuroprotective agent, as it stimulates the upregulation of NPY, which could prevent apoptosis and stimulate the proliferation of neurons in the dentate gyrus, cornu ammonis 1, and cornu ammonis 3 of the hippocampus.

Ref. [4] aimed to verify the neuromodulatory potential from O. sanctum through experimental research. The authors induced epilepsy in Wistar rats, distributed into groups, and one of them received oral treatment with hydroalcoholic extract of O. sanctum, combined with the drug levetiracetam, aiming to evaluate its pharmacological interactions in the rats. The authors observed that the combination of O. sanctum and levetiracetam greatly improved seizure control and memory retention, besides decreasing oxidative stress and improving the preservation of the neuronal structure. Thus, it was possible to infer that the ingestion of O. sanctum extract works as a viable alternative therapy for the treatment of epilepsy, together with conventional medicine.

Air pollution through cracker smoke produces health problems and some types of cancer, including lung cancer, caused mainly by oxidative stress. O. sanctum is functional due to its innumerable health benefits, which include antioxidant effects. Ref. [14] investigated the protective effect of O. sanctum against cracker smoke-induced lung and brain tissue damage. The researchers found that O. sanctum regulated hematological parameters, such as the quantity of red and white blood cells, blood urea nitrogen, and creatine kinase. O. sanctum supplementation triggered antioxidant activity by decreasing oxidative enzymes such as SOD and catalase, the levels of inducible nitric oxide synthase (i-NOS), and heat shock protein 70 (HSP-70) protein expression. The ingestion of O. sanctum also regulated serotonin, dopamine, and acetylcholine esterase levels, which play vital roles in neuronal function. It preserved the histologic architecture of the lung and brain. These results evidenced the antioxidant and antistress effects of O. sanctum in rats' lungs and brains, protecting from the harmful effects of air pollution.

The antioxidant effect triggered by O. sanctum also offers beneficial effects in the treatment of diabetes. Diabetes mellitus is a metabolic disease characterized by chronic hyperglycemia, leading to decreased natural antioxidant defense mechanisms and increased free radicals due to the auto-oxidation of glucose and proteins. The brain is an essential organ in this disease, since it is involved in mechanisms for sensing and regulating glucose metabolism. Tiwari et al. [15] performed a study to check the biological effect on markers of oxidative stress in brain tissues of diabetic rats treated with the supplementation of composite extracts of leaves (CLEs) of some plants, including O. sanctum. The researchers found higher malondialdehyde (MDA) levels in brain tissue of diabetic rats, evidencing an increased lipid peroxidation. The oral treatment with CLE was able to decrease MDA levels. Diabetic rats showed higher protein carbonyls content in brain tissues, while CLE-treated rats had similar results to the insulin-treated group. The brain tissue of diabetic rats had lower GSH content and lower GPx, SOD, and catalase activity. CLE treatment retained the depletion of GSH and the activity of GPx, SOD, and catalase in the brain. All these results showed that CLE supplementation exhibits potent antioxidant activity, which may protect brain cells from the deleterious effects of hyperglycemia caused by diabetes mellitus. However, this effect cannot be attributed only to O. sanctum, as the rats received a mixture of leaves from different species.

Furthermore, one of the compounds found in this plant, orientin, has been studied by different researchers as a possible treatment for hematological diseases, such as thrombocytopenia, a pathology characterized by a deficiency of blood platelets, very common as a side effect of chemotherapy treatment. So, Ref. [16] performed an assay, introducing O. sanctum leaves in the diet of Wistar rats. The authors exposed the animals to a sublethal dose of radiation to investigate its influence on the production of blood platelets. The ingestion of O. sanctum presented a radioprotective potential, as it accelerated the formation of platelet progenitor cells without interfering in the radiotherapy effect. Furthermore, the supplementation with O. sanctum leaves mitigates the side effects caused by radiotherapy.

Several hormones play a key function in regulating female fertility, and the interplay between the luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone, estradiol, and testosterone controls the reproductive cycle. However, female fertility can be impaired by several factors, leading to infertility. The leaves of O. sanctum have effects on the female hormonal cycle and antifertility effect already reported. The bioactive components found in the plant, such as eugenol, are responsible for those effects. So, Ref. [17] designed a study to test the antifertility potential of O. sanctum and eugenol in female albino rats. The control group received saline (vehicle), the second group received eugenol by daily intramuscular injection, and the third had O. sanctum leaf extract orally administrated. After 15 days, the authors collected the blood and the reproductive tissues. During the experiment, the authors identified the four phases of the estrous cycle (proestrus phase, estrus phase, metestrus phase, and diestrus phase). All the phases had enhanced duration with eugenol administration, increasing the total duration of the cycle. The O. sanctum leaf extract administration showed no effect on the duration of estrous cycle. None of the treatments showed alterations in hormones such as FSH, LH, and prolactin. The testosterone levels decreased in both treatments, indicating destructed ovarian follicles. The progesterone levels increased, leading to a reduced frequency of ovulation. Eugenol-treated animals had enhanced estradiol levels, which represents an alteration in the hypothalamus–pituitary–gonadal axis; O. sanctum had no effect. It was possible to conclude that eugenol and O. sanctum leaf extract administration has antifertility activity, and the effect of O. sanctum is due to the presence of eugenol in O. sanctum leaves.

As the liver and kidneys are widely sensitive to lead exposure, Ref. [18] made a study to evaluate the ability of the ethanolic extract from the leaves of O. sanctum to protect these organs against leaf acetate-induced nephrotoxicity and hepatotoxicity in mice. The researchers used healthy male mice divided into groups that were orally treated by 24 days with tween (vehicle), vehicle and leaf acetate, or O. sanctum leaf extract (140mg/kg, 280mg/kg, or 560mg/kg) plus leaf acetate. After 24 days, the authors euthanized the animals. They used the left lobe of the liver and the left kidney to measure MDA levels, a biomarker of oxidative stress, presenting high levels in the clinical situation. The researchers also measured blood urea nitrogen, creatinine, serum glutamic-oxaloacetic transaminase, and serum glutamic pyruvic transaminase. As a result, all these parameters decreased in the groups treated with O. sanctum. The authors made histopathological analyzes of the right lobe of the liver and the right kidney, using hematoxylin and eosin staining. The control group showed hepatic structural changes, such as hepatic necrosis, in a higher number than the three O. sanctum-treated groups. Healthy kidneys showed normal tubular epithelial cells and glomerulus and significant hydropic degeneration and necrosis prevention, while the control group had massive hydropic degeneration and necrosis. The O. sanctum extract mitigated cell destruction in the kidney. Leaf acetate enhances the intracellular formation of reactive oxygen species, causing liver and kidney damage. However, the treatment with different doses of ethanolic extract from the leaves of O. sanctum prevented this damage due to its antioxidant mechanism, attenuating oxidative stress in the liver and kidney, and showing the practical effect against lead acetate-induced hepatotoxicity and nephrotoxicity in mice.

5.3. Conclusion and future perspectives

Scientific research about O. sanctum shows many psychological and physiological beneficial effects, mainly because of its high content of antioxidant compounds. These compounds act on the natural antioxidant system of the cells, improving their ability to prevent the damage caused by reactive oxygen species. This effect explains the interest in consuming O. sanctum for maintaining health. O. sanctum can be used in various forms, demonstrating different benefits depending on the form of consumption, ingested, as cold or hot teas, spice, and other forms. Since O. sanctum demonstrates many beneficial effects, it is abundant, presents low cost, and its consumption is safe, the need for research to better elucidate its mechanism of action becomes evident for safe use in health and diseases.

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Chapter 6: Role of Moringa oleifera in nutraceuticals and functional foods

Samson A. Oyeyinka
[1]
,
[2]
, Olufunmilola A. Abiodun
[3]
, Adewumi T. Oyeyinka
[4]
, Adegbola O. Dauda
[3]
, Terri Grassby
[1]
, and Beatrice I.O. Ade-Omowaye
[5]

1
Department of Nutritional Sciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford, United Kingdom
2
National Centre for Food Manufacturing, College of Sciences, University of Lincoln, Lincoln, United Kingdom
3
Department of Home Economics and Food Science, University of Ilorin, Ilorin, Nigeria
4
School of Tourism and Hospitality, College of Business and Economics, University of Johannesburg, Bunting Road Campus, Johannesburg, South Africa
5
Department of Food Science, Ladoke Akintola University of Technology, Ogbomoso, Nigeria

Abstract

The Moringa tree is a medicinal plant that has been widely used in traditional medicine and food applications. All the parts of the tree are rich in nutrients that could be harnessed to address food and nutritional challenges in developing nations of the world. This chapter summarizes the role of Moringa oleifera in nutraceuticals and functional foods with emphasis on nutritional and traditional uses, antiasthmatic, antibacterial, anticancer, antidiabetic, antifungal, antiinflammatory, antioxidant, antiviral, hepatoprotective, and neuroprotective properties of the seed, leaf, and flower. While the leaf has been widely used in food applications, the seed and flower have only been moderately used and characterized, thus necessitating further research into these parts of the plant. Future studies are required to fully explore the nutraceutical, functional, and food applications of the M. oleifera plant to enhance its industrial potential.

Keywords

Flower; Functional foods; Leaf; Moringa oleifera; Nutraceuticals; Seed

6.1. Introduction

Historically, the Moringa plant originated from Asian countries, including Afghanistan, Bangladesh, India, and Pakistan [1], but is now widely grown in other regions of the world because it can adapt to a wide spectrum of environments [2]. Taxonomically, Moringa oleifera belongs to the family of Moringaceae [3] of the sole genus Moringa which has 13 species including M. arborea, M. borziana, M. concanensis, M. drouhardii, M. hildebrandtii, M. longituba, M. oleifera, M. ovalifolia, M. peregrine, M. pygmaea, M. rivae, M. ruspoliana, and M. stenopetala [2]. Of these species, M. oleifera Lam. (Moringa) is the most widely cultivated and it is highly nutritious with several uses [4,5].

The Moringa tree is generally called the tree of life or a miracle plant because of its use in medicine and nutrition [5]. Different parts of the tree, such as the seed, leaf, and flower, have been found useful in folk medicine for treating different ailments [6] because they vare rich sources of nutrients, such as amino acids, minerals, protein, and vitamins [7]. The different parts of the Moringa tree are therefore used as dietary supplements in regions of the world where malnutrition is a challenge, especially in children and infants [7]. According to the United Nations, the leaf powder could be used to enrich food with vital nutrients, as such, it could support food and nutrition security, i.e., as a buffer against malnutrition [8]. The plant has been used for a long time in food [5,9], livestock feed, and aquaculture [10–12], and it is resistant to many climatic conditions [13]. Of all the parts of the tree, the leaves are the most nutrient-dense, especially in essential amino acids, vitamins, and various minerals [14,15], which has informed their use as alternative sources of nutrient supplements and growth promoters in most developing countries [16]. Furthermore, it also contains phytochemicals such as phenolic acids and flavonoids, with physiological and pharmacological activities indicative of their functional and nutraceutical roles. Functional foods, according to Kalra [17], are foods that provide the body with health-promoting components in addition to the required amount of nutrients needed for healthy survival. These categories of foods must, however, possess physiological benefits [18]. It is important to note that there are different perspectives to what constitutes a functional food, but essentially, according to the National Academy of Sciences, it must provide health benefits beyond the traditional nutrients it contains [19]. Nicoletti [20] further opined that beyond providing energy and nutrients, functional foods must beneficially regulate one or more targeted functions in the body, by enhancing a certain physiological response and/or by reducing the risk of disease. Given this background, it appears that functional foods and nutraceuticals are barely distinguishable since nutraceuticals, a word coined from two words, nutrition and pharmaceutical in 1989 by Stephen DeFelice, has been defined as a food or any part of a food that provides medical or health benefits, including the prevention and/or treatment of a disease [17]. In fact, in certain countries, both functional foods and nutraceuticals are used interchangeably [18]. Both functional foods and nutraceuticals contain bioactive components that contribute to improving health and reducing the risk of disease in humans. Because the Moringa plant contains a suite of chemicals and has a reservoir of bioactive secondary metabolites, which are mainly responsible for its medicinal value, the plant is regarded as a functional crop with nutraceutical properties. For example, historically in India, Maurian warriors were fed with the leaf extract on the front lines to provide additional vitality and reduce the pressure and torment during war [7]. These exceptional characteristics of the extract have been attributed to the presence of bioactive compounds, such as chlorogenic acid, ellagic acid, ferulic acid, gallic acid, quercetin, and kaempferol, with several nutritional and pharmaceutical properties [21]. More recently, an in vivo study by Cuellar-Nuñez et al. [22] showed that the leaf decreased the activity of harmful fecal enzymes such as β-glucosidase, β-glucuronidase, tryptophanase, and urease and showed potential for preventing colon cancer. This chapter presents a comprehensive review of the role of Moringa in nutraceuticals and functional foods. This will include its nutritional and traditional uses, as well as the antiasthmatic, antibacterial, anticancer, antidiabetic, antifungal, anti-inflammatory, antioxidant, antiviral, hepatoprotective, and neuroprotective properties of the seed, leaf, and flower.

6.2. Nutritional value of Moringa oleifera

All the parts of Moringa, including the flower, leaf, and seed, are rich sources of nutrients, but variations in composition have been observed among these parts (Tables 6.1–6.3) as well as among different varieties of the plant. For example, Jongrungruangchok et al. [31] reported significant variations in the fiber (16%–24%) and protein contents (approx. 19%–29%) of leaves of Moringa grown in 11 different provinces in Thailand. The advocacy for the use of Moringa in various food applications has been associated with the exceptionally high protein content in the leaf and seeds (Table 6.1). Moringa flower, leaf, and seed also contain appreciable quantities of amino acids (Table 6.2). Generally, the total amino acids concentrations were higher in the leaves than in the flowers and seeds [46], including two to three times higher essential amino acids. Protein hydrolysates from Moringa seeds, and perhaps the flowers and leaves, may have the potential for industrial applications as functional ingredients and nutraceuticals due to their antioxidant activity and ability to inhibit angiotensin-converting enzyme [52]. Only a few studies have been documented on the amino acid profile of Moringa flower suggesting the need for future studies on this part of the plant [46]. Besides the protein content, the flowers, leaves, and seeds are also excellent sources of essential oils that contain high levels of essential fatty acids (Table 6.3). Compared to the leaf and flower, Moringa seed is richer in oil, which may vary between 2.60% and 45.60% [30,32,41]. Oleic acid is the most abundant fatty acid in the flower (24.09%–29.00%) [54,55] and seeds (66.50%–85.52%) [35,41,57], while linolenic acid (33.00%–54.27%) is the most abundant in the leaves [33,51,54,56]. The leaves generally contain low amounts of saturated fatty acids, but high levels of mono- and polyunsaturated fatty acids [41,58,59], which can be used to improve the functionality and health benefits of Moringa-based products [41]. Further to this are the possibilities of utilizing the oil in reducing rancidity in fatty foods due to its high antioxidant capacity [60], which is associated with the high levels of tocopherol [41].

Table 6.1

Proximate composition (g/100g) of Moringa oleifera flower, leaf, and seed.

Moringa part | Ash | ∗CHO | Fat | Fiber | Moisture | Protein | References
Flower | 1.89–8.17 | 46.45–56.89 | 2.95–10.27 | 2.17–17.27 | 8.97–21.82 | 3.07–30.08 | [23–29]
Leaf | 1.96–15.22 | 8.10–63.11 | 0.17–19.00 | 3.22–33.20 | 3.21–17.75 | 14.26–38.98 | [22–24,30–38]
Seed | 2.37–9.97 | 0.10–57.77 | 2.60–45.60 | 0.10–21.00 | 4.10–10.50 | 9.98–43.71 | [7,23,24,30,32,35,36,38–45]

∗
CHO: Carbohydrate.

Table 6.2

Amino acid composition (g/100g) of Moringa oleifera flower, leaf, and seed.

Amino acid | Flower | Leaf | Seed
Ala | 8.10 | 1.05–9.79 | 4.41–7.07
Arg | 20.10 | 1.78–14.33 | 5.85–13.19
Asp | 12.30 | 1.43–9.22 | 6.98
Cys | | 0.01–5.60 | 0.67–3.42
Gta | 17.00 | 1.03–15.10 | 15.20
Gly | | 1.36–10.57 | 4.34–12.95
Ile | 5.20 | 1.13–5.03 | 2.92–3.54
Leu | 8.70 | 1.96–9.70 | 5.64–7.17
Lys | 4.60 | 1.41–5.82 | 1.29–3.30
Met | 1.00 | 0.29–1.98 | 1.44–1.93
Phe | 3.80 | 1.55–6.18 | 3.67–5.35
Pro | 6.60 | 1.20–5.70 | 4.37–6.13
Ser | 7.50 | 1.09–5.75 | 3.19–3.71
Thr | 5.40 | 0.75–5.28 | 3.30–4.09
Val | 6.40 | 1.41–6.65 | 3.68–7.08
Tyr | 0.40 | 0.97–3.87 | 1.71–2.27
References | [46] | [33,46–51] | [46,50,52,53]

Ala: Alanine; Arg: Arginine; Asp: Aspartic acid; Cys; Cysteine; Gta: Glutamic acid; Gly: Glycine; Ile: Isoleucine; Leu: Leucine; Lys: Lysine; Met: Methionine; Phe: Phenylalanine; Pro: Proline; Ser: Serine; Thr: Threonine; Tyr: Tyrosine; Val: Valine.

Differences have been observed in the mineral composition of Moringa flower, leaf, and seeds (Table 6.4), and this may be associated with the growing conditions and location, variety, storage conditions, and the stage of maturity at harvesting. For instance, the calcium and iron contents of matured Moringa leaves were three times and one and a half times higher than in young leaves, respectively [63]. The leaf of Moringa is generally rich in calcium and potassium [24,61,62], while the leaf and seeds are good sources of iron and zinc [24,29]. Since the fruit and leaves of Moringa are usually consumed more frequently, they can be potential sources of iron in the diet, enhancing the formation of hemoglobin and myoglobin and preventing anemia [64]. The rich levels of these minerals in the flower, leaf, and seed are indicative of their potential for food fortification.

Table 6.3

Selected fatty acid composition (g/100g) of Moringa oleifera flower, leaf, and seed.

Morina part | Myristic | Palmitic | Stearic | Arachidic | Oleic | Linoleic | Linolenic | References
Flower | 0.16–1.45 | 24.25–33.60 | 5.54–5.87 | 0.43–1.23 | 24.09–29.00 | 15.48–18.60 | 10.60–18.82 | [54,55]
Leaf | 0.22–3.66 | 11.79–54.30∗ | 2.13–6.08 | 1.06–1.61 | 3.30–4.93 | 4.23–13.35 | 33.00–54.27 | [33,51,54,56]
Seed | 0.07–1.93 | 6.55–24.33 | 0.72–9.15 | 0.47–3.78 | 66.50–85.52 | 0.62–16.91 | 0.07–30.42 | [35,41,57]

∗Value reported as percentage total fat.

Table 6.4

Mineral composition (mg/100g) of Moringa oleifera flower, leaf, and seed.

Moringa part | Ca | Mg | K | Na | Fe | Zn | Cu | References
Flower | 2.32–389.90 | 36.17–154.32 | 3.02–3607.90 | 77.41–813.90 | 33.49–98.11 | 6.53–14.19 | 0.18–4.04 | [24,25,28,29]
Leaf | 1255.30–4192.00 | 43.98–1555.00 | 1845.00–2044.00 | 8.13–94.21 | 19.37–106.00 | 1.00–7.25 | 0.55–1.03 | [24,61,62]
Seed | 111.50–216.60 | 47.30–104.60 | 47.10–581.80 | 15.90–36.17 | 12.30–132.30 | 6.77–28.03 | 0.18–0.46 | [24,29]

6.3. Traditional food and potential uses of Moringa oleifera

Moringa is an exceptional plant because of its vast medicinal properties [65]. It is gaining popularity in both developing and developed countries [66]. The plant has gained importance due to its ability to thrive well in different climatic regions (dry and humid) [66]. Moringa leaves, seeds, and flowers contribute significantly to human nutrition due to their nutritional composition and have also been employed in alternative medicine [5,67,68]. Traditionally, the Moringa plant is used as an antispasmodic, diuretic, expectorant, and stimulant [69]. Table 6.5 summarizes some food uses of the plant, especially in food enrichment and fortification for improving the nutritional value of traditional foods. Moringa flower, leaf, and seed powder have been used in the fortification of different food products such as amala (stiff dough), bread, biscuits, cheese, ogi (maize gruel), soups, yoghurt, and weaning foods [5,67,86].

Fresh leaves are eaten as a snack and integrated into salads and vegetable soups [68,70]. Powdered leaves are included in the diets of pregnant and lactating women (to increase milk production) and children (to combat malnutrition) [67]. They are also used as condiments, spices, teas, infusions, and flavorings [68]. The low fat content of Moringa leaves makes them suitable for diets focused on managing obesity [87] and in value-added food products [88]. The use of the Moringa leaf, although it has potential for use in food applications, has an intense green color that limits its use in foods. Consumer acceptability of products enriched with the leaves has been reported to be unacceptable [73,74]. Efforts have been made to decolorize the leaf to enhance its use in food applications [89]. However, the use of decolorized leaves is yet to be documented in the literature, suggesting a research gap for future studies. The Moringa leaf extract can be a great source of natural antioxidants in cooking oils.

In India, for instance, the seeds are processed into a drink called Zija [80]. Seeds are eaten raw or roasted like groundnut. Ground seed is added to water as a coagulant for water purification with the added benefit of antifungal and antibacterial compounds [79,90]. The seeds are also believed to be a low-cost biosorbent with the ability to remove metals from wastewater [87]. They have other applications in animal forage, biogas, domestic cleaning agents, and green manure [66,90]. The seed oil is used as a lubricant, and in perfumes, hairdressing, and cooking [67]. The use of seed cake and leaves as a fertilizer has also been reported [67]. Seeds are also believed to be effective in regulating blood pressure and are also employed in the treatment of diabetes, fever, stomachache, malaria, and sexual dysfunction. The seed extract incorporated during cottage cheese production was found to significantly increase cheese yield [91]. Moringa flower is reportedly used in enriching weaning foods [85,86] and bread [81]. It is envisaged that Moringa leaf, seed, and flower will be globally used in food formulations as fortificants to improve human nutrition by serving as a cheap source of essential nutrients.

6.4. Antimicrobial properties

Plants generally produce a wide range of secondary metabolites which not only serve to protect them from potential environmental threats [92] but also have different pharmacological activities [65]. The different parts of the miracle tree are known to be effective in inhibiting the growth of different microorganisms.

Table 6.5

Some food uses of Moringa oleifera.

Moringa part | Traditional uses | References
Leaf | Fresh | Fresh leaves are eaten as a snack, and in salads and vegetable soups | [68,70,71]
Leaf juice is used to relieve hiccups, and cooked leaves are for treating influenza and catarrhal | [69]

Dry powdered

Included in diets for pregnant and lactating women (to increase milk production)

Given to children to combat malnutrition

[67]

Condiments, tea, infusions | [68]
Fortification/enrichment | Bread | [72]
Amala | [73,74]
Biscuit/cookies | [75–78]
Seed | Seeds are eaten raw and roasted like groundnut | [79]
Drink (Zija) | [80]
Fortification/enrichment | Bread | [81]
Biscuit/cookies | [82]
Seed oil is used in cooking | [67]
| Seed extract used as thickener in yoghurt | [83]
Flower | Fresh and dried flowers are used for tea | [84]
The flowers can be eaten raw with salads, blanched or fried | [84,85]
Weaning food formulation | [85,86]
Bread | [81]

Phytochemical screening of Moringa leaf confirmed the presence of alkaloids, phenols, saponins, and tannins in its water, acetone, and ethanol extracts [93]. Nwosu and Okafor [94] observed that Moringa leaf extract was able to inhibit the growth of some fungal species. Dahot [92] found different fractions of Moringa leaf protein to be effective in preventing metabolic activities of certain bacteria and fungi species and thus inhibiting their growth. The antibacterial activity of these protein fractions was assessed against Escherichia coli, Klebsiella aerogenes, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus subtilis. All the fractions had a strong inhibitory capacity against E. coli, S. aureus, and B. subtilis. Also, the leaf extract used in puffer-fish-skin-gelatin film for gouda cheese packaging was found to efficiently inhibit microbial growth and delay lipid oxidation [95].

Moringa seed ethanolic extracts were found to be rich in tannins, alkaloids, flavonoids, and cardiac glycosides, but lacking in saponins, proteins, and amino acids [65]. A Moringa seed aqueous extract and the juice from its fresh leaf reportedly inhibited the growth of Pseudomonas aeruginosa and S. aureus [69]. Also, seed extract was found to inhibit Bacillus cereus, Candida albicans, Streptococcus faecalis, B. subtilis, Staphylococcus epidermidis, S. aureus, P. aeruginosa, E. coli, and Aspergillus niger [96]. However, ethanolic extracts were found to be more effective than water and acetone extracts, in inhibiting the bacteria Salmonella typhi which causes typhoid fever. The inhibitory capacity of all the extracts was said to be similar to that of commercially available antibiotics such as chloramphenicol, ciprofloxacin, and cotrimoxazole [93]. Increasing the temperature up to 100°C did not affect the inhibitory activity of the extracts, but increased pH (8–10) did reduce activity [93]. Jabeen et al. [97], in their study on the antimicrobial activity of seed extract against some bacterial (B. subtilis, E coli, Pasteurella multocida, and S. aureus) and fungal (Fusarium solani and Rhizopus solani) strains, found that the zones of growth inhibition showed greater sensitivity against the bacterial strains than the fungal strains. Minimum inhibitory concentrations showed P. multocida and B. subtilis to have a higher sensitivity to the extract than other organisms. Fungal growth inhibition was through paralysis of their hyphae and apical branching. The effectiveness of the extracts was however limited by cations such as Na+, K+, Mg2+, and Ca2+, and the greatest activity was between 4 and 37°C and at pH 7 [97].

Moringa oil was found to prevent the proliferation of Listeria monocytogenes on fresh hard cheese (Camembert, Cheddar, Mozzarella, and Parmesan) [98]. This action was proposed to be due to the inhibition of the Embden–Meyerhof–Parnas pathway and the loss of intracellular DNA of the bacteria. Moringa oil has also been used in the biocontrol of L. monocytogenes and S. aureus on cheese [99]. Few studies have been done on the antimicrobial properties of Moringa flower, suggesting the need for future research.

6.5. Anti-inflammatory properties

Living organisms, as a means of defense, produce inflammatory responses resulting from the expression of nuclear factor kappa B, cytokines, and nitric oxide [100]. Inflammation is the body's protective response to tissue injury resulting from harmful chemicals, microbes, or physical trauma [65]. It is a route to the healing process of the tissue. Commercially available nonsteroidal anti-inflammatory drugs have been found to pose different health risks and alternative drugs are being sought [100]. The anti-inflammatory activity of Moringa has been previously reported in the literature [87,101,102].

Anti-inflammatory activity of Moringa has been associated with its nitriles, glycosidic glucosinolates, isothiocyanates, carbamates, thiocarbamates, quercetin, quercetin-3-O-glucoside, kaempferol glucoside, 3,5-dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4–1,9-octadecenamide, crypto chlorogenic acid, aurantiamide acetate, 1,3-dibenzyl urea, and 4(α-L-rhamnosyloxy)-benzyl glucosinolate content [87].

Alhakmani et al. [65] studied the in vitro anti-inflammatory activity of Moringa flower extract at different concentrations (100–500μg/mL) compared with diclofenac sodium in thermally induced protein denaturation in egg albumin. The inhibition of inflammation resulting from protein denaturation was found to increase with increasing Moringa flower extract concentration.

Hot water infusions of Moringa leaves, roots, seeds, or stalks have also been reported to have anti-inflammatory activities on induced edema in rats [69].

The aqueous leaf extract has been found to downregulate a pro-inflammatory transcription factor in cultured human pancreatic cancer cells [88]. The presence of isothiocyanates (1.66%) and total polyphenols (3.82%) in Moringa extracts has been associated with a significant reduction in gene expression and production of inflammatory markers in RAW macrophages, an indication of its suggested ability to reduce mild inflammation associated with chronic diseases [88,103].

Ethanolic extract of Moringa leaves has been found to inhibit chemotactic oxidation of polymorphonuclear leukocytes and keratinocytes [87]. Extracts (ethanol-water mixture) of Moringa seeds and leaf reduced bowel inflammation and enhanced cellular and humoral immunity in rodents with weakened immunity, respectively. Also, hydroalcoholic extracts from flowers were found to decrease the movement of incendiary mediators and pro-inflammatory cytokines in macrophage cells [104].

6.6. Antioxidant properties

Oxidative stress can be tackled effectively with an antioxidant; antioxidants generally will sacrifice themselves to be oxidized. One antioxidant molecule would normally react with a single free radical, and the used up antioxidant would need to be replenished, either through endogenous or supplementation means [105]. Interestingly, extracts from Moringa plants are reported to be rich in antioxidant compounds [106,106]. In the body, cells could be attacked by free radicals, and these free radicals can be prevented from performing their destructive role when antioxidants are present in adequate amounts [107]. Oxidative stress is one of the leading causes of cancer [108], and the presence of antioxidants can prevent cancer by disrupting free radical formation and reducing oxidative stress.

The biological activities, including antioxidant properties of Moringa could be due to the presence of natural active ingredients (phytochemicals), such as flavonoids and some phenolic compounds that may be available in the leaf extract [109]. Natural antioxidants and preservatives, utilized in the form of plant extracts in the food industry, are receiving great attention because of the health benefits to be derived by humans that consume food products with these active ingredients, and the ability to prolong a product's stability and availability. Antioxidants normally prevent oxidative stress, which would otherwise cause degenerative diseases due to an overabundance of reactive oxygen species. The active components of the plant detoxify the reactive intermediates or rectify any damage that may be formed [110]. Moringa plants are not only good sources of nutrients, but also are equally useful worldwide for the treatment of various ailments ranging from rheumatism, ascites, infections, hiccough, influenza to an internal abscess [16,69] and in other non-food applications [111–113]. Additionally, the leaf extracts have been reportedly used to reduce hyperglycemia and dyslipidemia [21] and defects in mice resulting from cyclophosphamide-induced micronucleus formation and DNA damage [114]. Hepatic glutathione could be restored with the use of an aqueous extract of Moringa leaves [115]. Other authors found that the tender pods of Moringa inhibited azoxymethane-induced colon carcinogenesis [116] and prevented the formation of erythrocyte micronucleus in mice injected with 7,12-dimethylbenz(a)anthracene [117]. The leaf extract also showed inhibitory action against oxidative DNA damage in human keratin-forming tumor cells [118] and significantly inhibited the growth of pancreatic cancer cells as well as enhanced the efficacy of chemotherapy in human pancreatic cancer cells [119].

As a result of these, food antioxidants are vital in human nutrition, as they decrease the oxidative damage to food components such as the lipids, proteins, and nucleic acids that induce free radical formation [120]. Consumption of fruits and vegetables is reported to be linked with a lowered risk of diseases in humans. Young leaves, flowers, and some other parts of Moringa are consumed worldwide, most especially in Africa and India, Pakistan, Philippines, and Hawaii [16,121]. Their consumption is believed to result in a lower risk of diseases as a result of the beneficial active ingredients in the plants.

The oil from the Moringa seed compares favorably with olive oil in terms of nutritional profile [41] and contains appreciable amounts of antioxidants indicating their potential in preventing rancidity in food systems. Some plant extracts are well known to prevent free radical formation and can also block the propagation of autoxidation in different foods [122]. The extracts have possible preservative properties, i.e., numerous bioactive compounds that have the potential to destroy microorganisms by disrupting the cytoplasmic membrane of the cells [123,124]. Future studies on the use of natural plant extracts in preventing unfavorable changes due to the formation of rancid flavor in fatty foods need to be investigated. This process needs to be prevented using a potent preservative that would be less toxic or harmful than artificial ones. To overcome these challenges, natural additives, mostly from plant sources, are being used [122,125]. For example, Nandave et al. [126] reported that Moringa leaf extract significantly reduced lipid peroxidation in the myocardial tissue of Wistar albino rats, indicating the cardioprotective effect and antioxidant activity of the extract. The antioxidant potentials of Moringa leaf extract fractionated into five parts including aqueous, crude, diethyl ether, phenolic, and polyphenolic fraction using different antioxidant test models were equally explored by Verma et al. [127]. The antioxidant activities of the five fractions were found to be in the order of aqueous<nonphenolic<diethyl ether<crude extract<polyphenolic fraction. The polyphenolic fraction with the highest antioxidant activity, at a concentration of about 10μg/mL, also showed strong protection to DNA damage, further confirming its strong antioxidant properties. In another study to ascertain the efficacy of Moringa parts, Sreelatha and Padma [118] investigated the antioxidant and cytotoxicity activities of mature and tender leaves of Moringa extracts. Matured leaves showed higher levels of phenolic compounds (almost double) than the tender ones, suggesting higher antioxidant activity in the former [118]. For example, the levels of quercetin (795μg/mL) and kaempferol (216μg/mL) in the matured leaves were higher than in the tender leaves (quercetin: 436μg/mL; kaempferol: 115μg/mL). The higher level of phenolic compounds in the matured leaf may explain its higher antioxidant activity (approx. 85%) compared to the tender leaves extract which showed approximately 64% activity [118]. Both leaf extracts also showed potential to prevent H_2O_2-induced DNA damage in KB cells (subline of the ubiquitous keratin-forming tumor cell line HeLa), but the matured leaf extracts displayed better or higher cell viability than the tender ones.

6.7. Anticancer properties

All parts of the Moringa plant have phytochemicals that are useful medicinally. The World Health Organization reported cancer as the second leading cause of death globally, with estimated deaths of around 8.8 million [128]. According to the report, the occurrence and death from cancer are even expected to increase by as much as 70% as the years roll by. The rate of deaths from cancer is higher in less developed countries, possibly due to increased pollution levels, increase in life expectancy, and insufficient healthcare facilities coupled with the cost of purchasing anticancer drugs, among others. The best way to tackle this menace is to look inward by developing anticancer drugs from traditional plants like Moringa, which may be far less toxic and would be more affordable for the populace of the less developed countries than existing pharmaceuticals. A majority (over 60%) of existing anticancer drugs originated from plants and natural sources such as marine organisms and microorganisms [129]. For example, camptothecin is an anticancer drug that is obtained from Camptotheca acuminata plant extract [130]. In comparison with conventional drugs, phytochemicals from plants have been reported to have very little or no adverse effects, with very low toxicity, leading to more research work on medicinal plants [131]. One of the numerous ailments supposedly treated with Moringa is cancer, which has very high mortality worldwide. Cancer may result from the diet, lifestyle, and environment, among other factors [132,133]. The relatively high amounts of phytochemicals in Moringa plant [134] and the low toxicity of the extract [135] suggest the plant is a good source of anticancer drugs. Researchers have nursed a great interest in exploring the numerous compounds present in the various parts of the Moringa tree, most especially for the potential treatment of various ailments, including cancer. These research activities, demonstrated through in vitro and in vivo testing, showed the medicinal potential of various extracts from the leaves, roots, shoots, seeds, flowers, and bark of the Moringa plant. However, the majority of the research on the anticancer activity focused on the use of leaf extract. The anticancer activity of the leaf has been associated with a high level of polyphenols and flavonoids [136]. The anticancer potentials of plant extracts could be measured by in vitro assay on cancer cell lines by inspecting their impact on the growth and proliferation of cancerous cells. To build up the evidence base for any promising anticancer activity, the particular pathway of disruption in specific cancer cell lines needs to be elucidated. Once a feasible mechanism has been established, researchers will expand their exploration of the anticancer activity of the extract in vivo, using mouse or rat models. These represent a better model because they include systemic metabolism effects. Johnson [131] reported that the low toxicity of phytochemicals derived from plant extracts justifies more research work on medicinal plants. In a study by Parvathy and Umamaheshwari [137] on Moringa leaf extract and its effects on the human B-lymphocyte plasmacytoma-U266B1 cell line, the methanolic extract exhibited the highest cytotoxic activity against U266B1 cells when compared with the extracts of ethanol, ethyl acetate, and chloroform. This suggests that leaf extracted with methanol showed very high anticancer activity. Nair and Varalakshmi [138] equally reported the anticancer and cytotoxic potential of extracts of the plant carried out with hot water, methanol, and hexane. The authors found that extracts obtained with a mixture of methanol and hexane led to a decrease in HeLa cell viability (IC_50: 70μg/mL) with increasing concentrations [138]. Charoensin [139] also studied the antioxidant and anticancer activities of methanol and dichloromethane Moringa leaf extracts and reported higher antioxidant activity in the methanolic extract. However, the dichloromethane leaf extract (IC_50: 112–133μg/mL) showed higher anticancer activity than the methanol extract (250μg/ mL). The anticancer effect of cold water, hot water, and 80% ethanolic leaf extract was also reported by Al-Asmari et al. [140] using primary leukemia cells harvested from 15 patients with acute myeloid leukemia (AML) and 10 patients with acute lymphoblastic leukemia (ALL). The anticancer activity of the extracts was in the order ethanol>hot water>cold water in both AML and ALL cells.

Other parts of the tree such as seed, flower, and bark of the Moringa tree have also been shown to possess anticancer properties. Al-Asmari et al. [141], for example, examined the effects of ethanol extracts of the leaf, seed, flower, and bark of Moringa on ileocecal adenocarcinoma (HCT-8) and human breast adenocarcinoma (MDA-MB-231) cell lines. Phenotypic evaluation showed that 500μg/mL of the extracts caused motility reduction (reducing the chance of cancer occurring) of about 50% for the bark extract, but no detectable motility reduction for other extracts.

Soler-Rivas et al. [121] also studied the anti-inflammatory and antitumor properties of crude, dried, and green seed ethanolic extracts of Moringa. The extracts were concentrated and reconstituted in hexane, ethyl acetate, butanol, and water, and the anti-inflammatory activities of the extracts were tested using an in vivo method (mice with carrageenan-induced inflammation in the hind paw). To ascertain the efficacy of these tree parts on antitumor activity, the ability of the extracts to prevent the formation of Epstein–Barr virus-early antigen induced by 12-O tetradecanoylphorbol-13-acetate, which could aggravate tumor formation, was studied. The crude ethanol extracts prevented its formation at a dose of 100μg/mL. Furthermore, the anticancer potential of the methanolic extract of the seed using cancer-induced animal, Hep-2 (HeLa contaminant carcinoma), 502713 HT-29 (colon cancer), and IMR-32 (neuroblastoma) cell lines was reported by Rajesh et al. [142]. The extract showed 80% inhibition in A549 cells (human lung adenocarcinoma) indicating the antiproliferative activity of the extract), 95% inhibition in 502713 HT-19 cells, and 93% inhibition in IMR-32 cells, but it did not affect Hep-2 cells. Overall, the Moringa seed ethanol extract displayed very high anti-inflammatory activity and could be an antitumor product.

6.8. Antidiabetic properties

The antidiabetic properties of Moringa have also been widely studied in the literature [143–147]. Diabetes is a sickness defined by chronic hyperglycemia resulting from abnormalities in insulin secretion, insulin action, or a combination of the two, and accompanied by serious consequences. There are growing global concerns on the impact of diabetes mellitus on human health, since the disease has several associated risks and could lead to untimely death in severe cases. Over 422 million people worldwide have developed the deadly disease, diabetes mellitus [147]. Almost all the various parts of the Moringa plant have antidiabetic activity including the flower [148], leaf [147–150], seed [148,151], and roots [147,148,152,153].

Adejoh et al. [148] studied the in vitro and in vivo antidiabetic potentials of ethanolic extracts of Moringa flowers, leaf, roots, seed, and stem. The in vivo study used diabetic rats, induced with 150mg alloxan intraperitoneal injection per kg body weight, and found that the seed and leaf extracts showed greater glucose-lowering compared to other parts (flower, stem, and roots) and hence higher antidiabetic properties [148]. In another study, the efficacy of Moringa leaf extract (Moringinine), compared with quercetin and chlorogenic acid, was tested against alloxan-induced diabetic rats, and the result showed that their effectiveness in lowering blood glucose was in the order quercetin>chlorogenic acid>Moringinine [143]. However, in a different study, Moringa extracts
(leaf and roots) generally showed higher antihypoglycemic activity compared to the acarbose reference, though the roots displayed higher activities than the leaf [147]. Ali et al. [143] further reported that Moringinine normalized the elevated serum levels of glucose, triacylglycerol, and total cholesterol. Furthermore, the extract reportedly restored the normal histological structure of the pancreas in diabetic rats [143].

Earlier studies on the antihyperglycemic activity of Moringa leaf extracts using human subjects also showed substantial reductions in serum glucose concentrations during the test period [154,155]. The antidiabetic activities of the different parts of Moringa plant are associated with the presence of various phytochemicals and antioxidants, such as quercetin and kaempferol. Other possible mechanisms for the antidiabetic activity of the Moringa plant, as suggested by earlier researchers, include the release of insulin by inhibiting the adenosine triphosphate–sensitive potassium channels in the membrane of the residual beta-cells, just like sulfonylureas and meglitinides, and enhanced stimulation of insulin to arouse glucose uptake and utilization by adipose, liver, muscle, and skeletal tissues [148,156]. It may also involve the quick inhibition of α-amylase and α-glucosidase enzymes, which are involved in the breakdown of dietary starch to glucose [157]. These enzymes are presumably inhibited by the presence of bioactive compounds such as quercetin, kaempferol, and chlorogenic acid. Habtemariam [158], for example, found rutin and neochlorogenic acid as bioactive components responsible for the antidiabetic properties of Moringa stenopetala leaf extract.

6.9. Antiasthmatic properties

Moringa species contain alkaloids, tannins, phenolic acids, saponins, flavonoids, terpenes, steroids, and glucosinolates, which are regarded as phytochemicals and are major materials for pharmacological purposes [159]. Asthma is characterized by chronic respiratory disorder as a result of inflammation of the bronchi [160]. Studies have shown that Moringa leaf powder and seed extract have antiasthmatic properties [160–163]. Mahajan et al. [163] stated that the antioxidant, anti-inflammatory activities and histopathological examination showed that Moringa extract has a protective effect on the immune-mediated inflammatory responses in rat. According to Goyal et al. [162], Moringa leaf extract inhibits the release of the inflammatory mediator histamine and affects the immune response and inflammation in bronchial asthma and inflammatory cells. The antiasthmatic activity of alcoholic extract of Moringa seed on different experimental animals with bronchial asthma was reported by Mehta and Agarwal [160]. It was observed that Moringa had a bronchodilating effect on the animals which was comparable to ketotifen fumarate. According to the authors, the ethanolic extract of Moringa increased the preconvulsion time in animals induced with acetylcholine aerosol and histamine, thereby reducing the high monocyte and eosinophil counts in the bronchoalveolar lavage fluid of pigs [160,162]. Suresh et al. [164] similarly observed that the methanolic extract of Moringa leaf can prevent bronchoconstriction, airway inflammation, and asthma. Improvement in these parameters following administration of Moringa extract indicated that the seed kernel could be useful in treating patients with bronchial asthma [160,161]. The ethanolic extract of Moringa seeds tested on guinea pigs induced with ovalbumin airway inflammation was reported to show a significant increase in respiratory parameters and reduction in interleukins in bronchoalveolar lavage [165]. The effectiveness of the Moringa seed kernels in controlling asthma has been associated with its antimicrobial/anti-inflammatory activities, mast cell stabilization, bronchodilator action, and inhibition of immediate hypersensitivity reaction [160,162].

6.10. Hepatoprotective properties

The hepatoprotective activities of ethanolic extracts of Moringa leaf have been studied in rats with liver damage induced by antitubercular drugs such as pyrazinamide, isoniazid, and rifampicin [166]. It was discovered that the oral administration of Moringa extract enhanced the recovery from hepatic damage as demonstrated by the histopathological examination of liver tissues. Improvement was observed in the concentrations of alkaline phosphatase, bilirubin, glutamic oxaloacetic transaminase, and glutamic pyruvic transaminase in the serum [166]. Likewise, Mousa et al. [167] studied the protective activities of an ethanolic extract against thioacetamide toxicity in rats and observed a decrease in hepatic glutathione level, reduced pathological alterations, reduced serum activities of enzymes (aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase), and lipid peroxidation in the hepatic tissue. The ethanolic extract of Moringa leaves had both ameliorating and protecting potentials against the thioacetamide-induced liver damage through the regulation of antiapoptotic, antioxidant, and inflammatory biomarkers [167]. The hepatoprotective properties of ethanolic extracts of Moringa leaves, roots, and flowers in rats with drug-induced hepatotoxicity from aluminum chloride, acetaminophen, carbon tetrachloride, diclofenac sodium, Salmonella typhimurium, and other toxicants have been studied [168–175]. Moringa ethanolic extract reduced the deleterious effects of these drugs on the liver enzymes and improved liver function. Moringa leaf prevented hepatic steatosis by affecting gene expression related to hepatic lipid synthesis, which resulted in lower levels of cholesterol, triglycerides, and reduced inflammation in the liver [176]. Previous studies observed that the hepatoprotective properties of Moringa leaves have an antioxidant-mediated mechanism derived from the bioactive compounds present in the plant [177,178]. Hepatoprotective activities of Moringa have been linked to kaempferol and quercetin, which are flavonoids, and antioxidants such as vitamin A and ascorbic acid [175,179].

6.11. Neuroprotective properties

Neuroprotection relates to the ability of a chemical substance to preserve neuronal function or structure. Therefore, neuroprotective agents protect the brain from neuronal injury or neurodegenerative diseases [180]. Moringa serves as a potent neuroprotectant due to the presence of antioxidants, which reduce the reactive oxygen species produced by reperfusion and lipid peroxidation, thereby protecting the brain from cerebral ischemia caused by the obstruction of blood flow to the brain [84,181,182]. In a study carried out by Hashim et al. [183], it was reported that Moringa leaf extract has neuroprotective actions within the brain, with the potential to promote learning and cognitive functions and reduce neuroinflammation. According to Sutalangka et al. [184], Moringa leaf extract has been used in treating dementia due to its efficacy in decreasing acetylcholine esterase activity resulting in improved cholinergic function and memory. Methanolic extract of Moringa leaves has been shown to decrease the toxic effects of vanadium-induced cytotoxicity in rat due to its antioxidant properties. The study conducted by Igado et al. [185] showed that a methanolic extract of Moringa leaves reduced the neurobehavioral deficit induced by vanadium and had neuroprotective activity against microglia activation, neuronal loss, astrocytosis, and demyelination. The neuroprotective effect of the Moringa leaf extract was investigated using aluminum-induced cortical degeneration in rat [186]. The study observed that the administration of Moringa extract showed reduced degenerative features, increased neuron-specific enolase, and decreased glial fibrillary acidic protein expression [186]. The neuroprotective properties of the Moringa leaf extract were due to the presence of polyphenolic compounds with strong anti-inflammatory and antioxidant activities against neurodegeneration [185,187]. Giacoppo et al. [188] and Igado and Olopade [189] also observed the presence of glucosinolates, their products, and isothiocyanates in Moringa plant, which contributed to the neuroprotective effect against neurodegenerative diseases.

6.12. Conclusion and future directions

This chapter reviewed the roles of the various parts of M. oleifera in nutraceuticals and functional foods. The parts of the plant (leaf, seed, and flower) are known to exhibit anticancer, antidiabetic, antioxidant, anti-inflammatory, and chemopreventive activities and as such have potential for food production applications. The bulk of the research on Moringa focused mainly on the leaf, with less information available on the seed and flower, despite the excellent nutritional profile of the seed and flower. Furthermore, the use of these parts (flower, leaf, and seed) within the food and nutraceutical production industries has not been fully exploited. Hence, more studies on the seeds and flowers as well as increased practical applications in food systems, to validate the claims in the literature, are required. The use of gas chromatographic techniques to elucidate the metabolomic profile of the flower, leaf, and seeds as well as in products enriched with these plant parts are equally important to fully understand the active health components in the Moringa plant.

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Chapter 7: Galangal—Roles in nutraceuticals and functional foods

Augustine Amalraj, Karthik Varma A.C., Shintu Jude, and Sasikumar Kuttappan
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Abstract

This chapter discusses briefly about the galangal, including greater galangal (Alpinia galanga (L.) Willd) and lesser galangal (Alpinia officinarum Hance), which are broadly used in several foods and traditional medicines in Southern Asia and Indonesia because of the presence of numerous bioactive components particularly galangin that have various medicinal and pharmacological properties such as antiinflammatory, antioxidant, anticancer, cardiovascular, liver health, neuroprotective, antiviral, antimicrobial, reproductive, and immune-stimulating activities. Galangal are widely used as flavoring agent in foods, sausages, spicey condiments, and herbal tea. Therefore, galangal can be the hopeful spices to develop a diversity of innovative functional foods and nutraceutical formulations that could assist satisfy the necessity of the human health and wellness due to the various biological activities.

Keywords

Alpinia galanga; Alpinia officinarum; Biological activities; Functional foods; Galangal; Nutraceuticals

7.1. Galangal and its phytochemistry

Galangal terms are used to designate Alpinia galanga (L.) Willd and its closely associated species Alpinia officinarum Hance, which are members of Zingiberaceae family [1]. A. galanga generally known as galangal or greater galangal is a perennial plant, native to Indonesia, however cultivated across Asia, with Indonesia serving as the primary producer and supplier. A. officinarum is recognized as lesser galangal that has the same properties and usages of A. galanga, even though this is the most significant in relations of culinary practices [2]. The chemical compositions of A. galanga and A. officinarum, on the other hand, differ. A. galanga contains a wide range of phenolic components and essential oils, according to phytochemical study. Meanwhile, the A. officinarum rhizome is high in flavonoids, diarylheptanoids, and essential oils. Diterpenes, glycosides, phenylpropanoids, and sesquiterpenes are some of the additional components that can be isolated from A. galanga and A. officinarum [3].

A. galanga has a lot of phenolic substances like phenylpropanoids (Fig. 7.1). One of the major phenylpropanoids [4], 1′S-1′-acetoxychavicol acetate, is recognized as the source of A. galanga's unique pungent and fragrance component. Flavonoids such as galangin (3,5,7-trihydroxyflavone) and galangoisoflavonoid have also been found in the rhizomes of A. galanga [5,6]. Nine flavanols, galangin-3-methyl ether, galangin, kaempferide, kaempferol, quercetin-3-methyl ether, quercetin, apigenin, isorhamnetin, and rhamnocitrin; two flavanonols, pinobanksin and pinocembrin; and epicatechin are the major constituents of A. officinarum [3]. Galangin is the major ingredient in A. officinarum that is found in high amounts.

Despite the fact that the quantities of active constituents in these two plants differed significantly, both A. galanga and A. officinarum were well-known traditional Chinese medicines and were widely used as a remedy for gastrointestinal diseases such as dyspepsia, gastrofrigid, stomach ache, and vomiting. A. galanga and A. officinarum have been found to have a variety of biological actions, including antioxidant, anticancer, antiinflammatory, antibacterial, antiviral, gastroprotective, and antiallergic properties [3,7].

Galangal is used in a variety of ways, from diet to medicine (Fig. 7.2). Galangal rhizomes are harvested all year and used in Asian cuisine because of their spicy flavor [8]. Galangal has been used in traditional medicine in many nations for centuries, in addition to its culinary uses. Because of its antispasmodic, stomachic, carminative, and antibacterial characteristics, it is used in Siddha, Ayurveda, Unani, Chinese, and Thai traditional medicine to treat stomach aches and diarrhea [2,9]. There have also been accounts of its traditional use to treat stomach pain, irregular menstruation, and hemorrhoids [2,10].

Figure 7.1
Chemical structures of important bioactive components of galangal.

The rhizome of A. galanga is widely utilized in the preparation of Siddha and Ayurvedic preparations for several diseases including heart diseases, rheumatism, renal calculus, diabetes, hypertension, asthma, ulcer, bronchitis, inflammation, microbial infections, and chronic enteritis [11,12]. The galangal rhizome, also known as rasna in India, is used in a variety of Ayurvedic remedies to treat rheumatism and inflammatory disorders, as well as other illnesses such as dementia, diabetes, and neurological diseases [13–17]. It is also identified as Kulinjan, Aratha, Rasna, or Sugandhamula. Some of the Ayurvedic formulations utilizing the rhizome of A. galanga are Rasna panchakam, Rasnadi kashayam, Rasnarendadi, and Rasnapthakam [12]. Galangal is one of the rejuvenator herbs comprised in formulations utilized to manage several chronic diseases [18]. Moreover, galangal plants have also perfumery and cosmetic applications [2].

Figure 7.2
Utilization of galangal in various functional foods and nutraceuticals with different biological activities.

Several researchers have studied galangal in depth, and a number of phytochemicals found in it, such as terpenoids, flavonoids, essential oils, saponins, and phenolic acids, have been isolated and identified [8,19]. These major bioactive constituents extracted from galangal, such as galangin, kaempferol, galangal acetate, and 1,8-cineole, have been found to validate numerous advantageous impacts on human health, specifically for inhibition of oxidative stress by eliminating free radicals, in addition to possessing antimicrobial properties, which are also usually identified as antioxidant and antimicrobial activities, respectively [5,11,20–26]. The widespread usage of galangal mainly in cuisine and traditional medicine has fascinated the consideration of scientists looking for its active compounds, therapeutic characteristics, mode of action, and possible applications [2].

7.2. Galangal in various formulations for different applications

A. galanga and A. officinarum are two of the most prevalent galangal species, and their ethnobotanical, medicinal, and nonmedicinal uses in many parts of the world have prompted numerous scientific investigations [10]. The phytochemical elements of the various galangal species have been linked to these pharmacological features [2].

7.2.1. Galangal in various functional foods

Galangal are extensively utilized as flavorings in food (Table 7.1), such as chicken and meat products, cooking food, Far East sweet wine, and bitter beer. Galangal is used as a main constituent of five-spice powder and salt-baked chicken powder [27]. Rhizomes of galangal, lemongrass, and Kaffir lime leaves are the key constituents in numerous Thai foods with desirable therapeutic effects. The rhizomes of galangal, lemongrass, and the extract of Kaffir lime leaves mainly consisted of essential oils, had advantageous effects on blood lipid profiles, histopathological changes such as oxidation, inflammation, steatohepatitis, and liver mitochondrial architecture and function. The combination has a strong chance of being developed further as a unique supplement for treating hypercholesterolemia [28]. A combination of galangal, catuaba, maca root, roseroot, guarana, and polyfloral honey can be utilized to enhance the antiinflammatory, antioxidative, and antimicrobial effects of stored sausages. Due to their advantageous effects on preventing lipid oxidation in sausages, in addition to restricting the development of microorganisms and improved antioxidant and antiinflammatory potential which may protect against degenerative disease [29].

Table 7.1

Various applications of galangal in different functional foods and packaging materials.

Application area | Usage as | Used in | Benefits | References
Functional food | Flavorings in food | Far East sweet wine, bitter beer, chicken and meat products, spice powder, salt-baked chicken powder | Improves flavor | [27]

Ingredient in functional foods

Thai foods containing rhizomes of galangal, lemongrass, and kaffir lime leaves with desirable therapeutic effects

Advantageous effects on blood lipid profiles, oxidation, inflammation, steatohepatitis, and liver mitochondrial architecture and function

[28]

Combination of galangal, catuaba, maca root, roseroot, guarana, and polyfloral honey for stored sausages | Prevents lipid oxidation in sausages, enhance the antiinflammatory, antioxidative, and antimicrobial effects | [29]
Condiment uses | Flavourings, medicinal foods and tea | Improves the appetite and stimulates the blood stream, relieves dyspepsia, vomiting flatulence, and stomach sickness | [27,30,31]
Food packing | Packaging material | Active edible film | Preservation of the vegetables and fruits | [32]
Galangal root oil/β-cyclodextrin mats | Preservation of sensory characteristics of beef | [33]

A total of 12 compounds including 8 diarylheptanoids and 4 flavonoids were identified as potential xanthine oxidase inhibitors from A. officinarum rhizome extract. Galangin, 3-methoxyl-galangin, and kaempferide were the significant xanthine oxidase inhibitors in A. officinarum rhizome extract, which can be beneficial to the potential application of A. officinarum rhizome extract in functional foods for patients suffering from hyperuricemia [34]. Rhizome of A. officinarum exhibiting stimulant and carminative bioactivities is extensively utilized in Asian and European countries as medicinal and spicey condiment usages. It is frequently utilized in flavorings, medicinal foods, and tea in China. It has the ability to improve appetite and boost blood flow throughout the body and brain. It is specifically beneficial in dyspepsia, vomiting flatulence, and stomach sickness and also utilized as a sexual stimulant [35].

Tea has a long history of being consumed as a health-promoting beverage in China. Galangal tea has positive therapeutic effect with antioxidant, antiinflammatory, hyperlipidemia, and hyperglycemia activities. Each 10g of galangal and Cyperi rhizoma tea are boiled 1h after that addition of brown sugar and adding in boiled water for oral consumption, which are supportive to chronic gastritis patients. Two-gram galangal mixed with 1g Angelicae sinensis radix, 3g Cinnamomi cortex, 1g Ginseng radix, and 2g Magnoliae officinalis cortex are boiled together, then producing tea with decoction, having the result of heating the middle stimulant to dismiss cold. A tea together with galangal and Bombyx batryticatus each 10g can exorcize wind and clear away cold, relieve pain, and disperse phlegm. It has a substantial healing outcome on cold headache [27,30,31].

The chemical composition of galangal flowers was considerably different from that of galangal rhizome. The main components of galangal flower essential oil have been identified as pentadecane and humulene. 1′-acetoxyeugenol acetate was recognized as the chief component in galangal flowers and showed the enhanced antioxidant potential and strongest antimicrobial activity because of high content of total phenols, which can be employed as a natural food preservative in the design of unique functional healthy products and could be extremely desirable and valuable [26].

7.2.2. Galangal extracts as a packaging material

An active edible film based on pullulan and carboxymethyl chitosan encapsulated with galangal essential oil was prepared to preserve the vegetables and fruits; mangoes were used as a reference fruit. The addition of galangal essential oil in pullulan/carboxymethyl chitosan composite films served as an innovative packaging material for the use of active fresh-keeping packaging for vegetables and fruits because of their substantial mechanical, biodegradability properties and protection outcome [32]. Nanofiber swim bladder gelatin nanofibers combining galangal root oil/β-cyclodextrin mats exhibited effective inhibitory outcome against EHEC O157, and significant impact on the sensory characteristics of beef, which exposed a practicable approach to explore application potential of natural antibacterial constituents in active food packaging [33]. Extracts of A. galanga and Phyllanthus emblica Linn (P. emblica). exhibited high antioxidant activities and good antimicrobial activity against the expansion of food poisoning bacteria, Staphylococcus aureus because of the major compounds of galangal extract such as β-bisabolene, 1,8-cineole, β-selinene, and β-caryophyllene and the high total phenolic content. Incorporation of these extracts in various edible films to make packaging materials can release antimicrobial and antioxidant agents to extent the shelf life of foods [9].

7.3. Biological activities of galangal for different nutraceutical applications

7.3.1. Anticancer activity

The compound, 1′-acetoxychavicol acetate was obtained from A. galanga, was shown to have higher anticancer efficacy in mice with sarcoma 180 ascites [36]. By inhibiting Rev transport, this compound also suppresses the replication of the human immunodeficiency virus type 1 (HIV-1) [37]. Other compounds, 1,7-bis(4-hydroxyphenyl)-1,4,6-heptatrien-3-one and bisdemethoxycurcumin, were isolated from A. galanga and which had an effective antiproliferative bioactivity against the human melanoma cell line and considerably prevented the proliferation of melanoma cells [38]. A compound called 1′-acetoxyeugenol acetate was isolated from A. galanga and confirmed to highly promising antitumor activity and may be beneficial as an upcoming natural product-based anticancer molecule [39,40]. Another compound, trans-3,4-dimethoxycinnamyl alcohol being one of the major phytocompound in A. galanga extract was shown to have antiproliferative effect through inducing apoptosis in human breast carcinoma cell line [41].

Breast cancer is the most common cancer in women around the world, with poor therapy. Tumor necrosis factor-related apoptosis-inducing ligand is an active antitumor agent for human breast cancer, which is combined with galangin and inhibiting human breast cancer progression in vitro and in vivo that is dependent on endoplasmic reticulum stress, reactive oxygen species, and apoptosis promotion, leading to breast cancer cells death without toxicity in normal cells or organs. The results of the study indicated that galangin and tumor necrosis factor-related apoptosis-inducing ligand combination has a potential value in suppressing human breast cancer progression [42].

Galangin combined with cisplatin at low concentrations without toxicity to cells and animals considerably induced apoptosis in A549/DDP cells via both intrinsic and extrinsic pathways by multiple targets, such as B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein (BAX), and signal transducer and activator of transcription 3 (STAT3)/nuclear factor kappa B (NF-κB) signaling pathways and efficiently attenuated tumor growth in vivo. Galangin in combination with cisplatin has the healing ability value against human lung cancer resistant to cisplatin [43]. Cancer during childhood retinoblastoma is described as a rare cancer that happens during childhood. Even though various treatments are existing for retinoblastoma, there is a necessity for alternative innovative action modalities for retinoblastoma with improved safety and efficiency profile. Galangin treatment suppressed the development of retinoblastoma tumor in vivo by apoptogenic and antiproliferative mechanisms. Accordingly, galangin might be a safe and hopeful nonchemotherapeutic drug, which could be beneficial as an adjuvant against retinoblastoma [44].

A compound, 7-(4-hydroxyphenyl)-1-phenyl-3-heptanone isolated from A. officinarum has a reasonable level of anticancer activity when related with 5-fluorouracil as control with IC_50 values less than 50μg/mL against MCF-7 and HepG2 cancer cell line [45]. Another compound isolated from A. officinarum, alpinoid-C, was found to have promising anticancer action against both human leukemia and murine melanoma (B16-F10) cancer cell lines by Reddy et al. [46]. A study was conducted to inhibit in vivo two-stage carcinogenesis utilizing mice model; it was verified that bioactive diarylheptanoids present in methanol extract of galangal revealed notable promising antitumor activity indicating the possibility of developing anticancer drugs based on bioactive components of galangal in the future [47].

7.3.2. Antiinflammatory agent

A. galanga prevented lipopolysaccharide (LPS)-stimulated inflammatory response in RAW 264.7 cells via downregulating toll-like receptor 4 (TLR4) and Janus kinase-signal transducer and activator of transcription (JAK/STAT) pathway, which determines the molecular mechanism of the antiinflammatory action of A. galanga and its healing effect for several inflammatory conditions [12]. Galangin suppresses the downregulation of inducible nitric oxide synthase (iNOS), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6) gene expression in LPS-stimulated macrophages. Moreover, the inhibition of iNOS and IL-1β expression is linked to the inhibition of the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) and NF-κB-p65 pathway in LPS-stimulated macrophages, which delivers the significant understanding about the usage of galangin as a promising healing agent in inflammation [48].

A compound, 1,7-diphenyl-4-en-3-heptanone isolated from A. officinarum has a very acceptable platelet-activating factor (PAF) receptor binding antagonist activity with an IC_50 value of 5μM that is effective to prevent PAF-mediated inflammatory diseases [49]. Honmore et al. [50] investigated that 1,7-diphenyl-5-ol-3-heptanone isolated from A. officinarum is effective to prevent the activity of cyclooxygenase-2 by binding to its active position in that way acting as an effective antiinflammatory molecule. Likewise, another compound, 3,5-dihydroxy-1,7-diphenylheptane has been described by Matsuda et al. [51], as having a very good preventive outcome on the production of nitric oxide in LPS-activated macrophages in that way downregulating the pathway leading to inflammation and multistage carcinogenesis at inflammatory locations [51]. A. officinarum is customarily utilized to manage pain, inflammation, and digestive and cold diseases. Total flavonoids from the rhizomes of A. officinarum effectively reduced the ulcer index and attenuated release of inflammatory mediators (IL-6, IL-1β, tumor necrosis factor α (TNF-α), and prostaglandin E2 (PGxE2)), enhanced the content of nitric oxide, and enhanced gastrin and motilin secretion, which can be used as a potential gastroprotective agent [52]. A Chinese patent stated that extracts of A. officinarum can be utilized for the preparation of powders, granules, tablets, capsules, pills, oral liquid formulations for clinical usage or can be injected to treat dysmenorrhea because of its antiinflammatory properties [53].

7.3.3. Cardioprotective agent

Dyslipidemia is a considerable hazard issue for cardiovascular diseases. P. emblica and A. galanga were found to have hypolipidemic effects, due to the synergistic action, which could upregulate expression of the apolipoprotein A1, low-density lipoprotein receptor, and scavenger receptor class B1 genes [54]. Cardiovascular problems are the important reason of morbidity in diabetes. Oxidative stress and inflammation are concerned in the expansion and evolution of diabetic cardiomyopathy. Galangin ameliorated hyperlipidemia, hyperglycemia, inflammation, oxidative stress, and apoptosis and prohibited cardiac damage in diabetic rats. Hence, galangin perhaps of considerable healing benefits in the treatment of diabetic cardiomyopathy and possibly other diabetes difficulties, incomplete additional studies to explore the particular fundamental mechanisms [55]. Lipid metabolism illnesses such as hypertriglyceridemia and hypercholesterolemia are vital mediators of cerebrovascular, atherosclerotic cardiovascular, and ischemic heart diseases. In this linking, it was exposed that the extracts of A. galanga are very active in attenuating the level of low-density lipoprotein in the blood serum in that way contradicting the threats linked with hyperlipidemia [56]. Obesity is currently a major public health concern and is a significant hazard influence for the origin of numerous chronic sicknesses such as coronary heart disease and hypertension. To address this problem, ethanolic extracts of A. officinarum possibly considered as having excellent antiobesity properties, subsequently can acceptably suppress body weight increase and also expand the healthy lipid profile [57].

7.3.4. Antioxidant potential

A mixture of ethanolic extract of P. emblica fruits and ethanolic extract of A. galanga rhizomes at a ratio of 7:3 showed synergistic antioxidant activities against 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonate assay, 2,2-diphenylpicrylhydrazyl assay, and superoxide anion radicals and also showed a substantial reduction in intracellular reactive oxygen species against H_2O_2-induced oxidative stress in an endothelial cell live (ECV304) in dose-dependent manner, in addition to a rise in cell survival. P. emblica and A. galanga, when combined, have the potential to evolve into novel nutritional antioxidants [58]. Galangin can modify nuclear factor erythroid 2-related factor 2 (Nrf-2) level ensuring in reduced expression of glyoxalase-1 leading to reduced detoxification of methylglyoxal. Enhanced methylglyoxal levels lead to increased levels of oxidative stress resultant in cell death. Thus, inhibitors of Nrf-2 like galangin can be related to induced cell death and decline in metastatic potential of cancer cells by reduction in glyoxalase-1 expression and enhance in injury caused by methylglyoxal and reactive oxygen species [59]. Ravichandra et al. [60] reported that galangin has substantial antioxidant action and also has the capacity to sustain cell membrane integrity, decrease oxidative stress, and remedy cardiac systolic/diastolic dysfunction in the cardiac myocytes stressed with doxorubicin [60].

7.3.5. Liver health

Galangin loaded galactosylated pluronic F68 polymeric micelles are efficiently targeted galangin to the liver attaining ∼7 folds enhance in volume of galangin in the liver when associated to galangin alone. Retaining of galangin in the liver was attained because of the controlled release, permeability glycoprotein, and cytochrome enzyme inhibition activity of pluronic F-68. Accordingly, pluronic F68-galactose conjugated copolymer can perform as a hopeful transporter structure for enhancing liver targeting of hydrophobic drugs vulnerable to permeability glycoprotein efflux and cytochrome enzyme-related metabolism [61]. Liver toxicity was induced by antitubercular medications (isoniazid and rifampicin), and ethanolic extract of A. officinarum rhizome at 200 or 400mg/kg was administrated orally once daily for 21 days. The hepatoprotective action was measured by biochemical parameters. Outcomes confirmed that the A. officinarum rhizome extracts decreased the toxicity in serum levels of hepatic enzymes. Histopathology exhibited that A. officinarum rhizome extracts decreased the hepatocellular necrosis, regeneration, and repair of cells toward normal [62].

7.3.6. Neuroprotective and memory enhancing activity

Galanga extracts were found to be powerful antiradical, antioxidant, and anticholinergic effects in various bioassays when associated to standard antioxidant and anticholinergic compounds. Galanga extracts can be utilized for lessening or inhibiting lipid oxidation in pharmaceutical products or food, postponing the development of toxic oxidation products, protecting nutritional quality, and extending the shelf life of pharmaceutical materials or nutrition. Galanga extracts had effective acetylcholinesterase (AChE) inhibition properties and can be utilized for the management of mild-to-moderate Alzheimer's disease (AD) and several other memory illnesses [63]. Hanish Singh et al. [64] explained that mice treatment with A. galanga extracts induced the acceleration of cognitive function and enhanced sodium–potassium adenosine triphosphatase (Na+/K+-ATPase), antioxidant response whereas declining AChE activity [64]. (2r, 3s)-pinobanksin-3-cinnamate is a flavanone derived from the seeds of A. galanga that showed acceptable in vitro neuroprotective properties by performing as a potential antioxidant molecule thus downregulating oxidative stress [65]. Cisplatin is a chemotherapeutic anticancer drug with serious side effects such as neurotoxicity and nephrotoxicity. Huang et al. [66] showed that galangin extracted from A. officinarum could substantially ameliorate the nephrotoxicity because of cisplatin in mice through ameliorating oxidative stress and inflammation by the inactivation of NF-κB and extracellular signal-regulated kinase (ERK) pathways [66]. Galangin defends hippocampal neurons in gerbils after ischemia-reperfusion by preventing ferroptosis by activating the solute carrier family 7-member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) axis [67].

7.3.7. Antiviral effect

The extract of A. officinarum has been utilized for management against respiratory syncytial virus, poliovirus, herpes simplex virus type 1, and measles virus in vitro. Respiratory syncytial virus infection is a dangerous sickness that affects both the children under the age of two and elderly, causing serious bronchitis and pneumonia [68]. Seven of the nine diarylheptanoids isolated from A. officinarum showed extremely strong antiviral activity against respiratory syncytial virus, poliovirus, and measles virus, according to Konno et al. [69]. In another investigation, Sawamura et al. [70] explained that diarylheptanoids, which are the bioactive constituents of galangal species showed significant antiviral activity against influenza virus under in vitro environments. Tewtrakul et al. [71] explained that methanolic extract of A. galanga showed a hopeful antiviral activity against the human cytomegalovirus and HIV-1.

7.3.8. Antimicrobial activity

Methyl cinnamate, ethyl cinnamate, and n-pentadecane were the main components of A. galanga rhizomes essential oil, which have potent antibacterial activity against enterohemorrhagic Escherichia coli O157:H7 owing to higher passive penetrability of bacterial cell membrane, subsequently crucial intracellular components efflux. In addition, A. galanga rhizomes essential oil disrupted the intracellular physiological metabolism of enterohemorrhagic Escherichia coli, together with the prevention of P-type adenosine triphosphatases activity and downregulation expression of four virulence genes related with enterohemorrhagic Escherichia coli infections [72]. Nanoemulsified essential oil of A. galanga showed higher antimicrobial activity against various microorganisms like Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Fusarium oxysporum, and
Aspergillus niger when associated with control rhizome essential oil of A. galanga. The nanoemulsified essential oil influenced considerably greater antioxidant activity as compared with the normal essential oil. Accordingly, the essential oil-based nanoemulsion of A. galanga can be utilized as an antimicrobial and antioxidant agent in pharmaceutical industries [73]. A compound, 1,7-diphenyl-3,5-heptanedione isolated from A. officinarum, which was described to have good antibacterial activity against Helicobacter pylori, indicating that it could be used as an antibacterial molecule in the future [74].

7.3.9. Reproductive agent

A. officinarum hydroalcoholic extract can expand sperm parameters such as count, motility, morphology, and viability in streptozotocin-induced diabetic rats, which is possibly mediated by antioxidant components. Furthermore, the higher dose of this extract (500mgkg
−1) appears to be further effective in the enhancement of sperm parameters and testicular damages [75]. Effect of A. officinarum rhizome extract was evaluated on spermatogram influences in men with idiopathic infertility by a prospective double-blinded randomized clinical study. A. officinarum extract exhibited a positive outcome on semen parameters in terms of sperm morphology and count in men suffering from idiopathic infertility because of its antioxidant components [76]. A compound, 1,7-diphenyl-5-hydroxy-3-heptanone isolated from A. officinarum, which has been described by Kim et al. [77], as demonstrating 5α-reductase inhibitory activity that is an important enzyme authorizable for reductive alteration of testosterone to 5α-dihydrotestosterone and its effective prevention can be a healing target for the management of androgen-dependent illnesses, particularly male pattern baldness, benign prostate hyperplasia, and acne [77]. Jakobsen and Giversen [78] filed a patent on the preparation of orally ingestible dosages of capsules/tablets containing granulate extracts from A. galanga or Alpinia conchigera containing high levels of 1′S-1′-acetoxychavicol acetate for the remedy of male infertility which is caused by low sperm count and/or by low sperm mobility.

7.3.10. Immune stimulant

Immune-stimulating action was demonstrated by hot water-soluble polysaccharide extracts of A. galanga, which greatly increased the proliferation of murine spleen cells, therefore improving the immune response efficacy of lymphocytic and phagocytic systems [79].

7.4. Conclusion and future prospective

Galangal, including greater galangal (A. galanga) and lesser galangal (A. officinarum) are widely used in various foods and traditional medicines in Southern Asia and Indonesia due to the existence of numerous bioactive components, particularly galangin, which have several pharmacological and medicinal properties such as antiinflammatory, antioxidant, anticancer, cardiovascular, liver health, neuroprotective, antiviral, antimicrobial, reproductive, and immune-stimulating activities. Galangal are extensively used as flavoring agent in foods, sausages, spicey condiments, and herbal tea. Galangal can be the promising spices to develop a variety of innovative functional foods and nutraceutical formulations that could help satisfy the necessity of the human health and wellness due to the various biological activities.

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Chapter 8: Bacopa monnieri

Nitisha Sendri
[1]
,
[2]
, and Pamita Bhandari
[1]
,
[2]

1
CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India
2
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India

Abstract

Bacopa monnieri (L.) Wettst is a well-known nootropic herb, which is commonly used in the traditional system of Ayurveda for the treatment in improving memory and reducing anxiety. Traditionally, this herb is used for several ailments, but it is mainly known as a neuronal booster and cognitive enhancer. This chapter highlights structural characterization of the active chemical constituents by spectroscopic techniques, analytical methods, and their neuropharmacolgical effects. In addition, this chapter also provides details about bioactive compounds, i.e., triterpenoid glycosides present in B. monnieri, and shows chief therapeutic action in neuroprotection. The traditional claims of B. monnieri are authenticated by reporting numerous preclinical and clinical studies ensuring the specialized hydroalcoholic extract of B. monnieri and standardized phytochemical profile. Numerous evidence suggests that B. monnieri acts via resulting mechanisms—antioxidant neuroprotection (inhibition of reactive oxygen species and enzyme viz., poly [AAP-ribose], polymerase [PARP], and prolyl endopeptidase [PEP]), β-amyloid reduction, increased cerebral blood flow, acetylcholinesterase inhibition and/or choline acetyltransferase activation, and neurotransmitter modulation (acetylcholine, dopamine, 5-HT, serotonin). Thus, reviewing prolonged effects of a standardized extract of B. monnieri on neurological function in human subject for the purpose of standardized extract has been popularly commercialized in the names of bacognize, bacomind, synapsa, keenmind, and mediherb.

Keywords

Bacopa monnieri; Commercialized formulations; Functional food; Neuroprotection; Nootropic herb; Triterpenoid saponins

8.1. Background

Bacopa monnieri (L.) Wettst (Brahmi, water hyssop, Indian pennywort), belonging to the family Plantaginaceae, is a perennial, creeping herb found throughout the wet lands of Indian subcontinent, particularly northeast and southern regions. The Bacopa genus comprises around 100 species of aquatic herbs with a distribution throughout the warmer regions of the world [1]. The logic behind its common name was that our brain is the creative center, and any product that finds improvement in the brain condition is called Brahmi [2]. In Indian Ayurvedic prescriptions, B. monnieri has been in use for approximately 3000 years back and categorized as “medhya-rasayana” where “medhya” (in Sanskrit) means intellect or cognition and “rasayana” stands for rejuvenation. Medhya rasayana is a medication that eases the impacts of mental stress and further enhances brainpower and cognitive function [3]. Traditionally, it has been used as a nerve tonic to improve the memory revitalizing capacity, attentiveness, as well as to reduce anxiety [4]. It has also been documented in ancient articles of Ayurveda including Charaka Samhita (Sixth century AD) and Bhavprakash Var-Prakarana (16th century AD) [5]. Numerous Ayurvedic formulations recommended for cognitive decline consists of B. monnieri as a primary component. B. monnieri possesses high-value medicinal properties such as neuroprotection against dementia, amnesia, Parkinson’s disease (PD), schizophrenia, Alzheimer’s disease (AD), and epileptic seizures. It has also been documented to exhibit other pharmacological activities such as antiinflammatory, antimicrobial, anticonvulsant, antiageing, anticancer, antidepressant, antiemetic, broncho-vasodilatory, and antiulcer activities [6]. This ancient medicinal plant contains spectrum of bioactive phytochemicals such as triterpenoid glycosides [7], cucurbitacins [8], sterols [9], phenylethanoids [10], and alkaloids. Numerous studies have documented on cognition-enhancing effects of whole plants or alcoholic extract of B. monnieri, including improved retention of memory, motor learning, and acquisition. The extracts of B. monnieri have been widely examined for their neuropharmacological effects. The cost of this plant continued to increase day by day due to its potential uses against numerous neurological disorders including AD. The National Medicinal Plants Board of India had estimated the annual market demand at 1000 tons in 2000 year [11]. The traditional claim of B. monnieri being nootropic herb has been scientifically proved. Besides its therapeutic significance, it is also a famous ornamental plant [12]. Conventional methods have a slow rate of propagation; thus, few tissue culture procedures have been accounted for in vitro production of this curative plant.

8.2. Plant morphology

B. monnieri is a small, semisucculent herb that thrives on wet soil or in very shallow water. Its leaves are typically 5–25mm long, narrowly spatulate, or oblong in shape with white to pink or purple flowers. It is mainly found in Indian subcontinent, Southeast Asia, Australia, tropical Africa, and Subtropical United States [13]. It is a creeping herb of height 2–3 feet with branched and small fleshy, oblong leaves. This creeper plant occurs simultaneously with fruits in summer [2] and found at 1500m altitudes, and clayey loam to clayey soils are suitable for the propagation. In North Indian region, it can propagate in the range of soil pH (5–7.5) and temperatures (15–40°C).

8.3. Chemical constituents

A systematic chemical examination uncovered that B. monnieri contains bacoside A (64.28%), bacoside B (27.11%), D-mannitol (0.83%), stigmastanol (2.08%), betullic acid (4.58%), β-sitosterol (0.58%), and stigmasterol (0.54%) (Basu et al. 1967; [14]. However, at that time, the composition of bacosides A and B was not determined.

Dammarane triterpenoid glycosides (bacosides, bacopasides, or bacosaponins) are the major isolated chemical constituents of ethanolic extract of B. monnieri and characterized to be responsible for memory enhancing and other neuroprotective effects. Bacosides are extensively studied for their potential effects in neuroprotective activity. Bacosides are the characteristic chemical constituents contained aglycone, i.e., jujubogenin (1) or pseudojujubogenin (2) and sugar unit (Fig. 8.1).

The linkage of isobutylene unit to aglycone (1) or (2) distinguished the type of skeleton. In aglycone 1, the isobutylene unit is attached to C-23 position of skeleton, whereas the isobutylene unit is linked to C-22 position in aglycone 2. The glycan part includes α-L-arabinopyranosyl, β-D-glucopyranosyl, α-L-arabinofuranosyl, and 6-O-sulphonyl-β-D-glucopyranosyl. Bacoside A and bacoside B are extensively studied for their potential effects in neuroprotective activity. Previously, it was considered that bacoside A is a single chemical entity named as 3-(α-L-arabinopyranosyl)-O-β-D-glucopyranoside-10,20-dihydroxy-16-keto-dammar-24-ene [14] and documented to coexist along with bacoside B which varies in optical rotation [15,16]. Later on, a high-performance thin-layer chromatography (HPTLC) method [17] and high-performance liquid chromatography (HPLC) technique have been developed for the quantitative determination of bacoside A and found to be a combination of four triterpenoid glycosides (3–6) [18]. Bacoside A was considered as the main active constituent for cognitive-enhancing activities, and its chemical composition was established as a combination of four triglycosidic saponins, namely, as bacoside A_3 (3), bacopaside II (4), bacopaside X (5), and bacopasaponin C (6) [18,19]. Similarly, bacoside B composed of four diglycosidic saponins, i.e., bacopaside N_1 (7), bacopaside N_2 (8), bacopaside IV (9), and bacopaside V (10). Apart from bacosides, other chemical constituents have also been classified in B. monnieri, i.e., hersaponin, apigenin (11), luteolin (12), plantainoside B, D-mannitol, cucurbitacin, monnierasides I–III, and alkaloids as brahmine (13), nicotine (14), and herpestine (Fig. 8.2) [20]. The triterpenoid glycosides, i.e., bacopasides I–XII [21] bacoside A_1-A_3 [22–24], bacopasaponin A-G [25–27 ], have been identified in herbal extract of B. monnieri. Bacosine (15) [7] has been identified as a free triterpene from B. monnieri [3].

Figure 8.1
Structures of jujubogenin (1) and pseudojujubogenin (2) aglycones.

8.4. Extraction methods for chemical constituents

The classical extraction procedure for obtaining active constituents from the plant material is the use of soxhlet and maceration with a ratio of methanol/water or ethanol/water and partitioning with n-hexane followed by chloroform/ethyl acetate and n-butanol to have extracts of differed polarities in sequence [8–10,28 ]. Mostly, the plant material is defatted with n-hexane preceding to extraction [26,29].

Preliminary phytochemical analysis by qualitative chemical tests is a major concern for the detection of active principles, present within it. Right after the extraction procedure, researchers were used to subject different fractions of B. monnieri extracts to preliminary qualitative tests to get the idea about the active phytochemicals. This preliminary screening gives the ideas about the presence of active constituents such as saponins, alkaloids, flavonoids, and steroids [30]. A positive preliminary Liebermann–Burchard (L.B.) test and Molisch test for triterpenoids and sugar detection were used to analyze prior to column chromatography, respectively [31]. The n-butanol fraction was purified by using the combinations of silica gel adsorption column chromatography and preparative thin-layer chromatography (TLC) followed by crystallization to yield major active dammarane triterpenoid glycosides [6]. Some of the saponins were isolated from macroporous resin chromatography followed by Sephadex LH-20 and reverse phase (RP-C_18) HPLC columns and thereafter purified by semipreparative HPLC column chromatography [32]. Besides n-butanol fractions, the ethyl acetate fraction on repeated silica gel chromatography [22] or over Diaion HP-20 [33] yielded bacopaside-XI and bacopaside-XII.

8.5. Analytical techniques used for qualitative and quantitative analysis of B. monnieri

Besides the isolation, identification, and characterization, quantification of triterpenoid glycosides was conducted by using different analytical techniques. Various techniques used for the investigation of specialized metabolites in B. monnieri extract includes TLC, HPTLC, supercritical fluid chromatography (SFC), HPLC, nuclear magnetic resonance (NMR), and liquid chromatography mass spectrometry (LC-MS/MS). An RP-HPTLC and SFC analytical methods were developed for the standardization and routine analysis of compounds 3 and 16 in B. monnieri extract [34] and its monoherbal capsule formulations [35]. Afterward, a rapid, simple, and economical HPTLC method was developed and validated for the quantitative determination of bacoside A in leaves of B. monnieri, which was collected from different geographical regions of India [36]. The conflict in the structures of bacosides A and B were attempted to delineated their complex chemical structure and found to be mixture of four saponins [18,37]. The estimation of total saponin content in B. monnieri extract and formulations were investigated by simple and sensitive HPLC method and revealed 12 bacopasaponins viz., compound 3, 4, 5, 6, 8, bacopaside I (16) and the minor components were 7, 9, 10, bacopaside III, bacopasaponin F, and bacopasaponin E [38]. A developed HPLC method determined that approximately one-third part of the concentration of total saponin in B. monnieri was identified as bacopaside I [19] and confirmed to consists mixture of four individual saponins (3–6) in bacoside A. HPLC-ELSD method was developed on five different C-18 HPLC columns and observed that best resolution and rapid separation was achieved on silica-based Chromolith column. The method was reported by Bhandari et al. [39] and has been developed for the determination of bacosides (bacoside A3, bacopaside I, bacopaside X, bacopasaponin C, bacopaside II) and apigenin. Similarly, the authors have also standardized a microanalytical HPLC method for the determination of bacoside A in small quantities of plant material, i.e., ≥2mg [40].

Figure 8.2
Structures of active constituents influence neuroprotection.

8.6. Quantitative analysis of bacosides

The method for the quantification of bacosides has been developed via ultraviolet spectroscopy [41] and HPLC [18,19]. Various bacopa products are marketed as drug and food supplementation with claims of containing specific content of bacosides but remains a lack of data in regards to the quantification and stability of these active saponins in the whole plant material. The collection of bacoside A in samples of B. monnieri from different locations was well validated and quantified [18]. Similarly, Srivastava et al. [42] revealed the stability parameters of crude extract and its posteffect on quantitative analysis of active individual compounds viz., bacoside A and bacopaside I. The techniques like LC coupled with MS and NMR-MS had additionally been utilized for rapid screening of active components. Furthermore, for the determination of active saponins from B. monnieri Wettst, the combined structural information from LC–NMR and LC-MS using LiChrosphere C-18 analytical column and mobile phase was acetonitrile-water at a flow rate of 0.8 mL/min [43]. The qualitative analysis clearly shows an insight into the existence of bacoside A_3, 3-O-β-[β -D-glucopyranosyl (1→3)-O-[α-
L-arabinofuranosyl-(1→2)]-O-β-D-arabinopyranosyl) oxy] jujubogenin, 3-O-[β-D-glucopyranosyl(1→3){α-L-arabinofuranosyl-(1→2)}-α-L-arabinopyranosyl] pseudojujubogenin, and bacopasaponin C. Different MS techniques including electrospray ionization (ESI)-ion trap (IT), atmospheric pressure matrix-assisted laser desorption/ionization (APMALDI-IT), high-energy collision-induced dissociation (CID), postsource decay, MALDI-IT/reflectron time-of-flight-MS, and low-energy CID analysis were used to understand the glycosidic linkages and distinction of aglycone for the analysis of isolated saponins from B. monnieri [44]. The molecular mass of the dammarane triterpenoid glycosides was mainly determined by using the two different MS techniques in positive ion mode (high-resolution-ESI-time of flight-MS and fast atom bombardment).

However, the applied MS techniques were incapable to distinguish the type of aglycone (1 and 2). Furthermore, an LC coupled with ESI-QTOF-MS was conducted in structural characterization of dammarane triterpenoid glycosides in B. monnieri. This systematic study reported to distinguish aglycone, i.e., isomeric jujubogenin and pseudojujubogenin [45] where their glycosides displayed the parent ion [M]+ peaks at m/z 455 [M+H–H_2O]+ and m/z 473 [M + H]+, respectively.

8.7. Structure elucidation of saponins in B. monnieri

Structure elucidation of active saponins includes the investigation of both the moieties, i.e., aglycone and sugars, which becomes complex with the increment of sugar units and its branching points. Therefore, the utilization of chemical procedures along with standard spectroscopic techniques for entire clarified structures is required. The dark blue spot of tritepenoid glycosides could be visualized on TLC plate by spraying either anisaldehyde, L.B. reagent, or vanillic-sulfuric acid reagent and heating provided a clue to the terpene character of the compound. Being a complex structure, dammarane triterpenoid glycosides can be predicted through infrared (IR) spectroscopy, MS data, and for the detailed structure elucidation, NMR spectral techniques are essential.

8.7.1. Infrared spectroscopy

IR spectrum of active saponins exhibited an absorption band close to 1635cm
−1 corresponding to have olefinic bond, and the characteristic band obtained at 1220 and 809cm
−1 shows the existence of sulfate group. It also exhibits strong and broad absorption band at 3320–3600cm
−1 analogous to multiple hydroxyl groups in the compounds.

8.7.2. NMR spectroscopy

Dammarane triterpenoid glycosides, the active constituents of B. monnieri belong to tetracyclic ring and exhibited seven methyl singlets in 1H NMR spectrum. Their structural characteristics are with tertiary methyls Me-18, Me-19, and Me-21, which are always in β-orientation and α-orientation, respectively, thus directing S-configuration to C-20 position. The differentiation between jujubogenin and pseudojujubogenin aglycones can be seen by the chemical shift of methylene proton H-22, methine proton H-23, C-17, C-20, C-21, C-24, and C-25. The IH NMR-spectrum of H-22 resonates at ≈δ
H
1.63–δ
H
1.73 whereas H-23 resonates at nearby δ
H
5.18, in case of the jujubogenin aglycone. The 13C NMR spectrum shows chemical shifts for C-17, C-20, C-21, C-23, C-24, and C-25 at δ
C
54.0, δ
C
68.5, δ
C
30.0, δ
C
68.0, δ
C
127.0, and δ
C
134.0, respectively [27,46]. In IH-NMR of pseudojujubogenin aglycone, the downfield signal of H-22 was at≈ δ
H
2.57, while upfield signals of H-23 (two protons) were at ≈δ
H
3.86 and 4.70. Further, in 13C NMR, significant upfield signals of C-23, C-24, and C-25 were observed which confirmed the aglycone as pseudojujubogenin [26,27,33]. 1H-NMR spectrum displayed olefinic signals at≈ δ
H
5.84 for H-24 proton as a broad doublet. In 13C-NMR spectroscopy, two olefinic signals were observed for C-24 and C-25 of isobutylene group with the characteristic downfield signals at δ
C
127.0 and δ
C
134.0 (aglycone 1) and ≈2–3ppm upfield (aglycone 2), respectively. The 13C-NMR spectrum displayed signals at δ
C
110.3 and δ
C
71.8 corresponding to ketal group at C-16 and methylene carbon (C-30) in aglycone unit, respectively. The structure of jujubogenin aglycone was further confirmed with the key cross-peak correlations in heteronuclear multiple bond correlation (HMBC). The HMBC correlations between the methylene signals of H-22 with carbons (C-17, C-20, C-21, and C-23) and protons of H-23 with C-24 and C-25. The glycosidic linkage to the particular position (C-3 and C-20) of aglycone was confirmed by downfield shifting in ≈ 8–9ppm at C-3 and approximately 4–5ppm at C-20 in 13C-NMR [31,37]. The configurations in the aglycone 2 were determined to be 20 (S) and 22 (R) for C-20 and C-22 by ROESY experiment.

The complete determination of sugar units present in dammarane triterpenoid glycosides of B. monnieri includes the identification, interglycosidic and sugar unit linkage to the aglycone. Preliminary the sugar constituents present in the saponins were analyzed by TLC of their acid hydrolysates with reference to sugar standards and further with NMR spectroscopy. In the dammarane triterpenoid glycosides, the chemical shifts obtained for glycosidic proton are δ
H
≈3.93–6.18 and for carbon are δ
C
98.7–110.1. Glycosylation at hydroxyl group of C-3 position results in downfield shifting of H-3 (≈δ
H
3.16–3.31) and C-3 (δ
C
75.1). The attachment of sugar units to the aglycone, interglycosidic linkages, and 1H & 13C NMR signals of the specific sugar moieties were further confirmed from 2D NMR experiments viz., 1H–1H COSY, HMQC, ROSEY, and HMBC correlations [27,29,47,48 ].

The anomeric proton signals of carbohydrate unit observed around δ
H
4.51–5.05 with coupling constant (J) value 8.0Hz suggested β-D- glucopyranosyl moiety and the anomeric proton of α -L-arabinopyranosyl had ≈δ
H
4.76–4.85 as a broad singlet with J value ≈5.8–7.0Hz. 1H NMR signals of α-L- arabinofuranosyl moiety was observed at δ
H
≈5.41–6.27 as doublet with J value< 3 [25,27,37,46 ]. Besides these sugar units, compound 16 had sulfate group which is linked to C-6 of the glucopyranose unit. A careful study at the 1H and 13C chemical shifts of compound 16 revealed that the methylene proton (H_2-6) and carbon (C-6) experienced deshielding by>0.5 and 4.5ppm, respectively and further justified by the observed upfield shift of C-5 by≈2.3ppm of glucopyranose because of γ-effect [29]. In an occurrence of trisaccharide in triterpenoid glycosides, two kinds of interglycosidic linkages, i.e., 1→2, one terminal sugar moiety is attached to C-2′ of inner sugar unit and 1→3, another terminal sugar unit is appended to C-3′. Bacopaside II is a trisaccharide of aglycone 2 consisting of arabinofuranose and glucopyranose as terminal sugars attached to C-3′ and C-2′, respectively, with inner glucopyranose by means of HMBC correlations. The cross peaks correlated between anomeric proton H-1ʺ (δ
H 6.24) of arabinofuranose with C-2′ (δ
C
79.1) of the inner glucopyranose and H-1ʺ (δ
H 5.18) of terminal glucopyranose with C-3′ (δ
C
88.7) of the inner sugar unit. Also, the HMBC experiment between anomeric proton (H-1′) of inner glucopyranose correlated to C-3 of pseudojujubogenin aglycone showed that the trisaccharide moiety is linked to C-3 of aglycone via inner glucopyranose unit [29].

The carbons exhibited a significant downfield shift included in the glycosidic linkages and are used to know interlinkages between the sugar units. Bacopasaponin C (a trisaccharide pseudojujubogenin containing terminal sugars [glucose and arabinofuranose] and inner sugar [arabinopyranose] unit) displayed C-3 linkage to aglycone 2 assigned by HMBC correlations between arabinopyranose H-1ʹ (δ
H 4.76) and C-3 (δ
C
88.6) of aglycone 2. A significant downfield chemical shift was observed for C-2′ and C-3′ of arabinopyranose from δ
C
(76.7ppm) by∼2ppm and (δ
C
83.1ppm) by∼5–6ppm, respectively. The characterization of terminal sugar units can be specified by the absence of any glycosylation chemical shift.

It has been documented that bacosides are responsible for the improved mental performance and retention capacity. In order to authenticate the traditional claims for B. monnieri, numerous preclinical and clinical studies were investigated and patented the whole extraction process that also includes manufacturing process to ensure the standardized phytochemical profile and its therapeutic action. The stability of a jujubogenin glycoside, bacoside A_3 and pseudojujubogenin glycoside, bacopaside I in ethanolic dried extract was also investigated. For the manufacturing of extract, it can be processed at a temperature less than 60°C and stored at low temperature under dry conditions to maintain the stability of markers compounds [49]. The preformulation analysis of the extracts could be beneficial for product formulation (tablets, capsules), estimate shelf life and transportation conditions for the quality assurance of the final products.

8.8. Biological activities

B. monnieri has been shown to modulate hypothalamic–pituitary–adrenal (HPA) axis activity [50], activates expression levels of brain-derived neurotrophic factor (BDNF) signaling pathways [51], changes in neurotransmitter (dopamine and serotonin) concentrations [52], antioxidant and inflammatory activity [53,54]. Specifically relating the pathophysiological processes with sleep problems, B. monnieri has association with improved sleep patterns and mood enhancer due to relation between oxidative stress, inflammation, HPA axis activity, and concentrations of neurotransmitters. B. monnieri has also been described to possess sedative, antimicrobial, calcium antagonistic, antinociceptive, anticonvulsant, antiaging, cognitive enhancive, anticancer, hepatoprotective, broncho-vasodilatory, antiallergic, antiemetic, antihypercholesterolemic effect, antidepressant, and antiulcer activities [55–57 ].

Bacosides/bacopasides were accounted for the reversal of morphine hyperactivity [58], hepatocarcinogenesis [59], memory loss in epilepsy [20], oxidative stress [60], inflammation [61], and hepatotoxicity [59]. The extract of B. monnieri is found to have potential beneficial effects against depression [62] to restore the regular level of BDNF, cyclic-AMP response element binding (CREB), Akt (protein kinase B) on chronic unpredictable stress (CUS)-induced behavioral depressive animal model [63].

Traditionally, the extracts of B. monnieri have been utilized to treat various ailments for centuries (epilepsy, pain, and inflammation), but the chief therapeutic action is in improving memory. The traditional use as a memory-enhancing effect [64] was verified with its neuroprotection, nootropic action, and its effective prevention in cognitive decline [65]. Besides cognitive enhancer, it also helps in the improvement of concentration, recall, comprehension, sharpness and is additionally known as a natural antioxidant concerned to neuroprotection [66]. Of the most recognizable therapeutic effects of B. monnieri, researchers mainly interested at the neuroprotective action against memory dysfunction, amnesia, AD, dementia, epileptic seizures, PD, and schizophrenia. From centuries, bacopa is recognized for its neuroprotective effects; therefore, we will discuss the overall neuroprotection part. B. monnieri is perhaps one of the most scientifically studied in terms of mechanisms of action.

8.9. Neuroprotection

Numerous in vivo studies and clinical trials have been performed by various researchers for remarkable cognitive-facilitating, antidementic, and acetylcholinesterase (AChE)-inhibiting effects of B. monnieri [67,68]. The studies had been carried out with alcoholic extract of B. monnieri and showed that memory-enhancing effect was because of the presence of bacoside A and bacoside B [69,70]. Besides memory-enhancing activities, bacosides also helps in the reparation of damaged neurons by upregulating kinase activity and also suppress the amnesic effects against scopolamine-induced memory impairments [71]. The mechanistic restoration action has been considered that bacosides instigated dephosphorylation membrane with development in protein and RNA turn over in specific brain zones [72]. Conclusively, therapeutic significance of B. monnieri and its active component bacoside-A is shown by the reversal of GABA receptor alteration binding in cerebral cortex neurons, associated mood disorders, and memory troubles during epilepsy [66]. Bacoside A showed neuroprotective action on β-amyloid-induced cell death in SH-SY5Y cells, which was possibly due to its capability to suppress cellular AChE activity and antioxidant posteffects nature [73]. Bacoside A significantly showed inhibitory effects upon with β-amyloid membrane-induced cell toxicity, the peptide which plays a prominent role in AD [74]. A supplementation of bacoside A and bacoside B upregulated dopaminergic imbalance results in reduced activity in the cerebral cortex of hypoglycemic rats. The neuronal damage in the cortex via downregulating SOD expression in the early stages of brain development can drastically affect memory and cognition. The clinical significance of B. monnieri extract and bacoside A pointed to the importance of a proper resuscitation program to overcome hypoglycemia [75].

Besides bacosides, compound 16 has beneficial effect on memory enhancing and neuropathology demonstrated in APP/PS1 transgenic mice. The possible mechanism of AD was explored for the first time using a treatment of compound 16 significantly promotes β-amyloid clearance via the initiation of suitable degree of inborn immune-facilitated stimulation and phagocytosis [68]. A major compound 16 in B. monnieri extract influences neuroprotection in ischemia-induced memory deficits via restoring phospho-Akt level (an antiapoptotic factor) [76] and significant effect of neuroprotective activity against middle cerebral artery occlusion–induced cerebral ischemic injury in rats provided by histopathological findings [77]. In in silico study, bacopasaponin G (17) and bacopasaponin N2 (18) showed inhibitory effects on tau protein kinase I and caspase-3 receptors to prevent AD therapy [78].

Bacoside A also showed therapeutic potential in attenuating the changes against PD induced by 6-hydroxy dopamine [79], while its component bacoside A_3 and bacopaside II showed comparatively higher reduction in changes against H_2O_2-induced oxidative stressed neuronal (N2a) cells [80]. The infusion tea of bacopa and bacoside A offers significant reduction in neuroinflammation and offers potential source for the treatment of central nervous system (CNS) disorders including depression, AD, and schizophrenia by inhibiting caspase 1 and 3 receptors and matrix metalloproteinase-3 in the cell-free assay [53]. Overly, bacoside A has been found to be therapeutic for the treatment of neurodegeneration, where oxidative stress and β-amyloid are considered as the major factor in the initiation and elevation of neurodegeneration in AD. Whereas, targeting such therapeutics to the CNS is restricted by mechanisms introduced at the blood–brain barrier (BBB). In such case, bacoside A nanoparticles (NPs) targeted the brain and had crossed the BBB [81]. Similarly, nanosized B. monnieri extract coated platinum NPs reversed the MPTP-instigated neurotoxicity in experimental parkinsonism zebrafish model via suppression of mitochondrial complex I and antioxidant action [82]. Bacosides were found to be well tolerated by healthy human volunteers and suggested to increase higher order cognitive actions. In clinical trials of 40 school children aged 6–8 years, B. monnieri (1g powder or in syrup form daily) improved memory and reaction performances without having any side effects for three months [83].

8.10. Examples of standardized extract of B. monnieri as herbal formulations

CDRI-08 is a nootropic well-characterized alcoholic extract of herb B monnieri with major constituents, i.e., bacoside A 64.28% and bacoside B 27.11%. The bacoside A-enriched standardized extract (containing a minimum of 55%±5% of bacosides) of B. monnieri was developed and evaluated to have significant facilitatory effect on acquisition, consolidation, and retention [84]. CDRI-08 was also found to be effective in attention deficit hyperactive disorder (ADHD) treatment of children [85], concentration [86], and age-associated memory impairment observed in elderly patients [87]. This product was efficacious on hippocampal-dependent contextual memory [88]. Since the findings were observed in experimental animal models, specific CDRI-08 extract of B. monnieri has been also attained a meaningful pharmacological and physiological effect after human clinical trials. This product has been researched in multiple randomized controlled trials validating that it can help in the improvement of cognitive functions and is well accepted at the recommended dose [87,89–93 ]. The first molecular evidence of the recovery potential of CDRI-08 is in the amnesiac effect of scopolamine via downregulation of neuronal and glial plasticity markers. The evaluation of pre- and postadministration of CDRI-08 extract also reduced AChE activity [94]. Rai et al. [95] investigated the effects of CDRI-08 in restoring the spatial memory of the scopolamine-treated amnesic mice and provide the evidence for the underlying mechanism. A daily dose of CDRI-08 (200mg/kg body weight) restored spatial memory by reducing AChE activity and upregulating ionotropic glutamate receptors (GluN2B) subunit expression in hippocampus as well as prefrontal cortex to amnesiac mice for seven days.

The exploration for an effective herbal drug to increase memory acquisition, retention, and learning is escalating. Dozens of compounds have been investigated by pharmaceutical industries to prepare “cognitive enhancers” that can avert cognitive decline and retained learning ability. A few herbal dietary supplements that contain active components marketed for their capability to support brain and nerve function and improve memory. Among traditional Indian medicinal herbs, B. monnieri is one of the most notable modern herbal nutraceuticals aimed to boost mental health, memory, and alertness. The systematic standardized process for the preparation of bacosides-enriched fraction from the herb B. monnieri overcomes the known neurological disorder.

In clinical trials, B. monnieri has been determined to have positive cognitive effects [83,92,96] and also suggested that it may offer on emotional well-being opportunities [97]. Similarly, Bacognize is another standardized extract of the whole herb B. monnieri, which was standardized by the United States Pharmacopeia using HPLC to total 12% of B. monnieri glycosides, i.e., bacoside A (compounds 3, 4, 5, 6) and compound 11 [97]. Recently published 150mg capsule of Bacognize supplementation (twice daily) increased 18% in morning alertness and positive results on mood and stress-related hormones [97]. The HPLC-standardized B. monnieri extract (Bacognize) has also been shown to be safe on elements of cognitive functions when administered 300mg capsules to 20 healthy older adults for 12 weeks [83]. In another study, Bacognize was evaluated by the effect on the memory of medical students. A daily administration (300mg) of Bacognize for six weeks presented a significant improvement in cognitive functions [96]. In contrast to other marketed products based on CDRI-08, Bacognize is standardized for SABG's (Serotonin Active Bacopa Glycosides). Hence, Bacognize clinically proven for adaptogenic support targeting memory, learning, focus, attention, cognitive wellness, mood, emotional balance, stress management, and circadian rhythm support for improved quality of life and quality of sleep. Table 8.1 covers a selection of key clinical studies on commercialized bacoside-rich B. monnieri extract.

BacoMind is a standardized phytochemical composition with reference to bioactive compounds derived from B. monnieri. BacoMind was developed from enriched phytochemical composition of B. monnieri, which is different from the other well-characterized and commercialized standardized extracts. Standardization of BacoMind to nine different active constituents up to 50% by weight of the total composition (w/w) viz., compound 3 (>5.0%), compound 16 (>7.0%), compound 4 (>5.5%), jujubogenin isomer of bacopasaponin C (>7.0%), compound 6 (>4.5%), bacosine (>1.5%), compound 12 (>0.2%), compound 11 (>0.1%), and β-sitosterol-D-glucoside (>0.3%) [104]. The individual active component present in BacoMind was quantified and found to be contained bacoside A_3 (80mg), bacopaside II (68mg), bacopasaponin C (52mg), jujubogenin isomer of bacopasaponin C (90mg), bacopaside I (89mg), bacosine (23mg), luteolin (4mg), β-sitosterol-D-glucoside (9mg), and apigenin (2mg) per gram on dried weight basis [105].

Table 8.1

Clinical studies on commercialized bacoside-rich Bacopa monnieri extract.

Clinical trials studies on CDRI-08
Intervention | Participants/study design | Experimental time | Clinical outcome | References

300mg/day

46 healthy adults (18–60 years age group)

12 weeks

Significant improvement in speed of visual information processing learning rate of memory consolidation and significant reduction in the rate of forgetting

[93]

300mg/day | 76 healthy adults (40–65 years age group) | 3 months | Retention of new information | [91]
300mg/day | 107 healthy adults (18–60 years age group) | 90 days | Significant improvement in working memory, attention, and accuracy in complex cognitive tasks | [92]
300mg/day | 40 adults with memory concerns | 12 weeks | Significant improvement in mental control logical memory paired associated learning | [87]
300mg/day | 17 healthy adults | Two hours | Positive effect in acute cognitive function mood stress | [89]
Two doses 320 mg/640mg | 24 healthy adults | 2 hours | Improved cognitive performance including decision-making time and attention or freedom from distractibility | [90]
BacoMind
300mg for first 15 days and 450mg for next 15 days | 23 healthy volunteers (8–45-year age group) | 30 days | Found to be safe and tolerable; minor gastrointestinal side effects were reported | [98]
300mg daily | 98 healthy participants (upto 55 years age group) | 12 weeks | Improved memory acquisition, retention, and learning with side effects (stool frequency, nausea, and abdominal cramping) | [99]
Table Continued

Clinical trials studies on CDRI-08
Intervention | Participants/study design | Experimental time | Clinical outcome | References
450mg | 65 age-associated memory impaired participant (65-year age group) | 12 weeks | Improved cognitive functions such as verbal memory and attention | [100]

225mg

27 children with ADHD (6–12 years)

6 months

Reduced symptoms of ADHD in all children, i.e., restlessness, self-control, learning problems, impulsivity, and psychiatric problems

[101]

225mg | 24 children (4–18 years) | 4 months | Improved in the battery of memory test (short-term verbal memory, working memory, visual memory, mental control) | [102]

Mediherb

300mg

48 healthy participants (74-year age group)

12 weeks

Potential for safely enhancing cognitive performance in the aging

[103]

Bacognize
300 mg/day | 50 participants (18–70 years) | 28 days | Improvements in physical and emotional health; increase in morning alertness | [97]

300mg capsule/day

20 healthy older adults (60- and 75-year age group)

12 weeks

Safe on elements of cognitive functions;significant improvement in short-term memory, processing speed, attention, and depression

[83]

300 mg/day | 60 medical students | 6 weeks | Significant improvement in the tests relating to the cognitive functions | [96]

Henceforth for the establishment of BacoMind safety, four human efficacy clinical trials were undertaken. In the context to extensive safe use of this herb, BacoMind was evaluated for toxicological studies to confirm its safe usage. An oral administration of BacoMind in Sprague–Dawley rats did not show any sign of its toxic nature with respect to body weight gain, clinical signs, neurological examination, hematological and blood biochemistry parameters up to a dose of 500mg/kg body weight. The other parameters, i.e., histopathological examination and necropsy also did not show any significant changes [106]. Further, BacoMind was assessed for clinical safety and tolerability at given doses [98], and the results showed that BacoMind did not cause any remarkable physiological, electrocardiographic, or biochemical untoward effects in volunteers at the given oral dosage of 300 and 450mg. In a preclinical trial, BacoMind significantly enhanced acquisition and retention of memory in normal and cognition-impaired scopolamine-treated mice [107]. Similarly, Morgan et al., studied the effectiveness of BacoMind and significantly improved memory acquisition and retention in healthy elderly individuals with minor gastrointestinal tract side effects. The reason of side effects theorized to be caused by the cholinergic effect of the herb [99]. In randomized double-blind placebo-controlled clinical trial, BacoMind showed significant improvement in cognitive functions such as visual retention, verbal memory, and attention in age-associated memory-impaired elderly individuals [100]. It is also found to be effective for children (6–12 years) with ADHD symptoms. A dose of 225mg/day for six months reduced symptoms of ADHD and responded with reduction in restlessness, impulsivity, psychiatric problems and improved learning problems, self-control [101]. Hence, clinical studies suggest that BacoMind indeed has the efficacy to improve ADHD symptoms in children, cognitive enhancer for children [102], and memory performance in elderly participants [100]. The molecular mechanism(s) of cognition-enhancing activity of this well-characterized marketed product possibly via finding routes viz., dopaminergic pathway by inhibiting catechol-O-methyl transferase, serotonergic pathway by antagonizes 5-HT_6 receptors, and neuroprotective mechanisms, including inhibition of reactive oxygen species (ROS) and enzyme viz., poly (AAP-ribose) polymerase and prolyl endopeptidase [108].

Calabrese et al. [103] studied the effect of dry extract of Bacopa standardized with minimum 50% bacosides (Mediherb) on 48 elderly healthy participants. A daily dose of 300mg for 12 weeks had improved various cognitive paradigms viz., Stroop task reaction, delayed recall memory as well as experienced reduction in depression and anxiety. Overly, these conducted clinical studies suggest that bacoside-rich B. monnieri extract indeed has the ability in improvement of cognitive functions as well as helps individuals respond in mental conditions including anxiety and stress.

The mentioned products were developed after careful evaluation and efficacy through animal studies and clinical trials. As a cognitive supplement, B. monnieri extracts based on a several patented formulas that follow a standardized level of quality. There are various natural memory improvement formulations available in the market based on bacosides-enriched standardized extract of herb B. monnieri. One of the formulations is CDRI-08, which has been developed by the CSIR-Central Drug Research Institute (CDRI) in India and successfully commercialized worldwide. CDRI is responsible for formulating such a standardized Bacopa extract that adequately reflected each individual component in B. monnieri. CDRI-08 is a specialized hydroalcoholic extract of B. monnieri that incorporates stems, roots, and leaves harvested at a particular point on time in order to optimize its bacoside content. In detail, 25kg of B. monnieri used to produce 1kg of standardized product (Synapsa) with 55%±5% active bacosides content. The other components in the extract such as alkaloid may enhance the bioavailability of the bacosides.

The manufacturer is selling the product (CDRI-08) in different names, i.e., Keenmind and Membac in New Zealand and Australia; Memo plus gold in Malaysia, Philippines, and Singapore; Memory perfect in France and Germany; and Synapsa in the USA. Two of the most popular patented extracts include Bacognize and Synapsa.

8.11. B. monnieri as a nutraceutical in commercial food products

B. monnieri is stated as a nootropic herb, which further enhances mental sharpness and memory acuity [109] and can be considered as functional food. B. monnieri contains carbohydrates, fat, proteins, and minerals, including calcium, phosphorus, and iron [110]. The first brand B. monnieri extract with GRAS status and so it could be used as one of the ingredients in bakery products, beverages, and candies at levels of 50mg per serving (https://naturalremedieshumanhealth.com/bacomind/GRAS). There are numerous products available in the market of India, which contains B. monnieri as a nutraceutical/functional food ingredient. It has used as a component in a number of various products including drinks, syrups, jellies, biscuits, and morning meal cereals [110]. The combinations of other herbs, i.e., Centella asiatica, Withania somnifera, Convolvulus pluricaulis, Tinospora cordifolia, Phyllanthus emblica along with B. monnieri are available in the market in the name of “Pitambari's Geni Health and Memory Enhancer Granules,” “Panchwati Health Prash,” “Ojasvita,” “Dabur Chyawan junior Malted Food Drink,” “Patanjali Brahmi sharbat,” and many more. These combinations are prescribed for increase in grasping power, blood proteins, sharpens memory power, and corrects nervous debility. B. monnieri is suggested as a product ingredient for the management of mental conditions including anxiousness, poor cognizance, inattention [111], and epilepsy.

Pharmacologically, the combination of active constituents is beneficial in mentally incompetent disorders and also in the management of neurological ailments. It is summarized that consumer trends are driving demand for ingredients supported by clinical research, quality, sustainability, clean labels, and robust adaptogenic assistances. This chapter only further exemplifies that traditional use with B. monnieri are well allied with consumer's demand in a product for cognitive vitality, emotional and mental well-being, and also restorative sleep (Fig. 8.3).

8.12. Future perspectives

A protocol should be assessed for drying and packaging of B. monnieri to retain the active constituents, i.e., triterpenoid glycosides content during storage all in a hygienic way. However, recently published study by Silpa et al. [112] showed the positive effects of drying processes, packaging material, and storage period on the bacosides content and microbiological quality. This plant (both wild and cultivated) is in consistent demand due to its well-known pharmacological, commercial, and ornamental use [113], which ultimately requires alternate for the production of the mass scale propagation method of this plant by in vitro multiplication procedures [114,115]. However, few tissue culture procedures have been previously reported for in vitro production of this plant. Some of the existing protocols propagated plant [116] without affecting secondary metabolite content and pharmacological effects, but for the mass production, the methods should be further improved.

Figure 8.3
Conclusive effect of Bacopa monnieri.

Acknowledgment

Authors are grateful to the Director, CSIR-Institute of Himalayan Bioresource Technology, Palampur, India, for support and encouragement. IHBT communication number is 4968.

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Chapter 9: Trigonella foenum graecum

Prajya Arya, Neha Kumari, Sajad Ahmad Wani, and Pradyuman Kumar
Islamic University of Science and Technology, Jammu & Kashmir, India

Abstract

Fenugreek also called as Trigonella foenum graecum, belong to Fabaceae family. It is one of the most common Indian spices that are particularly recognized for its culinary, medicinal, and aromatic properties. Fenugreek is more pronounced for its health-promising properties such as antiinflammatory, antiulcer, antidiabetic, and anticancer activities. Fenugreek is enriched with various chemical constituents that aids in enriching its nutraceutical properties. Its chemical constituents majorly composed of steroids, alkaloids, flavonoids, fibers, vitamin, and fatty acids. Fenugreek is enhancing human health by providing different bioactive compounds, antioxidant compounds, and hepatoprotective capacity. Fenugreek gel formation property aids in preventing antiulcer as studied with respect to some antiulcer drugs available like omeprazole. Fenugreek is one major kitchen ingredient that controls various metabolic diseases by its different nutraceutical properties. Based on the past evidences, the present review chapter aims to elaborate the nutraceutical properties of fenugreek.

Keywords

Fenugreek; Food value; Health beneficial ingredient; Nutraceutical properties; Spice

9.1. Introduction

Fenugreek is also known as Trigonella foenum graecum, is an annual grown herb that appertains to the family Leguminosae, well known from prehistoric times for its nutrition and medicinal properties. Fenugreek is a multiregional plant grown in India, the Mediterranean region, and the North American countries. The yield can be increased by applicable conduct of irrigation, cultivation, and harvesting. Seeds of fenugreek have promising medicinal properties such as lowering blood glucose level, anticancer, anorexia, hypocholesterolemic, gastric stimulant, used for lactation aid, and hepatoprotective effect. Fenugreek seeds are a great source of alkaloids, saponins, gum, fiber, flavonoid, and volatile compounds. The seed of fenugreek is bitter and used to enhance the flavor and color of the food [1]. Being high in fiber content, fenugreek has its usage as an adhesive, emulsifying agent, and stabilizer. Fenugreek is also used in modifying the texture of the food materials [2]. Studies have revealed seeds of fenugreek have numerous therapeutic effects, and most of the studies have indicated the advantageous effects to the antiinflammatory and antioxidant properties of fenugreek seeds [3].

The seeds and leaves of fenugreek are antiinflammatory, emollient, demulcent, expectorant, carminative, deobstruent, febrifuge, restorative, laxative, parasiticide, useful in burning sensation, and uterine. Since ancient times, fenugreek seeds have been used globally and are advantageous for muscles and bones, respiratory system, female reproductive system, gastrointestinal system, hepatic, cardiovascular system, and endocrinology. Studies have proved that fenugreek can be used to lessen cardiovascular risk and cholesterol. Fenugreek is considered as a good consolation for sore throats, constipation, a medicament in colon cancer, appropriate for acid reflux and skin infection. Fenugreek can remarkably lower down the menstrual discomfort and bring down the menopause symptoms. It can be used as an appetite suppressant and help in the prevention of weight gain [4].

Fenugreek has its usage in diverse industries such as nutraceuticals, food, and pharmaceutical industries as a traditional plant. Fenugreek perhaps can be a very efficient source of herbal drugs, and it could be used in the development of new drugs [5]. As further, fenugreek seed is a plentiful source of enormous essential compounds, and the components present in fenugreek seed play an efficient role in diseases prevention and control through regulating diverse biological activities [6]. From previous studies, it was observed that both traditional medicine and modern science with recently developed technologies and findings will assure the farming of medicinal herbs such as fenugreek and encourage feasibility in the long run [4]. The present review chapter elaborates the nutraceutical properties of fenugreek and utilization in various food products.

9.2. Morphology of fenugreek

Fenugreek is an upright, hairy aromatic annual spice of the bean family intently resembling large clover. The stem is generally reaching 30–60cm long. The height of the plant grows about three feet. The leaves of fenugreek are long stalked which is 5cm long stipule, triangular, narrow oval in shape [7,8]. The roots of fenugreek are mass of finger-like structure [7]. The sessile and axillary flower blossoms with white into yellowish white that is insect-pollinated and hermaphrodite. The pollen grains are oval, andand the ovary is deep greenish and glaucous in color [7,9,10]. Fenugreek flower has two to eight pods 15cm long which are brown and yellowish in color. The pod is sword-shaped each carrying 10–20 seeds. The plant diffuses spicy aromas that persevere on the hand after touching. The size of the seed is small, generally 5mm long. The seeds are hard, and the color varies from brownish yellow to dull yellow [11]. Fenugreek seed contains two cotyledons yellow in color surrounded by translucent endosperm. The germination time varies from 5–10 days in the soil. After 6–10 days of fenugreek germination, the first trifoliate leaf appears. The leaf development of the fenugreek is temperature-dependent [12]. Fenugreek seed attains maturity after four to seven months [13]. August to September during late summer is the ripening period and the midsummer from June to August is the flowering period of fenugreek seeds. Fenugreek is a dry land crop that develops well in a tropical climate with cool summer and mild winter. The botanical classification of fenugreek is depicted in Fig. 9.1.

Figure 9.1
Botanical classification of Trigonella foenum graecum.

9.3. Chemical constituents

Fenugreek contains several chemical constituents of numerous categorizations of secondary metabolite such as saponins, steroids, alkaloids, terpenes, flavonoids, phenolic acid derivatives, and amino acids [14]. There are some other chemical constituents present in fenugreek seeds which are presented in Table 9.1.

9.3.1. Steroidal compounds and saponin

Among the phytoconstituents present in fenugreek, saponin is the most abundant. Fenugreek contains 4%–8% saponins. The fenugreek leaves consist of seven saponins which are investigated as graecunins. These compounds are glycosides of diosgenin. Spirostanol and furostanol are the two classes of steroidal saponin. The majority of isolated saponin from fenugreek belongs to the furostanol class [14]. Diosgenin is the main sapogenin that is widely used in the pharmaceutical and nutraceutical industries. Its synthesis is mainly from cholesterol almost in all plants. The sapogenins after acid hydrolysis are diosgenin and yamogenin. Diosgenin is also extracted commercially [14,19,20]. Diosgenin is intricated in the production of human hormones. It is an estrogen precursor and used as a steroidal drug, and it also helps in managing menopause [19]. Diosgenin is used in the medication of hypercholesterolemia [21]. Due to the wide range of pharmacological activity, it is utilized as an antiinflammatory and anticancer agent [14].

9.3.2. Alkaloid

Trigonelline is the chemical compound that is the most substantial among the several pyridine alkaloids [16,22]. Trigonelline is neuroprotective and hypoglycemic biologically. Apart from this, trigonelline can activate estrogen receptors, so it is also classified as a phytoestrogen. The quantity of trigonelline is higher inside the seed than in pericarps, but the synthesis of trigonelline is much more in pericarps than in the seed [16]. At later stages, during germination, the synthesized trigonelline in the pericarps is transmitted to the seeds to be utilized [16]. The accumulated trigonelline is transformed to nicotinic acid, and later it is utilized for nicotinamide adenine dinucleotide synthesis [16]. Trigonelline is believed in reducing glycosuria in diabetes [19].

Table 9.1

Chemical constituent and bioactive compounds present in fenugreek seeds.

S. No | Chemical constituent | Bioactive compound | Reference
1 | Steroidal compounds and saponin | Graecunins, fenugrin B, trigofoenosides A–G, and fenugreekine | [15]
2 | Alkaloid | Neurin, Trimethylamine, Trigonelline, Choline, Carpaine, Gentianine, and Betain | [16]
3 | Flavonoid | Vitexin, Quercetin, rutin, and isovitexin | [17]
4 | Dietary fibers | Gum, detergent fiber, and neutral | [2]
5 | Vitamins and minerals | Vitamin A, B2, B1, C, nicotinic acid, niacin, carotene, folic acid, and calcium | [11]

6

Lipids, fatty acids, and esters

Triacylglycerols, monoacylglycerols, phosphatidylcholine, diacylglycerols, Phosphatidylethanolamine, free fatty acids, and phosphatidylinositol.

[1]

7

Amino acids

Soleucine, 4-Hydroxyisoleucine, Histidine,

Glutamic acid, Aspartic acid, Leucine, lysine, L-tryptophan, and Arginine

[18]

8 | Others | Coumarin, lipids, minerals, vitamins, 28% mucilage; 5% of a stronger swelling, 22% proteins, and bitter fixed oil | [1]

9.3.3. Flavonoid

A wide range of flavonoids is present in fenugreek seeds. Some of the important and beneficial flavonoids found in fenugreek are quercetin, vitexin, luteolin, and 7, 4-dimethoxyflavanones in the alcoholic extracts of the plant. Fenugreek has aglycones, kaempferol, naringenin, quercetin, and tricin in several part of the plant. Most of the flavonoids are found as complicated glycosides by conjugating with carbohydrates. The flavonol glycosides present in fenugreek are apigenin-6-c-glucoside (isovitexin), vitexin-7-O-glucoside (afroside), and quercetin-3-O-rhamnoside (quercitrin) [14].

9.3.4. Dietary fibers

Fenugreek is rich in fibers. Soluble and insoluble both two types of dietary fibers are present in fenugreek seeds. Fenugreek is one of the richest sources of dietary fiber which has 32% soluble fiber and 13% insoluble fiber. The fibers which are soluble break down in the aqueous medium of the gastrointestinal tract of the human, but the fibers which are not soluble do not digest by the enzyme. Soluble fiber plays a prominent role in enhancing glycemic control by inhibiting the absorption of glucose into the blood. Galactomannan present in the polysaccharides of the fenugreek cell wall provides energy for seeds in germination. The galactomannan contained in fenugreek is 13,000Da by molecular weight. The ratio of mannose to galactose monosaccharide is 1:1 in galactomannan present in fenugreek [14]. Galactomannan is a hemicellulosic polysaccharide that presents around 17%–50% of dry seed weight [23,24].

9.3.5. Vitamins and minerals

Fenugreek seeds are rich in vitamins. It consists of vitamin A, B2, B1, C, nicotinic acid, and niacin. Fenugreek leaf is rich in vitamin C. Every 100g of fenugreek fresh leaves contains 276mg of ascorbic acid content. Beta carotene, folic acid, and calcium are also present in fenugreek fresh leaves. Some of the essential inorganic elements are also present in the fenugreek plant. The major dietary elements present in the fenugreek plant are iron, potassium, magnesium, calcium, and sulfur [14].

9.3.6. Lipids, fatty acids, and esters

Fenugreek seeds have an enormous amount of mono and di-galactodiacylglycerols. Fenugreek contains almost 5.5%–7.5% lipids in total which comprises 5% glycolipids, 10% phospholipids, and 85% neutral lipids. Fatty acids present in the fenugreek seeds are linoleic acid, oleic acid, and linolenic acid. The important phospholipids are phosphatidyl, choline, and phosphatidylethanolamine. Fenugreek has N-acyl ethanolamines and oleamide, which help in relieving pain appetite-stimulating activity [14,25,26].

9.4. Roles of fenugreek as nutraceutical

So far, more than hundreds of phytochemicals have been identified and extracted from fenugreek seed, which includes saponins, polysaccharides, alkaloids, flavonoids, and polyphenols [27]. Fenugreek has an advantageous effect on cleansing the blood, it acts as a diaphoretic, which brings on the sweat, and helps in detoxing the body. Fenugreek has the ability for lymphatic cleansing activity; however, its vital role is to deluge the cells with nutrients and remove dead cells, toxic wastes, and trapped proteins from the body. Fenugreek also acts as a throat cleanser and mucous solvent, which may ease the urge to cough. Soaked fenugreek water assists in dissolving and softening, accumulating, and hardening the masses of cellular debris. Fenugreek is used in curing bronchial complaints, asthma, influenza, catarrh, sinusitis, constipation, laryngitis, pneumonia, sore throat, emphysema, hay fever, pleurisy, tuberculosis, and in relieving colds [1].

9.4.1. Anticancer property

Nowadays, one of the major causes of death in the worldwide is cancer. It can be believed that the total mortality that happened due to cancer could be decreased by adding up appropriate amounts of natural food to our diet [28]. Ref. [29] reviewed that nearly 50% of the medicines accessible in the market were extracted from plants sources and or chemically altered in the past 40 years.

The therapeutic medicines used in the treatment of cancer cause serious side effects. Active components that are plant-based have proved their prospective to be used as a safe and suitable substitute having remarkably surveyed anticancer affects. Investigations have revealed the effect of fenugreek seeds in opposition of cancer. It was affirmed that a component derived from fenugreek, protodioscin evinced an effect in inhibiting the development against HL60 cells by encouraging apoptotic moderation. In some other study, it was disclosed that the fenugreek extract stopped the mammary hyperplasia induced by 7, 12-dimethylbenz-anthracene [30]. The fenugreek extract exhibits cytotoxicity effect in vitro in opposition to the most of the various types of cancer cell lines in human-like HT29 cancer cell line, neuroblastoma, and IMR-32 [31]. The extracts of fenugreek have been inspected on the growth of MCF-7 cells and breast cancer cell line stating lower mitochondrial membrane potential and conversion of phosphatidylserine by the initial apoptotic changes and reduced cell viability after the expenditure of ethanol extract [32]. The anticancer property was investigated against prostate, pancreatic, and breast cancer; the findings revealed that fenugreek seeds are efficient in reducing the development of cancer cell lines [30]. Ref. [33] deduced that the diosgenin showed the promising application as an antitumor drug in the improvement of bioactivities. The diosgenin has been added up as a critical compound in modulating the biomembrane affinity, in the preparation of liposomes and liberation of hydrophilic cancer chemotherapeutic agent, cytarabine (Ara-C) from lipid vesicles. The steroid saponin compound in seeds of fenugreek, diosgenin displayed the elicitation of aberrant crypt foci emergence in F344 rats. It was observed that the multiplication of HT-29 colon cancer in human cells in a dose-reliant manner through the stimulation of caspase-2 led to apoptosis. It is documented that the seeds of fenugreek extract are eminently efficacious in the suppression of mammary hyperplasia and reducing its incidence in opposition of DMBA-induced breast cancer in vivo. Astonishingly, it revealed a specific cytotoxic effect in numerous cancer cell lines, yet no toxic effect was detected in normal cells. The usage of fenugreek extract in natural medicine might be an encouraging strategy as the chemically synthesized drug causes toxic effects to normal cells being a major disadvantage. The regulating biological processes within the cell such as gene expression, cell death, and cell division need induction of different signaling pathways by several enzyme complexes. Still, it needs the presence of diverse cofactors and coenzymes to get activated. Consequently, the usage of crude extract of fenugreek instead of active compounds alone might elevate the synergistic impression in between metabolites to persuade apoptosis through numerous signaling pathways [28].

9.4.2. Antidiabetic activity

Fenugreek seeds are helpful in reducing the blood sugar level and cause betterment in the insulin response [27]. Hypocholesterolemic is the occurrence of abnormally low levels of cholesterol in the blood or the abnormal reduced level of cholesterol level in the blood. The oral ingestion of the fenugreek seeds aqueous and methanolic extracts at a dose of 1g/kg resulted in a hypoglycemic effect in mice [34]. Fenugreek seed contains an enormous amount of soluble dietary fiber (galactomannan), trigonelline, diosgenin, flavone 4-hydroxyisoleucine, C-glycosides, and other ingredients which exhibited hypoglycemic activity in animals. Ref. [1] stated that the dietary fibers present in fenugreek seed blunt the glucose after the meal. 45.4% dietary fiber is present in fenugreek among which 32% insoluble and 13.3% soluble, and the gum is composed of mannose and galactose. The hypoglycemic effect of fenugreek seeds has been recorded in animals and humans with type 1 and type 2 diabetes mellitus [35]. Digestive enzymes can be inhibited by flavone C-glycosides, may decrease the preparation of modified glycation end products, and activate insulin signaling [36]. 4-hydroxyisoleucine present in fenugreek seeds stimulates the pancreas to secrete insulin. Ref. [37] studied the effect of fenugreek-integrated therapeutic food on blood sugar level with 24 noninsulin-dependent diabetes mellitus patients. After the predetermined time, it was observed that postprandial and fasting blood sugar levels were decreased remarkably by providing 30g product for about one month. Majorly, the fiber content of fenugreek seeds helped in the reduction of blood glucose level and could be a better source for diabetes management. Ref. [2] did a study on diabetes type II patients and found that the blood glucose level reduced among the patients by 25%.

The study was conducted for observing the effect of fenugreek on blood lipids with diabetic rats. In rats, diabetes was induced by streptozotocin and dispensed with oral intragastric intubation at low, middle, and high dose of fenugreek extract and metformin HCL for a duration of one-and-a-half months. After the completion of time duration, diabetic rats, which were treated with fenugreek extract revealed lower levels of triglycerides, higher high-density lipoprotein (HDL) cholesterol and total cholesterol in a dose-dependent manner [38].

9.4.3. Lactation aid

The most approved and utilized herbal galactagogue considered in the world is fenugreek. Breasts are altered sweat glands, and fenugreek seed has been found to stimulate sweat production, as it consists of a hormone precursor to hike milk formation. Many scientists have reported that fenugreek can enhance the milk supply of nursing mothers within 24–72 after taking the herb [39].

Ref. [40] reported an investigation for 6 years in which 1200 women were involved, and two to three capsules were administered (580mg or 610mg) three times per day can make most of the experimental subjects produce more milk. The effect of fenugreek seeds was observed on 21 lactating goats for 9 weeks and fat percentage and milk yield were the responses of this study. The fenugreek portion in the diet of goats was divided in percentage as (0%, 25%, and 50%) as group A, B, and C. Diets of all goats were kept same, and milk yield and fat percentage were recorded on the daily basis and weekly basis, respectively. Group B of fenugreek diet (25%) revealed the maximum percentage of milk yield and fat percentage as compared to group A and group C. Nonsignificant differences in globulin, plasma total protein, cholesterol, albumin, glucose, and total lipids are found among the three groups [41].

9.4.4. Immunological activity

An immunomodulator may be defined as an agent that escalates or diminishes the immune response, and this is known as an immunomodulatory effect. A study was conducted using fenugreek extract effect on stimulatory immunomodulatory. Fenugreek extract was divided in three doses (50, 100, and 200mg/kg of body weight) and were given to Swiss albino mice for 10 days. The immunomodulatory effect was observed in relative weight in thymus, body weight, cellularity of lymphoid organs, delayed type of hypersensitivity response, hemagglutination titer, lymph proliferation, phagocytosis, quantitative hemolysis assay, and a major increase in phagocytic capacity of macrophages and phagocytic index [42]. Saponins present in fenugreek are described as immune-stimulating agents. It can be proposed that some of the components may have mitogenic effects on the immunocompetent cells. Some of the compounds present in fenugreek also possess antioxidant properties, and they may induce the immune-stimulating result; several antioxidants have been reported to possess immunomodulatory properties.

9.4.5. Antiobesity activity

Some studies revealed that fenugreek seeds extract supplement is effective in decreasing body weight and adipose tissue weight. The portable mechanism is ascribed to flushing out the carbohydrates from the body before getting into the bloodstream which may result in weight loss. The high content of soluble fiber in fenugreek forms a gelatinous structure that can slow the digestion and absorption of food from the intestine. This develops a feeling of satiety [43]. Galactomannan from fenugreek seeds has a significant antiobesity function by the formation of a viscous gel in the intestine by the inhibition of glucose and lipid oxidation [44]. The fat accumulation of the fenugreek seeds was reduced by inducing a high-fat diet and by improving dyslipidemia in rats. The inhibition of fat digestion and absorption improved metabolism of glucose and fat and enhanced insulin sensitivity and reduced lipase and improved antioxidant capacity [45]. 4-hydroxyisoleucine from fenugreek seed extract can reduce plasma triglyceride levels in obese mice, which reduced increasing body weight [46].

Similar findings had been discussed in Zucker rats, which had obesity problems. The seeds of fenugreek were given to these rats. The liver of rats was experimented, which were obese, a reduction in tumor necrosis, and hike in the membrane receptor and tumor necrosis factor receptor 2. In supplementary study, it was revealed that the existence of fiber in fenugreek menacingly reduced the craving of famish and amplified in obese experimental units.

The dietary supplementation of fenugreek is manifested to have a promising effect on the reduction of weight for a limited period. When the fenugreek powder was provided to obese rats for 14 weeks, they observed the alteration in nutritional values, body measurements, and reduction in body weight was identified.

9.4.6. Antibacterial and antifungal activity

Fenugreek has antibacterial and antifungal activity; it has been recently shown. The different parts of the fenugreek plant such as the leaves, seeds, stems, and roots have possessed antifungal properties by making aqueous extracts of methanol, petroleum ether, ethyl acetate used against strains of fungus Fusarium, Rhizoctonia solani, Fusarium graminearum, Alternaria sp., and Pythium aphanidermatum. The potentiality of the antifungal property was displayed by all the extracts made by a different part of the fenugreek plant. Studies have shown that fenugreek can be used as a source of the biological compound in the improvement of a drug against the antifungal agent. In the recent past, Ref. [47] examined the antifungal activity against Microsporum gypseum and the results described that 100mL concentration of the ethanolic extract of the fenugreek seeds developed the highest inhibitory zone revealing 38.4% inhibition. In another study, it was found that fenugreek showed the highest actions against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus in honey samples.

Fenugreek seed extract prepared in methanol was allowed to expose against nematicide. Fenugreek extract causes destruction of 92% larvae of Meloidogyne javanica.

Fenugreek can be used in many food products against fungus and microbes. It has an affirmative effect on the body against harmful bacteria [30]. However, the researchers acknowledged the main antifungal activity was ascribed to the methanol fraction that suppressed the R. solani and Alternaria species extension. From the present study, it can be concluded that Trigonella foenum can be demonstrated as a useful source of the dynamic biological compound in the improvement of new drugs against fungus as an antifungal. Not long ago, Sudan and his team surveyed the fenugreek seed extracts antifungal activity against M. gypseum. The documented outcome illustrated that the fenugreek extract prepared in ethanol at 100μL concentration formed the utmost inhibitory zone (16.5mm) exhibiting 38.4% inhibition. The fenugreek seed potential is represented in Fig. 9.2 depicting various health-promising factors via therapeutic properties.

Figure 9.2
Therapeutic view of fenugreek seeds in controlling various health aspects.

9.4.7. Antioxidant property

Previous studies revealed that fenugreek seeds have antioxidant properties. Fenugreek seeds extract was prepared with different solvents such as methanol, dichloromethane, acetone, hexane, and ethyl acetate and had a good antioxidant capacity [48]. The seeds of fenugreek husk and cotyledon had the highest saponin and protein content [49]. It was indicated from the study that the separation of fenugreek seeds into husks and endosperm could have the advantage of process viability concerning prior selective fractionation of bioactive components for their effective isolation. Moreover, fenugreek seed extract regulated the activities of antioxidant enzymes that protected the elevation in lipid peroxidation. Consequently, it displayed an astonishing antioxidant potential in opposition to cisplatin-induced nephrotoxicity and hepatotoxicity. This may provide a way with the prevention of chemotherapy-induced toxicity. Additionally, it was suggested that the adding up of fenugreek seeds in diets would help in the reduction of the lipid peroxidation through the acceleration of decreased GPx, GST, SOD, and catalase as well.

The phenolic content of Aegle marmelos, fenugreek, and Coriander sativum was studied, and fenugreek had much higher content of flavonoid content as compared with A. marmelos and C. sativum. Further, in this study, A. marmelos produced better reducing power as compared with C. sativum and fenugreek. While fenugreek revealed much better free radical and superoxide scavenging activity as compared with Aegle marmelos and C. sativum [50].

9.4.8. Lowering and healing liver's hepatotoxicity and genotoxicity

A polysaccharide extract from fenugreek seeds significantly decreased and healed in a dose-dependent manner in the liver's hepatotoxicity and genotoxicity induced by thiamethoxam injuries. Ref. [51] conducted a study aimed to assess the preventive effect of a fenugreek-extracted polysaccharide against insecticide thiamethoxam-induced hepatotoxicity. The obtained data from this experiment showed influential antibacterial and antioxidant potentialities. On another move, in vivo experiment was conducted in adult female rats, which were divided into four groups. Insecticide thiamethoxam was administered 100mg/kg of the body weight. Again, fenugreek-extracted polysaccharide and insecticide thiamethoxam at two categorized doses of 100 and 200mg/kg of the bodyweight for about 30 days.

9.4.9. Anthelmintic activity

Fenugreek seeds have good anthelmintic properties as studied by Ref. [52] with alcoholic and water extract of fenugreek. Alcoholic-soluble fenugreek extract revealed better and progressive activity against anthelmintic as compared with water-soluble fenugreek extract. Soil can transmit helminthic infestation in a wide range in rural India, and fenugreek leaves being a cheap and easily available source can prove an alternative treatment for anthelmintic drugs. Fenugreek can be used to treat some parasitic infections in animals and humans. Ref. [52] conducted a study in vitro model and found that the aqueous extract fenugreek leaves paralyzed and led to the death of the worm. Ref. [53] studied in vitro anthelmintic activity of fenugreek seeds extract against Pheritima posthuma and found that the ethanol and the aqueous extract of fenugreek were used in the evaluation of the anthelmintic activity. The observed result of anthelmintic activity on earthworm P. prosthuma revealed the alcoholic extract and the aqueous extract of fenugreek have displayed paralysis against the death of earthworms when compared to similar concentration as the reference drug (albendazole). This study also concluded that an alcoholic extract of seeds of fenugreek has a potentiality against anthelmintic activity when compared with the conventionally used drug. It can be compared with standard drugs available in the market.

9.4.10. Antiulcer property

Fenugreek seeds aqueous extract and gel fraction proved to be good in treatment in gastric ulcer and gastritis [19]. The fenugreek soluble gel fraction is helpful in gastric lesion, and it produced much better results with omeprazole. Antisecretory and gastroprotective nature of fenugreek seeds, flavonoids, and polysaccharides revealed better results. There are a diverse number of products that could be used as antiulcerogenic, but the maximum number of the product users have some side effects which includes gynecomastia, arrhythmias, impotence, and hematopoietic changes [54]. Ref. [55] evaluated the effect of fenugreek seed extract in indomethacin-induced gastric ulcer models. The Albino Wister rats were categorized into five groups with each consisting of eight animals. Indomethacin 20mg/kg was used to induce gastric ulcers. The dose of 200mg/kg and 400mg/kg of fenugreek was administered to groups III, IV, respectively. Group V was administered with ranitidine 30mg/kg. The volume of gastric juice and ulcer index was evaluated. It was observed that group III displayed an eminent reduction in the ulcer index, acidity, and the volume of gastric juice was also decreased as compared to control group II. The extract of fenugreek seeds has both gastroprotective and antioxidant properties. Ref. [55] conducted a study and compared the effect of fenugreek to omeprazole on the ethanol-induced gastric ulcer. The aqueous extract and gel fraction extracted from the seed displayed an eminent ulcer protective effect. The fenugreek seeds showed a cytoprotective effect due to antisecretory action and the effect of mucosal glycoproteins. The seeds of fenugreek were claimed to prevent the hike in lipid peroxidation induced by ethanol. Previous studies have also disclosed that fenugreek-extracted gel fraction is more effective than omeprazole in preventing lesion formation. This proves that fenugreek seeds possess antiulcer properties.

9.4.11. Rheumatoid arthritis

A joint disorder attributed to inflammation, synovial proliferation, tissue destruction, and pain in one or more joints is arthritis. Interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Proinflammatory cytokines play a major role in increasing joint pain and stiffness. Ref. [56] conducted a study and observed that fenugreek has antiarthritic potentialities. Fenugreek mucilage has antiarthritic potential, which is helpful in inhibiting edema by suppressing inflammatory enzyme activities. The fenugreek mucilage played an important role in preventing arthritis in adjuvant tempted rats with arthritis. The fenugreek effect was stronger than that of standard drug, indomethacin, and the activity of inflammatory enzymes and mediators’ concentration was suppressed by using fenugreek mucilage in treatment of arthritis. Fenugreek supplements were given to patients suffering from inflammation in synovial fluid for bringing normalcy in synoviocytes. Based on the results obtained, it can be suggested that fenugreek mucilage possesses promising antiarthritic property, and it can be used as a therapeutic agent for arthritis.

Ref. [57] designed a study on Freund's adjuvant-induced arthritis in albino rats and investigated the profitable outcome of the ethanol extract of the fenugreek. In this study, they measured the paw volume of the albino rats on 12s were collected for the evaluation of the hemoglobin, white blood cells, erythrocyte sedimentation rate, interleukins (IL-1α, IL-1β, IL-2, and IL-6), red blood cells, and TNF-α. The cartilage tissue of the animals was isolated for the determination of superoxide dismutase, lipid peroxidation, and glutathione. Ingestion of fenugreek doses displayed a reduction in paw edema, and the body weight was restored. Fenugreek reduced the white blood count, and an erythrocyte sedimentation rate and a hike in red blood cells were observed. The study claimed that fenugreek can be used for antiinflammatory and antioxidant activities, which is the possible reason for the antiarthritic activity.

9.4.12. Fenugreek in asthma treatment

Asthma is one of the most commonly occurring inflammation of lung, especially in bronchial tubes by causing stiffness and hindrance in the air passage. Fenugreek played a crucial role in maintaining good lungs and their air exchange capacity. The polyphenol of fenugreek is rich in low molecular weight flavonoids that act as a key factor in controlling and treating asthma. From some of the literature review, it was observed that fenugreek extract was used in honey-based syrup to treat mild asthma and it is recommended as adjuvant therapy. Fenugreek and honey-based syrup aid in controlling and treating asthma [58]. The content of the flavonoid present in fenugreek seeds can hinder stimulation of mast cells and basophils that constrain lipid peroxidation and prevent nasal airways from the stress of oxidation. Quercetin flavonoid present in fenugreek seeds arrests the eosinophil cationic proteins and production of crystal which generate asthma pathogenesis. Fenugreek also has the capacity to repair the cell injury caused by oxidative mediators up to some extent. The flavonoids present in fenugreek can prevent the development of microcapillary tubule and cell migration [59]. Fenugreek mucilage helps in relieving asthmatic cough and brings comforts in normal and healthy secretion of the lungs. Ref. [60] conducted a study and observed that when fenugreek and honey syrup combination was utilized, it expedited the efficacy of the formula. When observed, fenugreek syrup revealed much better results as compared with honey syrup. Fenugreek can be used as a chest wall analgesic, lung tonic, and enhances breathing and lung secretion [58]. Fig. 9.3 depicts the various human health-promising aspects of fenugreek seeds.

9.5. Fenugreek as functional food

Utilization of fenugreek in the preparation of bread was observed to limit the blood glucose level; there was no significant change in texture, color, firmness, proximate composition, and flavor in fenugreek bread. The fenugreek bread was focused to reduce the insulin resistance and aided in enhancing fenugreek bread functional properties. It helps in the treatment of diabetic patients as well [61]. Ref. [62] conducted a study by incorporating fenugreek flour up to 10% in the formation of biscuits. The organoleptic assessment, baking quality, and color attributes of fenugreek biscuits were prepared by altering wheat flour from 10% to 20% of soaked and germinated flours. This proportion of biscuits makes it more acceptable while enriching health benefits. The addition of fenugreek seeds flour as raw, soaked, and germinated in biscuits efficiently reduced the low-density lipoprotein cholesterol, summate lipids and cholesterol. This study also observed some alterations in triglycerides serum HDL cholesterol and in reducing lipid in the cellular metabolism.

Figure 9.3
A pictorial representation of fenugreek seed health promising factors.

There are some other studies that proved the incorporation of fenugreek seed and leave the powder in the development of extruded snacks [63,64]. The extruded food products were enriched with colorful, functional, and physical properties with more acceptability and higher quality characteristics. The usage of debittered fenugreek polysaccharide for glycemic index, sensory quality characteristics, and physical parameters of fenugreek and chickpea rice-based extruded products were prepared and studied. This study revealed that 15% debittered fenugreek produced better results with more acceptability and less glycemic index [65].

In the recent development of the food industry, the focus has shifted toward producing more commonly consuming daily basis food products. The concept of utilizing various bioactive compounds makes it possible to deliver more nutrition to the end consumer. Fenugreek gum is composed of two aldohexoses comprised galactomannan are joined by α 1-4 glycosidic bond. This placement of bond prevents polymer chain from associating closely, which cause complete solubility in water at low temperature. The better rheological properties of fenugreek gum are imparted by galactose and mannose [66]. Ref. [67] studied the effect of incorporating fenugreek gum in the preparation of chapati (Indian unleavened flatbread). The physicochemical properties of fenugreek gum chapati were studied in respect of functional properties. The properties studied are swelling index, stability, water-holding capacity, oil-holding capacity, foaming, and emulsion capacity. The prepared chapati with fenugreek gum revealed better retention of functional properties of fenugreek gum. The prepared chapati had better texture, pliability, and softness even after keeping it for two days at room temperature. The utilization of fenugreek in ways in day-to-day life makes consumption more. It can be a possible way to utilize the health-promising benefits of fenugreek.

9.6. Conclusion

In the present chapter, an attempt has been made to explore the nutraceutical and functional food uses of fenugreek. Currently, herbs have fascinated more attention in medicine ascribed to the rising concern about health and safety. Fenugreek, a herb has not only the potential to serve as a medicinal herb and spices but also can protect, prevent, and treat a broad array of human diseases. The nutritional value of fenugreek and the presence of bioactive components are certainly appreciated by medical science. On account of presence of bioactive compounds and chemical constituents such as amino acids, alkaloids, and flavonoids, it can function as a good antioxidant agent. The fenugreek seeds have therapeutic properties such as antidiabetic, hypocholesterolemic, and serve as lactation aid. It influences immune functions, anticancer properties, antimicrobial, gastro and cardioprotective effects, antiarthritic, and serves as a strong antioxidant. Moreover, the clinical application of fenugreek is characterized by its manifold chemical composition, which proves it as a power plant to reduce the dependency on several synergistic drugs for disease current. Nevertheless, further research and investigation consisting of advantageous and well-planned clinical studies are required to strengthen these findings to produce a booming result for mankind.

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Chapter 10: Rosmarinus officinalis

Febin Pappachan, Amaya Suku, and Sreejith Mohanan
New Product Development, Plant Lipids Private Limited, Cochin, Kerala, India

Abstract

Rosemary (Rosmarinus officinalis L.) is one among the popular herbs since ancient time. Due to their excellent medicinal property, they were widely exploited all over the world in various therapeutic functions. Various extraction and purification techniques, in complement with different antioxidant assays revealed that they are packed with phenolic compounds, such as carnosic acid, carnosol, rosmarinic acid, ursolic acid, etc. Rosemary oils and extracts are hence popular for their exceptional antioxidant, antibacterial, as well as cognitive functions. As the results of which, they dominate in the list of well-known naturally sourced antioxidants in the food industry. Rosemary active compounds are hence being widely utilized in multiple food applications like in commercial frying oils, shelf-stable beverage drinks, snack foods, and meat and meat products. In several food application studies, rosemary extracts have proven effective on significantly reducing the level of oxidation, photo-oxidation, and lipid auto-oxidation. They also expressed significant effect in retarding color degradation and reduced rancidity notes by decreasing the level of oxidative reactions. While carnosic acid showed effective in oil/fat matrices such as frying oils and meat products, rosmarinic acid worked well in water-soluble matrices like beverage drinks especially in their color retention by inhibiting oxidation. The storage stability of rosemary-incorporated food products was also enhanced due to their potential antibacterial properties. In the case of health-promoting effects, rosemary phenolic compounds exhibited potential cognitive functions, along with ursolic acid acting upon the overall immunity boosting function of the body. The rosemary phenolics were also identified to have better performance in comparison with synthetic antioxidant such as butylated hydroxytoluene, butylated hydroxyl anisole), and tert-butyl hydroxyl quinone in some application studies, hence qualifying for their use as natural alternatives for similar antioxidant effect. Also, rosemary extract when in combination with other naturally occurring antioxidants such as ascorbic acid expressed synergistic effect with superior antioxidant properties.

Keywords

Carnosic acid; Rosemary oil; Rosmarinic acid; Rosmarinus officinalis L.; Ursolic acid

10.1. Introduction

Rosemary is an ancient herb and well known for its various medicinal properties. Scientific name of Rosemary is Rosmarinus officinalis L. (nowadays called as Salvia rosmarinus) and it belongs the family Lamiaceae (Labiatae) (Fig. 10.1). This herb, primarily belonged to Mediterranean regions, now turn out to be well-known and widespread all over the world. This evergreen shrub largely grown or cultivated at hilly regions of Morocco, Tunisia, Spain, Portugal, Turkey, Tunisia, Algeria, and India. Rosemary's oil and oleoresin have economic importance and used in various food applications. Main bioactives present in Rosemary are carnosic acid (CA), carnosol, rosmarinic acid (RSA), and ursolic acid (UA). Fig. 10.2 and Table 10.1 depict the structure of these bioactives and its specifications, respectively.

Rosemary contains about 1.5%–2.5% of essential oil, which can be recovered by steam distillation or hydrodistillation or microwave distillation or supercritical fluid extraction [1]. This oil is used as a flavoring agent and also used in medicinal and cosmetic formulations. Rosemary extract contains superior natural antioxidants, which is an approved food additive (E392) by the European Food Safety Authority (EFSA). Nowadays, rosemary extracts are used in various applications to replace synthetic antioxidants such as butylated hydroxyl anisole (BHA)—E320, tert-butyl hydroxyl quinone (TBHQ)—E319, and butylated hydroxytoluene (BHT)—E321. The quality specification required for rosemary extract is given in Table 10.2 and Table 10.3 as per E392 and JECFA (Joint FAO/WHO Expert Committee on Food Additives), respectively, (Commission Regulation (EU) No. 231/2012 and FAO JECFA Monograph 19, 2016).

Figure 10.1
Photograph of the herb Rosmarinus officinalis.

Figure 10.2
Important bioactive components in Rosmarinus officinalis.

Table 10.1

Specification of the major bioactives.

Name | Molecular formula | Molecular weight | XLogP3-AA | Solubility
Rosmarinic acid | C_18H_16O_8 | 360.3 | 2.4 | Water soluble
Carnosol | C_20H_26O_4 | 330.4 | 4.4 | Oil soluble
Carnosic acid | C_20H_28O_4 | 332.4 | 4.9 | Oil soluble
Ursolic acid | C_30H_48O_3 | 456.7 | 7.3 | Oil soluble

10.2. Rosemary oil and their roles in nutraceuticals and functional foods

Rosemary oil is considered to be safe and found application in different fields from the ancient times itself. The new regulatory conditions also agree with the safety profiles of rosemary oil. In 1965, the Fragrance and Essence Manufacturers Association of the USA have notified rosemary oil for Generally Recognized as Safe. It is seconded by the United States Food and Drug Administration (USFDA) by listing rosemary oil for food use. Also, the Council of Europe included rosemary oil under the category of substances, seasonings, and spices considered permissible for use, with a limitation of active principles in the end product [2,3]. Rosemary essential oil is widely used in aromatherapy and cosmetics due to its properties such as it helps to reduce stress levels, support healthy digestion, strengthen immunity, reputed to calm irritated skin, and support healthy hair growth (https://www.newdirectionsaromatics.com/blog/recipes/rejuvenate-yourself-with-rosemary oil.html). This reference gives details of the various formulations possible with rosemary oil. Table 10.4 shows the major components present in rosemary oil [4], which shows that α-pinene and camphene are the major components present in the oil. Rosemary oil has good antimicrobial activity and are mainly attributed to the presence of α-pinene and 1,8-cineole and thus used in packaging material to increase the shelf life of food products [5–7].

Table 10.2

Rosemary extract manufacturing regulations E392 (Extracts of rosemary).

Product details | Specification

Extracts of rosemary produced from dried rosemary leaves by acetone extraction

Assay: >10% total of carnosic acid and carnosol

Antioxidant/volatiles ratio: >15

Acetone: NMT 500ppm

Extracts of rosemary prepared by extraction of dried rosemary leaves by means of supercritical carbon dioxide

Assay: >13% total of carnosic acid and carnosol

Antioxidant/volatiles ratio: >15

Ethanol: NMT 2%

Extracts of rosemary prepared from a deodorized ethanolic extract of rosemary

Assay: >5% total of carnosic acid and carnosol

Antioxidant/volatiles ratio: >15

Ethanol: NMT 500ppm

Extracts of rosemary decolourized and deodorized, obtained by a two-step extraction using hexane and ethanol

Assay: >5% total of carnosic acid and carnosol

Antioxidant/volatiles ratio: >15
Hexane: NMT 25ppm
Ethanol: NMT 500pp

Key reference volatiles: Borneol, bornyl acetate, camphor, 1,8-cineole, verbenone

Arsenic: NMT 3ppm

Lead: NMT 2ppm

Table 10.3

Joint FAO/WHO expert committee on food additives (JECFAs) specifications.

Assay | Not less than 5% of the total carnosic acid and carnosol
Solubility | Insoluble in water; soluble in oil
Description | Beige to light brown powder

Antioxidants/reference volatiles ratio

Total % of carnosic acid and carnosol/total % of reference volatiles: (−) borneol, (−)-bornyl acetate, (−)-camphor, 1,8-cineole (eucalyptol), and verbenone: not less than 15

Loss on drying | Not more than 5%

Residual solvents

Acetone: not more than 50mg/kg

Ethanol: not more than 500mg/kg

Arsenic | Not more than 3mg/kg
Lead | Not more than 2mg/kg

Key reference volatiles: Borneol, bornyl acetate, camphor, 1,8-cineole, verbenone

Arsenic: NMT 3ppm

Lead: NMT 2ppm

Table 10.4

Rosemary oil gas chromatography profile.

Compound | Oil (wt %)
α-Pinene | 37.26
Camphene | 26.24
β-Pinene | 5.53
Sabinene | 5.05
Myrcene | 2.75
Limonene | 2.4
1,8-Cineole | 2.36
γ-Terpinene | 2.26
p-Cymene | 2.23
3-Carene | 1.9
Camphor | 1.72
Linalool | 1.42
Bornyl acetate | 1.24
β-Caryophyllene | 0.86
Terpinen-4-ol | 0.86
Verbenone | 0.83
Borneol | 0.6
Geraniol | 0.54
Neral | 0.35

10.2.1. Effect of carnosic acid on improving oil stability and deep frying

Deep frying of food items in fat is one of the renowned techniques for preparing food across the globe [8]. Deep-fried foods are famous among consumers due to its crispy texture, fried food flavor, golden brown color, and other appealing sensory attributes. Cafeterias and restaurants opt for this technique to quickly prepare shelf-stable convenience foods, potato chips, nuts, and frozen or refrigerated items such as meat, fish, French fries, vegetables, fish, etc., for sale in the retail market. Even though there is a growing concern among the consumers on the health impacts of deep-fried foods, its demand keeps on rising. This frying process involves immersing the food item in oil heated typically at a temperature extending from 165°C to 190°C. At these elevated temperature ranges, chemical reactions such as oxidation, polymerization, decomposition, and hydrolysis occur in the frying oil, resulting in off-flavour (rancidity), loss of nutrients, and various deteriorative shifts [9]. These may also result in the generation of toxic derivatives such as peroxides, epoxides, and aldehydes [10]. Vegetable oil rich in polyunsaturated fatty acids (PUFAs) such as high-oleic oils is found to exhibit high oxidative stability and are preferred over hydrogenated oils rich in saturated fatty acids [11].

On the contrary, costs of a majority of the high oleic oils are greater than other commonly used oils due to their limited production. As a result, strong antioxidants that inhibit oxidation and preserve the nutritional value of PUFA with high reactivity are being a major solution to this challenge. This can also help in encouraging the consumption of healthy, unsaturated, and unaltered oils for deep-frying operations in the food industry. Consequently, there is an increasing demand for strong antioxidants that suppress oxidation and protect the nutritional quality of highly reactive PUFAs as one of the major solutions to this problem and to facilitate the use of unaltered, healthy, unsaturated oils for frying in the food industry and in restaurants.

Regulatory norms were placed for the synthetic antioxidants by various regulatory bodies like the USFDA, EFSA, and Food and Agricultural Association (FAO) for application in bulk oils and fats (usually to a level less than 200ppm), considering their potential toxicological effects [12]. However, lower concentration levels permitted may not efficiently enhance the oxidative stability to PUFAs upon frying process. Hence, nontoxic natural antioxidants such as rosemary could have the potential to effectively protect frying oils from oxidation at higher concentrations.

10.2.2. Rosemary as a potential source for increasing oil stability

Several approaches have been introduced by several companies and researches all over the world to enhance the oil quality, improve its oxidative stability, and subsequent enhancement in its shelf life. Some of these methodologies include mixing of different oils for more stability, research on the use of new spices giving higher stability of oils, as well as the use of natural or synthetic antioxidants. Numerous researches focus on discovering superior sources of natural antioxidants, which could efficiently replace the commonly used synthetic alternatives. Basically, the antioxidant property of plant extracts is attributed to a mixture of various phenolic compounds present in them [13]. Hence, numerous plants were investigated for their antioxidant activity and on potential action in oils and fats. Currently, most of the investigation in this field focus on some of the extracts such as R. officinalis, sage (Salvia officinalis L.), tea, coffee, and vegetables especially onion and pepper [14]. The source which is most likely to be used industrially among these plant extracts is rosemary, as it exhibits substantial antioxidant properties even at elevated temperatures.

In bulk oil, the antioxidant effects of commercially available rosemary extracts and its active components such as carnosol, CA, and RSA have been widely researched (Table 10.5). CA was found to have a dose-dependent suppression of the generation of primary and secondary products of oxidation, as well as their ability to scavenge free radicals for oxidation [15,16]. On the major rosemary bioactives, CA had a stronger antioxidant property than tocopherols but was still inferior to TBHQ [16]. Caffeic acid also exhibits the property of effectively decreasing pecan oil oxidation, outperforming all natural and synthetic antioxidants tested, but falling short of TBHQ [17]. The CA is applied in a variety of food applications as defined in Table 10.6.

Rosemary oil extracts function well in a variety of edible oils. The effectiveness of rosemary extract with their commercial replacements synthetic antioxidants is higher and which is estimated to be superior [19]. There is a recent development of a synergistic, multicomponent antioxidant formulation comprising green tea catechins, ascorbyl palmitate, tocopherols, and rosemary extract for the preservation of industrial marine oil concentrates containing 300mg/g eicosapentaenoic acid and 200mg/g docosahexaenoic acid [35]. The validity and reproducibility of those results, however, are always questioned because of the complicated nature of herbal extracts. Furthermore, after adding these extracts to oil, the sensory characteristics of the oils like its flavor and color will invariably alter, making it undesirable. As a result, several studies have attempted to investigate the synergistic effect caused by a combination of different natural antioxidants in commercial oil applications.

Table 10.5

Application of rosemary extracts in different oils and fats.

Oils | Rosemary dosage levels | Conditions | Major findings | References

Palm oil

Oleoresin rosemary extract (0.4%)

Storage after heating to 60°C for six days and frying at 180°C, 10 frying six consecutive days, total 60 frying

Retarded the oil deterioration during six-day frying. The two antioxidants were proven to lower the rate of oxidation of oils during frying

[18]

Refined sunflower oil

200ppm of rosemary extract having carnosic acid concentration 24.9%, 60.5%, 98.3% (w/w) compared with 200ppm of BHA, BHT and TBHQ

60°C for 21 days under darkness

Positive correlation between carnosic acid concentration and the oxidative stability of sunflower oil. 98.3% carnosic acid exhibited more effectiveness than BHA and BHT and less effective than TBHQ

[19]

Refined sunflower oil | 1000ppm rosemary extract | Thermal oxidation conditions (180°C) | Extended the oxidative stability from 7.52 to 13.5 hours | [20]

Refined soybean oil, rice bran oil, and cottonseed Oil

400ppm of rosemary extract andand compared

With 50:50 mixtures of BHA andand BHT (200ppm)

Accelerated oxidation condition

Rosemary extract exhibited much better chemical stability, including higher induction period, antioxidant capacity, total phenolic content, and lower peroxide value

[21]

Virgin olive oil | Carnosic acid (100 and 1000ppm) | Accelerated oxidation conditions at 60°C | Dose-dependent inhibition in the formation of primary and secondary oxidation products | [15]

Tocopherol-stripped corn oil

Rosemary powder and powder of black

Pepper, ginger, turmeric, and oregano at 0.5% (w/v) and BHT (100ppm)

Accelerated oxidation conditions

Rosemary powder was found to be the most effective one among all the natural ground herbs and BHT

[22]

Table 10.6

Application of rosemary extract in meat-based food products.

Meat product | Rosemary dosage levels | Shelf life study conditions | Observed effect on self-stability | References

Raw and cooked chicken patties

Extract (300 and 350ppm)

4°C for seven days

Decrease in lipid and protein oxidation, redness loss (raw), and rancidity odor (cooked); increased sensory score of color (cooked) −12

[23]

Raw beef patties | Extract (0.1%; 3%, 6%, and 10% of carnosic acid in the extract) | 4°C for 90 days | Reduced peroxide and thiobarbituric acid–reactive substances (TBARSss) values, and loss of redness - | [24]
Raw pork patties | Extract (0.1%) | 4°C for 15 days | Reduced loss of redness, metmyoglobin formation, conjugated dienes, and peroxide and TBARS values | [25]
Raw pork burger | Extract (0.1%, 0.2%, and 0.3%) | 4°C for 10 days | Reduced lipid and protein oxidation and redness loss (0.2%) | [26]
Raw chicken burgers | Extract (0.02%) | 4°C for 21 days | Reduced lipid oxidation | [27]
Raw beef burgers | Extract (28mL/kg) | −18°C for 60 days | Reduced metmyoglobin formation, lipid oxidation, and redness; no effect in sensory properties | [28]
Raw mixed meat burger | Extracts (0.015%, 0.03%, and 0.06%) | −12°C for 120 days | Low lipid oxidation and loss of redness; high acceptance among panellists | [29]
Cooked cured chicken sausage | Essential oil (5%) | Final product | Lower redness and lipid oxidation; no effect in sensory attributes | [30]
Cooked and smoked pork sausage | Extract (0.1%, 0.2%, 0.3%, 0.4%, and 0.5%) | Stored at 4°C for 20 days | Increased redness, reduced lipid oxidation, and VBNa (0.5%) | [31]
Table Continued

Meat product | Rosemary dosage levels | Shelf life study conditions | Observed effect on self-stability | References
Cooked cured beef sausage | Extract (250mg/kg) | Stored at 4°C for 25 days | Lower peroxide value and TBARS values | [32]
Chicken nuggets | Extract (150ppm of carnosic acid and carnosol) | Stored at −18°C for nine months | Reduced lipid oxidation; no effect in color, and rancidity odor and taste | [33]
Fermented pork sausages | Extract (0.48%; 100g/kg rosemary extract in the oil phase) | Stored at 7°C and 20°C for 49 days | Reduced POVc, TBARS and hexanal content, redness loss, and rancidity (sensory analysis) | [34]

10.2.3. Effect of carnosic acid on improving shelf life and color of meat

The quality of meat and meat products degrades predominantly due to the occurrence of oxidative unbalance and free radical generation [36]. Usually, the oxidative reactions in meat products initialize due to its exposure to oxygen present in the atmosphere, endogenous enzymes, transition metals, ultraviolet radiation, and the presence of free radicals [37]. The lack of appropriate preventive measures, leading to advancement of oxidative reactions in oxidation-prone components like myoglobin and lipids, leads to detrimental effects on the quality of the meat products during processing and storage. These effects could indeed lead to lower acceptance by the consumers due to the development of off-odor and degradation of color [38].

Natural or synthetic antioxidants are normally utilized for delaying the quality deterioration due to oxidative reactions in meat products [39]. Even though synthetic antioxidants are extensively applied in meat products, consumer distresses are increasing on its potential effects in health, leading to a shift toward the manufacture of clean-label products. At present, there are a wide variety of natural antioxidants available in the market. However, rosemary gained attention because of its remarkable antioxidant activity, owing to the presence of phenolic compounds (CA, carnosol, α-pinene, 1,8-cineole, and camphor) and appealing sensory characteristics [40]. The rosemary phenolics have been utilized in a variety of applications in meat-based foods (Table 10.6) such as raw or cooked meat, burger patties, nuggets, etc., These meat-based products are tasty and famous among the people; however, its quality is affected by oxidation [41].

During 90 days at 4°C, a comparable experiment in raw beef patties containing rosemary extract (3%, 6%, and 10% CA) and oleoresin of paprika revealed appreciably minor levels of peroxides (primary oxidation products) and thiobarbituric acid–reactive substances (TBARSs) than those prepared without any antioxidants [24]. The red color of the beef patties also remained unchanged during storage. There is a potential application of BHA and rosemary on controlling paprika discoloration. After 15 days, samples containing BHA had completely discolored, whereas the highest concentration of extracts retained the vivid and unique red color of paprika oleoresin.

The application of rosemary antioxidants in combination with different packaging technologies has generated promising results in terms of improving patties' oxidative stability and used rosemary extract in vacuum-sealed raw pork patties to test this method. In the case of vacuum-packed pork patties with rosemary extract, the development of metmyoglobin was reduced, and a considerable retention of redness was observed [25]. There was a subsequent delay and decreased intensity in the generation of conjugated dienes and peroxides (primary oxidation products) in the control samples without rosemary when it came to lipid oxidation. Rosemary extract also inhibited the production of secondary oxidation products (as assessed by the TBARS method).

The antioxidant effect of rosemary has also been documented in burgers, especially in patties combined with other seasonings and essential food additives. For example, while studying the influence of different concentrations of rosemary extract on raw pork burger preservation, 0.2% concentration exhibited better results in controlling the oxidation of protein and lipid under storage conditions [26]. Furthermore, the decrease of redness was minimized as a result of this treatment. A related study found decreased levels of lipid oxidation in raw chicken burgers stored for 21 days at 4°C compared to samples without antioxidants (control samples) [27]. A burger made of beef, chicken, and turkey meat had a similar result in a related experiment [29]. During frozen conditions of −12°C, all the studied concentration levels of rosemary extract could effectively decrease the color and lipids degradation due to oxidation for a period of 120 days. Furthermore, a sensory study demonstrated that higher concentration level of rosemary extract in this burger created a sensory experience of “freshness,” which enhanced panellist acceptability. During the frozen storage of raw beef patties, rosemary extract had a comparable protective effect. However, there was considerable reduction in redness [28].

Sausages are a category of meat products made from a mixture of animal fat and ground meat, along with seasoning and required additives (nitrate/nitrite) for preservation that are treated to achieve specific features [30]. The individual standards assigned to these products by the consumers, as well as the broad range of ingredients and respective processing conditions, are attributed to the widespread consumption and production of this category of meat products. Because these goods are produced under settings that promote oxidation like processes such as meat comminution, persistent exposure to atmospheric oxygen, and thermal processing, antioxidant-rich additives are commonly included. As a result, oxidation might occur, resulting in the creation of volatile products and sensory-active chemicals, which are responsible for the meat's expected odor and flavor [42].

In most of the cases, nitrite (incorporated in the form of sodium salt or formed during the fermentation process) plays a significant role in preventing oxidative degradation of quality [43]. However, discovering a suitable replacement for nitrite is currently one of the serious concerns in the meat industry. The difficulty lies in finding alternatives having equivalent or better capability to prevent oxidation and its related products, while simultaneously assisting in physicochemical and microbiological stability and safety [44].

Rosemary extracts as well as essential oil, on the other hand, because of their high antioxidant capacity, have been utilized as a natural source of antioxidant to increase sausage stability to prevalent oxidation. For example, the rosemary essential oil inhibited the development of lipid oxidation products in a cured cooked chicken sausage when in comparison with a nitrite salt control [30]. The redness of the rosemary and control sausages was likewise reduced, although there was no difference in sensory qualities.

The protective characteristics of rosemary extracts in cooked and smoked pork sausages stored in refrigerated conditions (4°C) and during 20 days of storage showed that rosemary extract inhibited the oxidation of lipids and related products as well as volatile basic nitrogen (VBN) in concentration-dependent manner (primarily 0.5% treatment) [31]. The intensity of the redness grew as the rosemary content increased and remained constant throughout the storage time. Similarly, over a storage at 4°C for 25 days, rosemary extract lowered the primary oxidation products (peroxides) and concentration levels of TBARS in cured and cooked beef sausage.

Nuggets are type of meat products which are typically deep fried in unsaturated vegetable oils. This process of deep frying leads to the absorption of oil in them and hence favoring oxidative processes while stored under frozen conditions. The rosemary extract usage can help nuggets to maintain their oxidative stability. Also, natural extracts in three different forms, i.e., liquid ethanol, liquid acetone, and powder acetone, lowered the oxidation of lipids following a storage period of nine months during freezing conditions. However, there were no differences in color, rancid odor, or flavor between rosemary and control nuggets [33].

10.2.4. Effect of carnosic acid on stabilizing carotenoids like paprika (evidences)

Paprika, dehydrated and crushed fruit of some pepper variety (Capsicum annuum L), due to its high carotenoid pigment concentration, is one of the most extensively utilized colorants in the food industry [45]. The main trait that determines the quality of paprika is its bright red color, which is determined by the total carotenoid content present in the fruit [46]. The total carotenoid content could vary widely depending on the species and other external factors such as maturity, climate, and cultivation circumstances [47]. Since carotenoids are easily oxidized, major color loss in spice paprika begins during processing and storage after harvest [48]. Furthermore, multiple studies have proved the impact of technological aspects [49,50]. During red pepper ripening, processing, and storage, the quantity of endogenous antioxidants has been found to be substantially correlated with color stability [48]. The quality of paprika deteriorates over storage, as it does with many foods. Its oxidation by free oxygen causes loss in color, which is initiated by the degradation of vitamins such as vitamin C and E and later continues with the destruction of red and yellow carotenoids [51]. At the same time, extrinsic elements, including physical characteristics such as temperature, light, humidity, etc., or chemical characteristics such as enzymes, peroxides, presence of metal ions, etc., could influence this oxidation [52].

Several researches have concentrated on the loss of color of paprika at various temperatures [52]. However, despite being largely accepted and popular, the pepper types that have been examined are rapidly being superseded with newer and other effective varieties obtained through techniques of plant improvement, giving peppers with intense and superior color profile [53]. Certainly, enormous research has focused in the color degradation in paprika obtained from these novel types, as well as the effect of temperature and humidity combination on their deterioration [54]. Furthermore, paprika powder is a color-enhancing component in a variety of dehydrated products, such as soups, sauce powders, and seasoning mixes, where increased color retention is a fair expressed or underlying need [55].

Food preservation has seen significant progress, particularly in the avoidance of rancidity in oils and fats, as well as the prevention of discoloration. Antioxidants have extended the half-life of a wide variety of processed foods, resulting in the reduction of unpleasant taste and odor development [56]. Up until now, synthetic antioxidants such as BHT, BHA, TBHQ, and gallates have been employed mostly [57]. However, these compounds have several drawbacks, particularly in terms of their potential degradation at elevated temperatures and volatility. Moreover, as people's knowledge of the potential risks of synthetic food additives has grown, several studies have been conducted to establish their toxicity. Many nations have banned the use of such antioxidants since they can harm the enzymes of lungs as well as liver, and as a result, the consumers’ interest has increased in natural antioxidants such as tocopherols, ascorbic acid, and R. officinalis extract.

The phenolic and flavonoid components and phenolic diterpenes in R. officinalis and its extracts possess antioxidant properties [58]. Yet, the major antioxidant activity is due to the CA and carnosol [59]. The incorporation of rosemary extract to the paprika salami improved the color and reduced the oxidation of lipids in meat as well as lard matrix. The reaction kinetic calculations in a research revealed that adding rosemary extract at 1% enhanced the color strength and stability of spiced paprika powders treated with heat [60].

10.3. Effect of rosmarinic acid in beverage drinks

10.3.1. Effect on shelf life extension

Functional beverages, comprising drinks enhanced with vitamins and minerals, juices, energy drinks, sports drinks, wellness drinks, and nutraceuticals, are a rapidly developing segment of the industry. Majority of functional drinks are produced with a motive to provide a health benefit or to deliver a specific medical need such as improving heart health, improving digestion and immunity, and energy enhancement [61]. The target market for these functional ingredients is wide, and the products are often made to adapt to the needs of a particular target group. These target groups may be based on parameters such as by gender, age, particularly emphasizing on women, elderly, and children. Normally, the lower pH range of soft drinks is expected to prevent bacterial deterioration. However, fungal growth can take place in the medium and affect the storage stability of the product. The primary contaminants in these types of beverages are yeast, owing to their ability to multiply and grow even in beverage drinks with high levels of carbonation, as well as lower pH ranges. As most of the common preservatives used in beverage drinks are derived from weak acids such as sorbic and benzoic acid, the growth may not be restricted [62]. Hence, the use of antimicrobial peptides of natural origin takes up the possibility to preserve food without altering its quality.

During the past decades, soft drinks emerged to be one among the daily consuming products in the diet. But, nowadays, soft drinks like beverage products are greatly incriminated because of the higher concentration of sugars and preservatives. This led to a decrease in their consumption in recent days [63]. Basically, soft drink quality is attributed to their low pH, antimicrobial and antioxidant additives, and the thermal processing (pasteurization) [64]. The major antioxidant material added to beverage drinks is ascorbic acid, which has the ability to inhibit oxidation of inherent components and quality enhancement. Presently there exists a mounting demand by consumers all over the world for “green label” foods, with natural additives, presenting properties to replace the artificial ones in terms of their antimicrobial as well as antioxidant properties [65]. Soft drinks and beverages have become a staple in the last few decades and fall under the group of daily consuming products for the majority of the population. However, consumption of products like soft drinks and beverages had decreased due to elevated levels of additives and sweeteners in them [63]. Therefore, the demand for natural preservatives with antioxidant and antimicrobial properties originates the need for natural sources to preserve the beverage drinks increased.

Normally, chemical preservatives are utilized to improve the microbiological stability of soft drinks and other beverages. The type of chemical preservatives to be utilized depends on the physical and chemical attributes of the beverage as well as the preservative. The types of preservatives to be used, if any, to prevent microbial development are determined by the pH, vitamins present, type of packaging, and storage settings. In Europe, the use of benzoates (E210–213), sorbates (E200–203), and dimethyl dicarbonate (E242) is approved in the case of ready-to-drink beverages. However, there is an increasing concern in using such chemical preservatives in beverage drinks, owing to their degrading effect on human health. Hence, there is an increasing demand for natural compounds with potential antioxidant and antimicrobial properties. In this need to find natural ingredients with antioxidant and antibacterial capacities, rosemary has been considered remarkable by many researchers [66,67]. The minimum inhibitory concentration (MIC) and minimum bactericide concentration of commercially available rosemary extracts compared to gram-positive bacteria like Bacillus and Staphylococcus and gram-negative ones like Campylobacter and Salmonella were investigated [68]. The antibacterial and antioxidant properties of the extracts were determined by the chemical composition and their concentration of the phenolic compounds, according to these scientists. Gram-positive bacteria were more susceptible to oil-soluble extracts compared to gram-negative bacteria, containing CA being the prominent phenolic component. Because rosemary extracts have antibacterial action toward Shigella sonnei, Salmonella typhimurium, and Listeria monocytogenes. Hence, RSA is a powerful water-soluble polyphenol from rosemary with good antimicrobial and antioxidant properties.

10.3.2. Effect on color stability of beverages

In consideration to creating public's expectations about the anticipated taste and flavor of a foodstuff or a beverage, color is a vital product-intrinsic sensory indicator. Several experts currently believe that our taste and flavor experiences are shaped on the basis of the expectations we form prior to tasting, particularly at times we may scrutinize a beverage drink visually prior to buying it or deciding whether to sample it [69]. The acquisition of food and drink is tightly tied to the brain and visual system development, and recent neuroscience reveals that the view of attractive food or drink is a dominant signal for the brain cells, particularly in the case if the brain is of a hungry individual [70].

The usage of colorings in soft drinks basically serves to several significant purposes such as:

- 1. making the product more visually appealing;

- 2. assisting in the correction of natural color variations or changes during processing or storage; and

- 3. assisting in preserving the distinctive qualities of the drinks.

The three basic categories of food colors include natural colors, artificial colors, and caramels. In this, natural colorings are subdivided into two categories, which could be taken from vegetables, fruits, or plants. These include:

- 1. yellow-orange carotenoids attained commercially from a variety of fruits and vegetables;

- 2. brilliant red-purple anthocyanins found commercially from a variety of fruits and vegetables.

Antioxidant properties of natural colorings also added to the soft drinks’ quality [71]. Recently in both the markets of the European Union as well as United States, the usage of artificial colors has been reduced by the sellers and manufacturers, owing to growing consumer aspiration toward natural colorants. Antioxidants are added to foods and beverages to prevent degradation and extend their shelf life. However, each meal or drink's appearance and attributes, such as color and sensory properties, must be preserved [72]. They must also be simple to use and cost-effective. The safety of food additives is confirmed by regulatory bodies, such as the EFSA or the USFDA, using conventional techniques to validate the use of compounds and quantities [73].

In recent years, consumers have shown a rising inclination toward phenolic natural antioxidants compared to artificial antioxidants, owing to a rising need for functional beverages and foodstuffs containing natural ingredients which retains the nutritional characteristics as well as flavor profile [74]. Furthermore, this tendency stems due to the present interest for natural antioxidants in the stability of food, as well as the restrictions placed on the use of synthetic phenolic antioxidants by the responsible authorities [75]. Researchers have paid the most attention to rosemary as promising natural sources of antioxidants. CA, carnosol, and RSA are the phenolic compounds, primarily accounting for rosemary extract's antioxidant properties. Rosemary is rich in water-soluble phenolic acid called RSA, which is one among the most potent antioxidant in simple phenolic acids [76].

10.4. Effect of carnosic acid and rosmarinic acid on cognition

One of the main health issues that many people often face during ageing is the reduction of their cognition abilities which is often linked with issues such as poor reasoning, loss of memory, perceptual speed, and language. When these issues start creating a prolonged effect with a reduction in functional activities of brain, its commonly linked to the term dementia and Alzheimer’s disease (AD, the most associated form of Dementia). By 2050, USA is likely to reach a million case of this disease per year [77]. An increasing demand to enhance the cognition improvement and to delay the occurrence of dementia has been in existence all long. Since a cure against these problems exists, public health priority started focusing on preventing cognition problems. Consuming nutritional foods available in nature, supplementation with nutritional components, cognitive as well as social activity, physical exercise are some of the identified available options for AD prevention [78].

CA and RSA, powerful antioxidants from rosemary, have been investigated [79]. In vivo studies as well as in vitro studies have showed the neuroprotective action; CA and RSA act to scavenge reactive nitrogen species [80] and guard neuroblastoma cells from oxidative stress induced by hydrogen peroxide [81]. RSA, a neuroprotective agent, helps in decreasing cell destruction and disorders caused by free radicals and several other aspects [82]. Memory impairments may be mediated by daily consumption of RSA. Protective nature of RSA against neurotoxicity of Aβ25–35, which may be due to the scavenging property of peroxynitrite [83]. Diterepenes reduce memory disorders linked with Aβ neurotoxicity in AD and help in the delaying the initiation of the illness [84]. It is also reported on the antiangiogenic effects of CA [85]. The mechanism about the inhibition of angiogenesis is yet not clear, but its antioxidants activity plays a crucial role in serving as an antiangiogenic.

10.5. Effect of ursolic acid

10.5.1. Antimicrobial effect

UA and its by-products have been found to limit the development of bacteria, particularly gram-positive and gram-negative, and fungi, and UA has been proven to restrict the activity of elastase enzyme, the growth of bacteria, and the formation of biofilm [86,87]. There is a discovered synergistic effect of UA and two semisynthetic products, as well as the aminoglycoside antibiotics such as amikacin, kanamycin, neomycin, and gentamicin, against 12 different pathogenic bacterial strains, with MICs in the range of 32 to 526g/mL [88]. UA is a major constituent extract that inhibits the growth of some food-related bacteria and yeast [89], protozoa (Trypanosoma cruzi) [90], and promotes an antiinflammatory response during Leishmania infection. Ref. [91] recently examined UA's antibacterial, antifungal, antiparasitic, and antiviral properties.

In the case of antimicrobial and antiviral works formulated by using natural extracts, pure UA, or its derivatives, UA is often used as an alternative antibiotic against Salmonella enterica [92], in all stages of agricultural product handling, and with antihepatitis C virus effect [93]. Microbial infections can be prevented and treated using a combination of terpenoids and antimicrobial medicines [94]. The use of UA in blend with penicillin against drug-resistant Staphylococcus aureus resulted in in vitro synergistic effect and superior antibacterial characteristics than either component alone [95]. Tobacco mosaic virus can be controlled with a plant virus restricting the chemical composition comprising glycyrrhizin and UA that is harmless for both humans as well as the environment [96]. Bacteriostatic, tumor-resistance, and antiinflammatory properties were found in rosemary extracts containing more than 90% UA [97].

10.5.2. Immunity-enhancing effect

Immunomodulatory activity is a term used to describe the action of a substance on the immune in Balb/c type of mice, by the administration of UA intraperitoneal way improved the total white blood cell count, cellularity of the bone marrow, and increased alpha-esterase positive cells, as well as hindered hypersensitivity activities inside the body. The treatment using various triterpenoids in grouping with antigen also increased explicit antibody titer and total number of plaque-forming cells within the spleen [98]. The treatment of UA intraperitoneally to animals with metastatic tumors boosted inherent killer cell activity as well as complement-mediated antibody-dependent cytotoxicity [99]. While considering type 1 diabetic in the case of mice fed with a high-fat diet (HFD), UA had immunomodulatory effects [100].

10.5.3. Anticancerous activity of UA

UA has proven characteristics as a powerful anticancer agent that works both in vitro as well as in vivo. In both rats and humans, numerous researches have looked into the positive impact of UA upon metabolism of cancer cells. Inhibiting the process of carcinogenesis [101] and cancer cell proliferation [102], regulation of apoptosis [103], prevention of cell cycle arrest [104], and activation of autophagy [105] are reported to be the mechanisms behind its anticancer activity.

10.5.4. Antiobesity/Antidiabetic activity of UA

In vivo and in vitro, the main actions of UA are suppression of pancreatic-amylase activity and lowering of blood glucose levels [106]. In a study that looked into the effects of UA on regulating the body weight as well as glucose tolerance in metabolic syndrome patients who took 150mg of UA before breakfast for 12 weeks, there was a decreased patients' body weight, body mass index, waist circumference, and blood glucose levels (fasting), suggesting that their insulin sensitivity has enhanced drastically [107]. A study also found that 0.5% UA-supplemented meal in a HFD-induced obese mice, expressed reduced body weight, free fatty acids, and oxidation over a period six weeks of treatment with uncoupling protein 3/AMP-activated protein kinase-dependent pathways [108].

10.5.5. Cardiovascular disease and effect of UA

Cardiovascular disease (CVD) is the leading cause of morbidity and mortality all around the world. Coronary artery disease, heart failure, myocardial infarction, stroke, atherosclerosis, peripheral artery disease, hypertension, and cardiomyopathy are all included in the CVD category [109]. The therapy with UA (40mg/kg) is related with a lower heart rate, indicating that CVD risk is reduced both in vitro and in vivo [110]. After seven days of therapy, it was found that UA (60mg/kg) lowers lipid peroxide levels by scavenging free radicals, improves lipid profiles, and lowers blood levels of membrane-bound proteins [111]. In rats, UA helps restore cardioprotective enzyme activity to normal levels, suggesting that it may protect against myocardial ischemia.

10.5.6. Brain function and effect of UA

Oxidative stress as well as excitotoxicity are known to cause mild to severe nervous system disorders [112]. UA suppresses the effects of oxidative stress and levels of excitotoxicity [113], signifying a potential application to protect against oxidative stress and excitotoxicity-related brain disorders. UA also inhibits apoptotic signaling and has antiinflammatory properties affecting the brain [114].

10.5.7. Liver diseases and effect of UA

UA has been shown to protect against a variety of liver illnesses, including fatty liver disease [115], carcinoma [116], liver fibrosis [117], and liver cancer [118] in numerous investigations.

10.5.8. Improving exercise activity and effect of UA

In vitro as well as in vivo studies have recently revealed that UA enhances exercise capacity via several molecular pathways. Nutrient supplements with UA improve exercise capacity while also lowering resting heart rate [119]. The expression of sirtuin-1 and peroxisome proliferator-activated receptor-coactivator-1 in the skeletal muscles of old rodents was shown to be increased after intraperitoneal treatment with UA for a period of seven days [120], indicating that UA may improve physical performance.

10.6. Conclusion and future perspectives

Rosemary is one of the superior herbs rich in powerful phenolic compounds such as CA, carnosol, RSA, and UA, which exhibits high performance against oxidative reactions in both chemical assays in vitro as well as in real food matrix applications. Rosemary extracts/essential oils are powerful antioxidant and antibacterial agent, with a great choice to be used as a natural food additive for preservation. At the same time, they could also impart to the flavor profile of the food products and hence could be used as a flavoring agent. Besides these two actions of preservation as well as flavoring, they could also contribute to the consumer health-promoting functions, proving their use as a promising plant source for food application (oils/fats, meat and meat products, beverages, and colored food products), especially in functional foods and nutraceutical industry. Combination of rosemary extract with other natural ingredients could lead synergistic effect imparting better performance. The phenolic compounds also exhibit cognitive as well as immunity enhancement function, especially UA. Rosemary essential oils could be utilized to replace the synthetic food additives in the current market, without compromising its antioxidant capacity. Also, in the future with improved regulatory norms, they could widen their use as a natural antioxidant in different food and nutraceutical applications.

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Further Reading

[1].
Commission Regulation (EU) No 231/2012 E392 Extracts of Rosemary, (02012R0231- EN-02.07.2020-022.001, Page 117–119.

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Chapter 11: Phytochemicals and therapeutic potential of Punica granatum L

Bhupinder Kumar
[1]
, Shah Alam Khan
[2]
, and Md Jawaid Akhtar
[2]

1
Department of Pharmaceutical Chemistry, ISF College of Pharmacy, Moga, Punjab, India
2
Department of Pharmaceutical Chemistry, College of Pharmacy, National University of Science and Technology, Bousher, Muscat, Oman

Abstract

Pomegranate (Punica granatum) represents a major source of polyphenolic compounds. It has been gaining importance because of varied pharmacological activities attributed to its phytochemicals. This comprehensive review aimed to provide an exhaustive list of phytoconstituents found in pomegranate fruits, peel, flowers, and seeds along with their pharmacological activities. The potential of its different parts of pomegranate in treating diseases and the underlying mechanisms are discussed. The active phytochemical in the extract of pomegranate including flavonoids and tannins (ellagitannin and gallotannin) is responsible for the antiinflammatory, antioxidant, antiangiogenic, and antiaging properties. These have resulted in increased patent filing in the last decades by different research groups. Bioactive constituents of pomegranate have industrial application in conversion into value-added foods. Its reducing properties have wide applicability in the preparation of nanoformulations. A role of its phytoconstituents in environmental pollution management may prove to be safe alternative compared to using pesticides or other chemicals. An emphasis has been given on the several human clinical trials evaluating its pharmacological significance. Most of the study showed no evidence of the acute or subacute toxicity. Its bioactive phytoconstituents may be developed into drugs to prevent various diseases, but detailed clinical trials are still lacking to prove its safety and efficacy.

Keywords

Antioxidant activity; Clinical trials; Phytoconstituents; Polyphenols; Punica granatum L

11.1. Introduction

Punica granatum L. (family: Punicaceae or Lythraceae) is an ancient, long-living fruit-borne small deciduous tree. There are more than 1000 cultivars reported worldwide [1]. The cultivars are expanded from the Mediterranean region to Middle East and to American Southwest, California, and Mexico in the New World [2]. The common vernacular names of the plant in different languages are Anar (Hindi/Punjabi) [3], Dulim (Persia), Rumman (Arabic), Gangsalan (Indonesia), Granatapfel (German), Grenade (French), Melogranato (Italian), and Carthage (Romans) [4]. P. granatum belongs to closely related families of Myrtaceae, Lythraceae, Sonneratiaceae, Lecythidaceae, Rhizophoraceae, Combretaceae and Melastomataceae L. It is a deciduous shrub grown with a height up to 8m tall [5]. It is one of the oldest known edible fruits. The pomegranate fruits are considered as large berry, composed of mainly three parts: (i) peels forming interior network, around 40%–45%, (ii) the seeds which form around 3% of fruit weight, and (iii) oils (red color juice) forming about 30% of fruit weight surrounded around the seed in sac. The red color of the pomegranate juice is because of the anthocyanin present (3-mono- and 3,5-diglucosides of cyanidin, delphinidin, and pelargonidin) [6]. As per Viuda-Martos, the arils part comprises 40%, seeds 10%, and the remaining fruit is of 50%. The 50% of the fruit weight is because of the multiple seeds, embedded in white membranous, pericarp, thick skin [7]. The peels and seeds are agroindustrial waste having high contents of tannins and polyphenolic compounds. It is a source of glycosides, phytosterols, terpenoids, gallic acid, ellagic acid, pelargonidin, cyanidin, delphinidin, gallocatechin, etc. [8].

The tree can be further divided into various parts such as fruit, flower, leaves, bark, stem, and roots. History of pomegranates is reported back in 800-year-old documents in Judaism, Christianity, Islam, Buddhism, and Zoroastrianism [9]. Afghanistan, especially Kandahar is famous to produce a high-quality pomegranate [10]. Pomegranate, now a days, is cultivated all over the world with Iran as one of the largest producers [11]. The pomegranate plants can grow up to a height of 1600m from sea and in different climatic conditions. The major factor in the cultivation of pomegranate is its sensitivity to coldness. In cold conditions (less than 12°C), pomegranate fruits are damaged as for ripening it needs long hot summer conditions [12,13]. Mostly geographical regions with hot and long summers having sandy clay are best suited for high-quality pomegranate cultivation.

11.2. Morphology of Punica granatum

Fruit: Fruit is slightly yellowish to red in color. In some varieties, it is slight yellowish to green, and in rare cases, it is purple in color (Fig. 11.1). The weight of a pomegranate fruit varies from 200 to 800gm, and its diameter is around 5 to 20cm [13]. The fruit is made up of three parts that include peel, seed, and juice covered around the seeds in sac. It is the most consumed part of the plant. Leaves: The leaves are found as reciprocal in new sprouting branches while integrated in spores. Leaves contain unique tannins, glycosides of apigenin along with calcium and iron in high amount [14,15]. Seeds: Seeds are triangular in shape found trapped in aril part of fruit and are albumin free [10]. It generally contains 80% fatty acids with high amount of cis-9 and trans-11 fatty acids along with linoleic acid. The 99% of these fatty acids is triglycerol [16]. Flowers: Generally green to yellowish green in color found in four to five flowers in a group sometimes.

Figure 11.1
Different Parts of Pomegranate Fruit

11.3. Nutritional and medicinal importance

The pomegranate fruit is generally consumed directly in the form of seeds or in the form of fresh juice. Its juice is also used as a flavoring agent in jellies along with coloring agents [17,18]. Beside the nutritional value of its fruits, it also possesses a significant medicinal importance and can play a key role in health benefits for humankind [19]. Nowadays, a lot of research is oriented toward medicinal importance of pomegranate in human health (Fig. 11.2). The fruit is rich in phenolic compounds that possess potent antioxidant power [20,21]. In Ayurveda, pomegranate itself is considered as pharmacy within. The pomegranate juice is a good tonic for throat and heart. It is also observed to help in stopping gum bleeding and used in the treatment of hemorrhoids [22]. According to the oldest medical writing Eber's Papyrus (1500 BCE), the plant was used for the treatment of tapeworm and other parasitic infestation. The roots of pomegranate are also reported to possess anthelmintic properties [23]. The peels are found to possess significant astringent properties and are reported for the treatment of diarrhea [24]. Pomegranate juice also acts as tonic for blood [25]. P. granatum extracts of various parts of tree are also found to possess good antibacterial [26], antiinflammatory [27], anticancer [2], vasculoprotective effects [28], immunomodulatory properties [29], etc. The flavonoids and tannins reported in pomegranate fruits are reported to exhibit in vitro anticancer properties [30]. Luteolin and naringenin, two phytochemicals found in fruit part, are found to give same kind of effect as an enzyme secreted in women during pregnancy. It is also found to have effect on estrogenic receptors and can inhibit the estrogen-dependent breast cancer cell growth [31,32]. Ellagic acid found in peel part of the pomegranate is found to possess antiviral, antioxidant, and skin-bleaching properties. In Japan, it is used as an antioxidant in food products [33].

Figure 11.2
Health Benefits of Pomegranate

In addition to use in cosmetics [34,35], pomegranate is also reported to help in the treatment of AIDS [36], hormone replacement therapy [37], oral hygienic product [38], ophthalmic ointment [39], and as an adjuvant to increase the bioavailability of radioactive dyes [40]. Over the last few years, exhaustive research made use of its traditional properties to develop modern treatments such as the use of antioxidant properties in preventing inflammation and chemoprevention. The recent developments in chemistry and the extraction of phytoconstituents from pomegranate along with their pharmacological properties are discussed in this chapter.

11.4. Economic importance

P. granatum is produced on a large scale all over the world with approximately over two million tons per year, but around only 10% of its production enters the international trading, which is also further influenced by various factors such as domestic consumption to international space [41]. But, as compared to other fruit commodities, there is no reliable statistics available. Iran and India are the largest producer of pomegranate all over the world and supply fresh fruits as well as juice to European and other markets. There is high demand of fresh juice of pomegranate in market. Apart from it, flavored candies and wine are also in high demand. The demand of fresh fruit is also increasing day by day in international market due to well-established health benefits. Thus, there is a wide opportunity for cultivar countries to supply the fresh fruits to these markets during off-season depending on storage and quality as per region. The major chemical constituents include β-sitosterol and tannins and used in stomach disorders and diarrhea. The microscopic green color leaf powder showed the absence of trichome with prismatic or rosette crystals [3]. Other polyphenols include flavonoids, anthocyanins, hydroxy cinnamic acid, and hydroxybenzoic acids. It is also a rich source of fatty acids such as linoleic, palmitic, stearic, and arachidic acids.

11.5. Phytochemicals present in various parts of pomegranate

11.5.1. Leaf extract

The leaf ethanolic extract was characterized into three categories of metabolites namely phenolics, flavonoids, and triterpenes with the help of high-performance liquid chromatography coupled to photodiode array and mass spectrometry (HPLC-PDA-MS/MS) and gas chromatography–mass spectrometry (GC-MS) after derivatization with bis(trimethylsilyl)trifluoroacetamide [42,43]. Also, with the help of fast atom bombardment-mass spectrometry and nuclear magnetic resonance techniques, new unique phenolic compounds were identified. HPLC and spectroscopic analysis of the leaf n-hexane extract showed the presence of carotenoids, e.g., lutein and β-carotene as 3.652 and 1.915mg/g. The saponifiable and unsaponifiable fractions of n-hexane identified with the help of GC/MS are represented in Table 11.1 [44]. HPLC analysis of the P. granatum leaves’ hydroalcoholic [45] and ethyl acetate extract showed the presence of coumarins, flavonoids, and phenolic acids [46]. The compounds identified in the leaf extract are represented in Table 11.1.

Table 11.1

Some important compounds identified in the leaf and peel extract of Punica granatum.

S. No | Part used | Compound name | Technique used | Reference

1.

Leaf part (ethanolic)

Gallic acid, coumaric acid hexoside, brevifolin-carboxyl acid, ellagic acid, glucoside, galloyl-HHDPDHHDP-hexoside (granatin B), luteolin 7-oglucoside, ellagic acid, rutin, quercetin-3-O-glucoside, Apigenin-7-O-glucoside, Luteolin-4′-O-glucoside, luteolin, apigenin, oleanolic acid, betulinic acid, and ursolic acid

HPLC-PDA-MS/MS and GC-MS

[42]

2.

Leaf part (ethanolic)

N-(2′,5′-dihydroxyphenyl)pyridinium chloride, apigenin 4′-O-β-glucopyranoside, luteolin 4′-O-β-glucopyranoside, luteolin 3′-O-β-glucopyranoside, and luteolin 3′-O-β-xylopyranoside

FAB-mass spectrometry and NMR analysis

[43]

3.

Leaf (n-hexane extract)

Carotenoids, e.g., lutein (3.652mg/g), β-carotene (1.915mg/g), α-amyrin (1) acetate, ergosterol (2), and α-tocopherol (3)

HPLC and spectroscopical analysis

[44]

Phenanthrene, 1(Indanyl) adamantane, pregnan-3,20-dione, rhodoxanthin, cantaxanthin, 3,4-didehydro-1,2,7′,8′-tetrahydro-1-methoxy-2-oxocarotene, 1-methoxy1′-hydroxy-1,1′,2,2′-tetrahydro-3,4-didehydrolycopene), whereas unsaponifiable are hexadecanoic acid methyl ester, 11,14-octadecadienoic acid methyl ester, 9-octadecenoic acid methyl ester, octadecanoic acid methyl ester, cyclopropane-octanoic acid, 2-octylmethyl ester, heptadecanedioic acid, and 9 oxodimethyl ester

GC/MS

4. | Leaf (hydroalcoholic extract) | 3,3′-di-O-methylellagic acid, kaempferol (most abundant) (4), and kaempferol 3-O-glycoside | HPLC | [45]

5.

Leaf (ethyl acetate fraction)

Punicalin, ellagic acid derivate, galloyl-HHDP-glucose, castalagin derivative, castalagin derivative, granatin B, ellagic acid rhamnoside, kaempferol-3-O-glucoside, kaempferol derivative, and kaempferol-arabinoside

HPLC

[46]

6. | Peel extract (ethanolic) | Punicalagin, gallic acid, and ellagic acid | HPLC | [47]
Table Continued

S. No | Part used | Compound name | Technique used | Reference

7.

Ethanolic extract in microporous resin pretreated pomegranate husk

Hexahydroxydiphenoyl-valoneoyl-glucoside (HHDP-valoneyl-glucoside), galloyl-O-punicalin, rutin, hyperoside, quercimeritrin, kaempferol-7-O-rhahmano-glucoside, luteolin-3′-O-arabinoside, luteolin-3′-O-glucoside, and luteolin-4′-O-glucoside

HPLC-QTOF-MS

[48]

8. | Ethanolic fruit peel extract | 1,2,3,4-butanetetrol, dimethyl sulfone, 9-octadecenamide, and pentadecanoic acid | FTIR and GC-MS chromatogram | [49]
9. | Dried peel of the P. granatum was extracted with 80% methanolic solvent and fractioned in acetone | Novel chalcone derivatives (2E)-3-(4-hydroxy-3-pentylphenyl)-1-(2-hydroxy-4,5-di((E)-prop-1-enyl)phenyl)prop-2-en-1-one | UV–Visible, FT-IR, 1H NMR and Mass spectra | [50]

10.

Methanolic extract and fractioned with ethyl acetate

New pyrrolizine alkaloid namely punicagranine (5), novel alkaloid named as 5-(furan-3-carbonyl)-2,3-dihydro-1H-pyrrolizine-7-carboxylic acid

1D- and 2D-NMR and HR-ESI-MS spectra and then confirmed by X-ray diffraction

[51]

11. | Hydroethanolic extract peel extract | Polysaccharide galactomannan (PSP001) | Gel filtration chromatography with Sephadex G-200 | [52]
12. | Rind ethanol extract | Pyrrolidine-like alkaloid punigratane (6) | IR, MS, and NMR spectroscopic techniques | [53]
13. | Methanol-chloroform extract | Gallic acid (7), caffeic acid, vanillic acid, ferulic acid, quercetin (10), quinic acid (9), and kaempferol | HPLC and GC-MS | [54]

14.

Acetone extract of dried pomegranate peel

Punicalagin (11) and ellagic acid (8), brevifolin carboxyfolin acid, gallic acid, ferulic acid-hex, ellagic acid-hex, ellagic acid-deoxyhex, ellagic acid, punicalagin, gally-hexoside, catechin, HDDP-hex, punicalin, valoreic acid bilactore, ellagic acid dihex, caffeic acid hex

LC-ESI-QTOF-MS/MS

[55]

15. | Dried peel extracted with 70% acetone | Four hydrolysable tannins namely punicalagin, punicalin (12), strictinin (13), and granatin | Column chromatography combined with in vitro antiinflammatory-guided fractionation | [27]

11.5.2. Peel extract

The major bioactive compounds isolated from the ethanolic peel extract belong to ellagitannin and gallotannin. The phytocompounds were punicalagin, gallic acid, and ellagic acid [47]. The HPLC quantification of the 50% pomegranate peel extract showed the presence of punicalagin, punicalin, gallic acid, and ellagic acid as 56.78±0.45, 200.02±0.79, 5.52±0.03, and 20.93±0.09 (mg/g of extract) [56]. The ethanolic extract in microporous resin pretreated pomegranate husk by HPLC-QTOF-MS characterized a total of 50 polyphenols with 35 hydrolysable tannins and 15 flavonoids. Among them, nine compounds reported for the first time shown in Table 11.1 [48]. Mayasankaravalli et al. extracted 23 compounds from the ethanolic fruit peel extract. The major compounds were 1,2,3,4-butanetetrol, dimethyl sulfone, 9-octadecenamide, and pentadecanoic acid [49]. Recently, Rivas et al., used a supercritical fluid extraction technique using CO_2 to extract the active phytoconstituents from the pomegranate peel in good amount. The optimization of the extraction was done using Box–Behnken design combined with response surface methodology and the optimum temperature and pressure ought to be 46.5°C, (291bar) run for time 2.5h. The method considered to be valuable in achieving high yield of the compounds from pomegranate by-products [57]. A novel chalcone derivative (2E)-3-(4-hydroxy-3-pentylphenyl)-1-(2-hydroxy-4,5-di((E)-prop-1-enyl)phenyl)prop-2-en-1-one was isolated from the peel by Sudheesh et al. [50]. The compound was isolated after the dried peel of the P. granatum was extracted with 80% methanolic solvent and fractioned in acetone. Sun et al., identified new pyrrolizine alkaloid namely punicagranine from the peel of P. granatum. The novel alkaloid named as 5-(furan-3-carbonyl)-2,3-dihydro-1H-pyrrolizine-7-carboxylic acid was extracted from the methanolic extract and fractioned with ethyl acetate [51]. The five compounds quantified with the help of HPLC and GC-MS were gallic acid, caffeic acid, vanillic acid, ferulic acid, and quercetin from the P. granatum rind. The measured concentration among them is high for quercetin, i.e., 90.24941μg/mL [54]. The total tannin aqueous acetone fraction of P. granatum peel showed punicalagin and ellagic acid were present in abundant form. Other compounds identified in the total tannin contents of P. granatum with the help of LC-ESI-QTOF-MS/MS are represented in tabular form [55]. The compounds reported in peel extract of the pomegranate are represented in Table 11.1.

11.5.3. Pomegranate juice

The juice mainly consists of water (85%), sugar (10.6%), and small amounts of pectin, polyphenols, fatty acids, sterols, and minerals [58]. The pomegranate juice showed the presence of phenolics, and flavonoids compounds namely luteolin, ellagitannin, and epigallocatechin gallate (EGCG) [59]. The flavonoid and phenolic contents were found in rich quantities and calculated as 14.94±1.6mg GAE/g. The HPLC profile of the pomegranate fruit extract revealed the presence of catechin, caffeic acid, epicatechin gallate, cinnamic acid, benzoic acid, esculetin, hesperidin, and quercetin [60]. The ethanolic fruit extract of P. granatum showed the presence of ellagic acid, gallic acid, and punicalagin A and B [61]. It also contains major organic acids such as citric and malic acid [62]. The juice of pomegranate analyzed through LC-MS/MS showed the presence of chinic acid, malic acid, 3,30-di-O-methyl ellagic acid, ellagic acid hexoside, ellagic acid, tryptophan, and catechin [63]. Lee et al. isolated four hydrolysable tannins namely punicalagin, punicalin, strictinin, and granatin from the peel of P. granatum [64]. The HPLC results of the pomegranate juice showed the major polyphenol to be punicalin (0.6%), gallagic acid (2.6%), punicalagin (2.7%), gallic acid (5.3%), and ellagic acid (88.8%) [65].

11.5.4. Seed residue

Pomegranate seeds are rich sources of phospholipids such as lecithin, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol [66]. The pomegranate also has a high content of calcium (0.01%), magnesium (0.012%), potassium (0.236%), and phosphorous (0.036%) [67]. A total of 47 phenolic compounds were identified from the seed following juice extraction. The phytoconstituents present were characterized by HPLC-DAD-ESI-MSn as phenolic acid, hydrolysable tannins, monomeric flavonoids, anthocyanin, and proanthocyanin [68]. Very similar compounds were also isolated earlier by Fischer et al. [69]. A total of 48 compounds were reported among which nine are anthocyanin, two gallotannin, 22 ellagitannins, two galloyl esters, p-hydroxybenzoic acid, seven hydroxycinnamic acids, and one dihydroflavonol. The phytochemicals extracted from the pomegranate juice and seeds are shown in Table 11.2.

The LC-MS/MS analysis of the seeds extract showed the presence of 15,16-dihydroxy-9Z,12Z-octadecadienoic acid, methyl 2-[cyclohex-2-en-1-yl(hydroxy)methyl]-3-hydroxy-4-(2-hydroxyethyl)-3-methyl-5-oxoprolinate, p-hydroxybenzoic acid, and oxooctadecanoic acid (stearic acid) [63]. Other chemical constituents isolated from the seeds portion include ursolic acid, γ-tocopherol, sterols (stigmasterol, β-sitosterol, and campesterol), punicic acid (most abundant fatty acid in pomegranate seed total of 60% of the seed oil), and hydroxybenzoic acid (gallic and ellagic) [2]. He and coworkers extracted phenolic compounds with the help of subcritical water extraction from the pomegranate seed residues. The highest total phenolic contents were extracted at 220°C. The antioxidant activity by HPLC-ABTS
.+ radical system showed nine of the compounds to possess antioxidant activity. The pomegranate seeds contain a high percentage of lignin and in that phenolic acids are conjugated with polysaccharides, proteins by ester, and ether bonds. These get hydrolyzed with effect from the subcritical water at high temperature, and free phenolics are released from matrix. The 5-hydroxymethyl furfural (5-HMF) possesses antioxidant activity and generated as intermediates via Amadori rearrangement of Maillard reaction. The 5-HMF dramatically increased from 7.32 (μg/g DW) at 100°C to 110.14 (μg/g DW) at 220°C [70]. The presence of biologically active O-prenylated umbelliferone derivatives, e.g., auraptene and umbelliprenin detected with the help of UHPLC with UV/Vis detection by Fiorito et al. [71]. The ethanol seed extract yielded the highest concentration of auraptene (23) and umbelliprenin (24) as 1.99μg/g and 6.53μg/g. The metabolite umbelliferone (25) was also detected in the concentration of 0.67μg/g. The method was found to be efficient for the detection and quantification of oxyprenylated coumarins present in pomegranate seeds. The structure of some selected phytoconstituents is represented in Fig. 11.3.

Table 11.2

The list of phytoconstituents isolated from the seeds and juice of P. granatum.

Class | Phytoconstituents

Hydroxybenzoic acid and hydroxycinnamic acid

Protocatechuic acid (∗) (∗∗), trans p-Coumaric acid (∗) (∗∗), vanillic acid (∗), gallic acid (∗) (∗∗), brevifolin carboxylic acid (19) (∗), p-Hydroxybenzoic hexoside (∗), gallic acid pentoside (∗), vanillic acid-hexoside (∗) (∗∗), cis-Caffeic acid hexoside (∗) (∗∗), trans-Caffeic acid hexoside (∗) (∗∗), 5-O-Caffeoylquinic acid (∗) (∗∗), ferulic acid hexoside (∗) (∗∗), caffeic acid hexoside derivative (∗) (∗∗)

Flavanoids

(+)-catechin (∗), dihydroxygallocatechin (∗), naringenin hexoside (∗), quercetin 3-O-rhamnoside (∗), quercetin hexoside (∗), kaempferol 3-O-glucoside (∗), cis-Dihydrokaempferol hexoside (∗) (∗∗), trans-Dihydrokaempferol hexoside (∗)

Tannins

Ellagic acid (∗) (∗∗), ellagic acid-derivative I (∗) (∗∗), ellagic acid-derivative II (∗) (∗∗), ellagic acid pentoside (∗) (∗∗), ellagic acid deoxyhexose (∗) (∗∗), ellagic acid hexoside (∗) (∗∗), valoneic acid bilactone (18) (∗), valoneic acid bilactone II (∗), HHDP hexoside I (∗) (∗∗), HHDP hexoside II (∗), digalloyl hexoside (∗) (∗∗), procyanidin dimer (∗), HHDP diglucoside (∗), galloyl-HHDP-hexoside (corilagin) (∗) (∗∗), gallotannin (∗), ellagitannin (∗∗), bis-HHDP-hexoside (pedunculagin I) (∗∗) (15), digalloyl-HHDP-hex (pedunculagin II) (∗∗), galloyl-bis-HHDP-hex (casuarinin) (∗∗), flavogalloyl-HHDP-gluconic acid (lagerstannin B) (∗∗), lagerstannin B derivative (∗∗), castalagin derivative (∗∗), galloyl-HHDP-DHHDP-hex (granatin B) (∗∗), brevifolin carboxylic acid (∗∗), gallagyl ester (∗∗), gallagyl hexoside (punicalin) (∗∗), HHDP-gallagyl-hexoside (punicalagin) (∗∗), galloyl-HHDP-gluc (lagerstannin C) (∗∗) (17), digalloyl-HHDP-gluc (punigluconin) (16) (∗∗)

Anthocyanins

Cyanidin-3-O-pentoside (∗), pelargonidin-3-O-glucoside (∗), cyanidin-3-O-glucoside (∗), delphinidin-3-O-glucoside (∗), cyanidin-pentoside-hexoside (∗), cyanidin-3-O-rutinoside (∗), cyanidin 3,5-O-diglucoside (∗), delphinidin 3,5-diglucoside (∗), (epi) Afzelchin-delphinidin-3-O-hexoside (∗), (epi) gallocatechin-pelargonidin 3-ohexoside (∗), (epi) Afzelchin-delphinidin-3-O-hexoside (∗), (epi) gallocatechin-cyanidin-3-O-hexoside (∗)

The compounds represented in bracketed (∗) were reported by Ambigaipalan et al. [68] and (∗∗) by Fischer et al. [69].

11.5.5. Pomegranate flower

Pomegranate flowers have been reported to be a source of triterpenoids including ursolic acid [(3β)-3-hydroxyurs-12-en-28-oic acid] (20) and its isomers and oleanolic acid [(3β)-3-hydroxyolean-12-en-28-oic acid] (21), whereas only ursolic acid has been isolated from pomegranate seeds. The gallic acid (3,4,5-trihydroxybenzoic acid) (22) reported to be present in pomegranate peel, juice, and flower in free form, glycosides, ellagic acid, and ellagitannin [72]. The flower part also contains glycated anthocyanins, e.g., pelargonidin 3,5-diglucoside and pelargonidin-3-glucoside while leaves, roots, and stem contain apigenin, punicalin, punicalagin, and luteolin [73]. The methanolic extract of pomegranate also reported to contain flavones such as luteolin and tricetin [62]. The presence of asiatic and maslinic acid is also reported [2]. The pomegranate flower extract by HPLC-QTOF-MS/MS showed the presence of galloyl-HHDP-glucoside, granatin A, digalloyl-glucoside, tri-O-galloyll-glucoside, galloyl-HHDP-glucoside, granatin B, ellagic acid, ellagic acid derivative, ellagic acid rhamnoside, quercetin, quercetin-O-glucoside, kaempferol, and luteolin [74].

11.5.6. Bark extract

The major alkaloids present in the stem bark and root are pelletierine, pseudopelletierine, and N-methyl pelletierine [75]. Other alkaloids hygrine and norhygrine are also reported from the bark of the root of P. granatum [76]. Marsoul et al. reported that the polyphenolic contents of the P. granatum barks were 272.82±3.05mg/mg gallic acid equivalent; GAE/g sample and the minor was total flavonoids 38.26±1.78mg/mg quercetin equivalent; QE/g sample [77]. The metabolomic profiling of the originally grown pomegranate samples showed that its juice contains high amount of acetic acid, alanine, arginine, fumaric acid, gamma-aminobutyric acid, galactose, glutamine, histidine, isoleucine, lactic acid, leucine, malic acid, mannose, methionine, phenylalanine, proline, sucrose, threonine, trigonelline, tyrosine, and valine, whereas conventionally grown pomegranate showed the presence of high amount of acetone, aspartic acid, and ethanol. The presence of phytoconstituents in pomegranate grown in organic conditions has more nutritional quality than pomegranate grown conventionally [78]. The seeds portion includes ursolic acid, γ-tocopherol, sterols (stigmasterol, β-sitosterol and campesterol), punicic acid (most abundant fatty acid in pomegranate seed total of 60% of the seed oil), and hydroxybenzoic acid (gallic and ellagic) [2].

Figure 11.3
Chemical Structures of Some of the Selected Phytoconstituents Found in Pomegranate

11.6. Pharmacological activities of pomegranate

Pomegranate is one of the most important ornamental crops. Its medicinal properties have been known to humankind since ancient time. The biological and pharmacological uses of this plant are attributed to the presence of diverse classes of secondary metabolites. Some of its important pharmacological activities explored in the past decade are discussed in the section.

11.6.1. Anticancer activity

The anticancer potential of pomegranate is due to its rich polyphenolic content. The plant extract has been shown to act as antiproliferative, antimetastatic, and antiinvasive against different cancer cell lines in vitro, in vivo, and in human interventional (clinical) trials [79]. The angiogenic inhibitory activity of pomegranate is linked to the presence of polysaccharide galactomannan that reduced the neovascularization in chick embryos. The peritoneal angiogenesis assay in BALB/c mice further supported its angiogenic inhibition. It also inhibited the invasion, migration, and clonogenic capacity in human murine cancer cells. The polysaccharide downregulated the VEGF, MMP-2, and MMP-9 and lead to an elevation in TIMP-1 and TIMP-2 levels. Along with vincristine, it showed better survival rate and decreased metastatic index in C57BL/6 mice model [52]. The pomegranate fruit extract also exhibited the antiangiogenic effect against human pancreatic (Suit-2) and colon (colo205) cancer cell lines developed in the chick chorioallantoic membrane (CAM). The extract was found to reduce the tumor weight and hemoglobin contents in the studied model. It also reduced the angiogenic effect of fibroblast growth factor (FGF2) [80]. Rahimi et al. studied the effect of ellagic acid present in pomegranate against inflammation and cancer. Human and preclinical studies indicated it to have immunomodulatory action produced due to its ability to suppress humoral immunity and regulate T-cell function [81].

The potential of pomegranate in the treatment of breast cancer has been extensively studied. The breast cancer treatment relies on the inhibition of the mechanism that directs the estrogen activity. It also includes the antagonism of estrogen synthesis or estrogen receptors. Pomegranate polyphenols showed strong anticancer activities, induces cell cycle arrest in Go/G_1 phase, inhibited invasion, migration, and viability of breast cancer cells. Bishayee et al. studied the chemopreventive effect of the pomegranate emulsion against 7,12-dimethylbenz(a)anthracene (DMBA) induced rat mammary carcinogenesis. The animals were orally administered with pomegranate extract (0.2–5.0g) starting with two weeks before treatment and continue up to 16 weeks after the DMBA treatment. The pomegranate extract led to a reduction in mammary tumor incidence, decreased tumor burden, suppressed cell proliferation, and promoted apoptosis. It also confirmed increased Bax and decreased Bcl2 in immunohistochemical studies. The extract led to upregulate Bad, caspase-3, caspase-9, cytochrome complex (cyt c), and poly (ADP ribose) polymerase. The polyphenols-rich fractions of the pomegranate were evaluated for in vitro for chemopreventive and adjuvant therapeutic potential in human breast cancer [82]. The polyphenols from the fermented juice, pericarp, and whole seeds oil inhibited the aromatase activity by 60%–80% and 17-β-hydroxysteroid dehydrogenase type 1 from 34% to 79% in decreasing order of fermented seed oil, fermented juice polyphenols, and pericarp polyphenols (conc 100 to 1000μg/mL). In both the cell lines of MCF7 and MB-MDA-231 cells, fermented pomegranate polyphenols showed twice the antiproliferative effect as compared to the fresh pomegranate juice polyphenols. Pomegranate seed oils showed 90% inhibition (100μg/mL) in the proliferation, 75% invasion of MCF7 (10μg/mL), and 54% apoptosis in MDA-MB-435 estrogen-negative breast cancer cell lines (50μg/mL) [83]. Pomegranate extract was evaluated for experimental menopausal syndrome in ovariectomized mice. After two weeks administration of pomegranate seeds and juice extracts, it was found to prevent the loss of uterus weight, normalized the decreased bone mineral density, and shortened the immobility time (antidepressant). Further, it increased the bone volume and trabecular number in the pomegranate-treated groups [84]. Ellagic acid present in pomegranate could result in potent antiestrogen effect in MCF-7 breast cancer cell lines like pure estrogen antagonist ICI182780, IGFBP-3 levels. It also induces nodule mineralization. At low concentration, ellagic acid (10
−7 to 10
−9
M) showed significant estrogenic activity through Erα, whereas it showed complete antagonism by ERβ. Ellagic acid increased the alkaline phosphatase activity and stimulated the mineralization at a concentration of 10
−8 to 10
−9
M acting as a natural selective estrogen receptor modulator (SERM) [85]. Another mixed effect, i.e., both estrogenic and antiestrogenic actions of ellagic has been reported. The ellagic acid treatment increased hTERT α+β+mRNA, whereas it decreased the 17β-estradiol-induced increase in hTERT α+β+mRNA. The ellagic acid downregulated 17β-estradiol-induced hTERT α+β+mRNA expression and therefore might exert the chemopreventive effects in breast cancer [86]. In another study, it was observed that genistein treatment led to cell cycle arrest and improvement in mitochondrial functionality in T47D cell with low (Erα/ERβ) but not in MCF-7 (high ERα/ERβ) and MDA-MB-231 (ER-negative one) [87].

11.6.2. Neuroprotective activity

Fatima et al. studied the neuroprotective effect of the P. granatum against scopolamine-induced cognitive deficit in rats. The results showed that the neuroprotective effect of pomegranate juice extract (500mg/kg orally) was better than rosuvastatin (10mg/kg) and was comparable to the standard drug donepezil (0.5mg/kg orally) in Wistar rats. Thus, the pomegranate juice extract has a protective role in learning, memory, and behavior in scopolamine-induced amnesia model of Alzheimer's disease [88]. Kasi et al. explored different available fruit juices for inhibition of the formation of amyloid fibrils. Among them, pomegranate juice showed the most effective inhibition of amyloid aggregation in turbidity measurement and Congo red binding assays. The pomegranate juice reduced the phenylmethanesulfonyl-trypsin amyloid-like fibrils formation by 3.7% as compared to without sample [89]. Amri et al. demonstrated the neuroprotective effects of the pomegranate extract mediated via inhibition of the cholinesterase enzyme and activating antioxidant potential. The study was done on high-fat, high-fructose–induced obesity in rats. Among the different extract of the pomegranate (seed, leaves, and juice), the leaves extract showed the highest cholinesterase inhibitory activity with IC_50 of 151.85mg/mL. It decreased the cholinesterase percentage activity in brain from 17.4% to 6.4%. The extract controlled lipid profile accumulation in brain and body. It decreased the levels of brain oxidant markers such as malondialdehyde (MDA), protein carbonylation, and increased superoxide dismutase (SOD) and glutathione peroxidase levels [90]. The peel extract of the P. granatum was evaluated for inflammation, oxidative stress, biomarkers of neuroplasticity, and spatial memory in degeneration model of the mice. The peel extract showed the presence of high phenolic contents (21.25±0.16mg gallic acid/g) and flavonoids (7.60±0.12mg quercetin/g). The concentration of the punicalagin was found to be in concentration of 13.74mg/g of extract. The male C57Bl/6 mice group was treated with P. granatum extract (800mg/kg/day) in the mice infused with amyloid-β peptide (35 days with amyloid-β peptide, Aβ). The pomegranate extract-treated groups showed a reduction in the activity of acetylcholinesterase enzyme and amyloid plaque density and increase in the expression of brain-derived neurotropic factor (BDNF). The increase in neurotropin density was prominent in cortex (19.60±4.87pg/μg) and hippocampus (25.21±3.98pg/μg) as compared to the control. The neuronal activity and BDNF density are important for the survival and proper functioning of the dopaminergic and cholinergic neurons present in the neurons of the hippocampus and cortex. Further reduction in the concentration of proinflammatory cytokines (tumor necrosis factor α [TNF-α]) and lipid peroxidation was also noted. A slight increase in the quantity ofMDA and nonsignificant increase in the SOD activity were observed in the pomegranate extract-treated groups [91]. Studies showed that ellagic acid (dimer of gallic acid) decreases the advancement of neurodegeneration. The mechanism was found to be its antioxidant effect, iron chelating, and protecting effect on mitochondria [92]. The reactive microglia with extracellular plaques of Aβ is indication of the Alzheimer's disease. The memory was evaluated by male amyloid precursor protein/presenilin 1 (APP/PS1) in the male transgenic mice (12-month-old). The groups were fed with free access of the normal chow, drinking water, and pomegranate extract (6.25mL/L) for three months. It decreased the length of the time to escape in Barnes maze and showed lower TNF-α and lower nuclear factor of activated T-cell transcriptional activity. It also decreased the Aβ plaque deposition and microgliosis. In the cell culture, the compound ellagic acid and punicalagin decreased the Aβ-stimulated TNF-α secretion in murine microglia [93]. Braidy et al. studied the supplementation of 4% pomegranate extract for 15 months to standard chow diet on neuroinflammation and synaptic plasticity in mice (APPsw/Tg2576) brain. The HPLC analysis confirmed the presence of the ellagitannin such as punicalagin and punicalin and flavonoids quercetin and kaempferol derivatives in the pomegranate extract. The results explained that it inhibited the neuroinflammation and activated phosphoinositide-3-kinase-Akt-mammalian target of rapamycin (mTOR) signaling pathway. It also led to the phosphorylation of the calcium/calmodulin-dependent protein kinase IIa (improved pCaMKIIα/CaMKIIα), enhanced the autophagy and cyclic AMP-response element binding protein (improved pCREB/CREB), and thus attenuated the deficits in memory and cognition. Further, it reduced the loss of synaptic structure protein (PSD-95, Munc18-1, and SNAP25) (maintained the normal plasticity) and β-site cleavage of APP. It also significantly reduced the BACE1 expression as inhibition reduced the Aβ levels in Alzheimer's disease. Another activation is a PI3K-Akt-mTOR pathway with the pomegranate extract which was reported to reduce because of the increase in APP processing and Aβ peptides [94]. Their group also showed the beneficial effect of pomegranate juice combined with fig and dates on suppressing the inflammatory cytokines and delay in the formation of senile plaques in APPsw/Tg2576 mice. The combination also helped in decreasing the brain Aβ1-40 and Aβ1-42 contents. Thus, it showed the beneficial effects of pomegranate in reducing inflammation in aging and neurodegenerative-related diseases [95].

11.6.3. Antioxidant activity

Different solvent extracts viz., hexane, dichloromethane, ethyl acetate, ethanol, and methanol of P. granatum leaves were evaluated for antiinflammatory, antioxidant, and cytotoxic potential. The methanolic extract showed the most potent antioxidant activity with IC_50 value of 5.62mg/L (2,2-diphenyl-1-picrylhydrazyl [DPPH]), 1.31mg/L (ABTS assay), and antiproliferative activity against MCF cancer cell lines (IC_50
=31mg/L). The ethanolic extract showed the maximum acetylcholinesterase (IC_50
=6.20mg/L), lipoxygenase (LOX) (IC_50
=14.83mg/L), and butyrylcholinesterase (IC_50
=2.65mg/L) inhibition [96]. The hydrolyzed contents, mostly the free phenolics from the pomegranate juice and seeds, were evaluated for antioxidant activity. The phenolics phytoconstituents were able to scavenge the hydroxy and DPPH radicals. The seed's total phenolics and scavenging activity were greater than juice fraction. These phenolics also demonstrated α-glucosidase and lipase inhibitory activity and thus seems to be helpful for diabetic and obese patients [68]. The polysaccharide galactomannan (PSP001) obtained from the seed oils and fruit rind of P. granatum has potential of antioxidant, immunomodulatory, and anticancer effects [73]. The green solvent extraction of protein, bioactive peptides, and phenolic compounds from pomegranate using pressurized liquids (pressurized liquid extraction [PLE]) or deep eutectic solvents showed different composition and spectrum of activity. The pressurized liquids exhibited high antioxidant and hypocholesterolemic extracts, whereas deep eutectic solvents protein extracted hydrolysates with high antihypertensive capacity. Peptide amounts were higher in the hydrolysates of deep eutectic solvents, whereas phenolic compounds were higher in PLE extracts. Only one peptide (HPVLV) was found in hydrolysate of alcalase, while four (VTYDYYEL, LSGGPMVVAHE, MPVVAEH, and ARAR) in thermolysin. The phenolic constituents were mainly punicalin and punicalagin. Other phytoconstituents in small amount were galloyl-HHDP hexoside, ellagic acid-hexoside, and ellagic acid [97]. Findings of another study confirmed antioxidant, antidiabetic, antihypertensive, and antiobesity effects of ethanolic extract of P. granatum fruits peel. The extract showed the highest inhibition of α-glucosidase, lipase, angiotensin converting enzyme, and potent 1, 1-diphenyl-2, 2, picrylhydrazyl scavenging [49].

The presence of polyunsaturated fatty acid called as punicic acid (omega-5; w-5) present in significant amount in pomegranate seed oils works as a potent antioxidant. It showed beneficial effect against pathological liver conditions tested in mice C57BL/6N fed with high-fat diet in hepatic steatosis. The punicic acid increased the fatty acid oxidation in primary hepatocytes and showed antioxidant gene expression [98]. Ammar et al. studied the pomegranate juice effect on oxidative markers enzymes after weightlifting exercise. It was noted that pomegranate juice attenuated the increase in MDA and increased enzymatic action of catalase and glutathione peroxidase. There has also been decrease in nonenzymatic uric acid and total bilirubin antioxidant response just after three minutes after the exercise. There was seen recovery kinetics in MDA and other antioxidant enzymes after 48h [99]. The oxidative stress on lungs was induced by the cigarette smoke (1–3-month five days exposure in a week) in male C57BL/6J mice, and the effect of the pomegranate juice was evaluated. Pomegranate juice was given daily at a dose of 80μmol/kg, and the inflammatory markers, apoptosis, and oxidative stress were studied. In vitro test cell viability assay was carried out in A549 cells pretreated with pomegranate juice for 48h. The pomegranate juice reduced the expression of the inflammatory markers, apoptosis, and oxidative stress. The juice of pomegranate further reduced the TNF-α and normalized lungs architecture. Human alveolar cell culture exposed with cigarette smoke showed a less damaging effect when supplemented with pomegranate juice [100]. Another study by Hosari et al. also showed that pomegranate juice prevents the formation of lungs nodule to chronic cigarette smoke exposure in mice. Here the animals were exposed for total five months (five days per week). The histological analysis, mitotic activity (measuring the PHH3 antibodies), and hypoxia-inducible factor-1α (HIF-1α) expression was studied in the cigarette smoke and pomegranate- exposed group. Pomegranate juice significantly reduced the nodule formation and attenuated the expression of PHH3 antibodies and HIF-1α [101].

11.6.4. Lipid-lowering activity

The hydroethanolic pomegranate peel extract was investigated for their lipid-lowering effect against high lipid-fed male rats by Sadeghipour et al. The pomegranate extract was given intraperitoneally in concentration of 50, 100, 200, and 300mg/kg body weight for 23 days, and the levels of serum cholesterol, triglycerides, low-density lipoprotein (LDL), high-density lipoprotein (HDL), aspartate transaminase (AST), and alanine transaminase (ALT) were evaluated. The treatment decreased the weight of rats and significantly decreased total cholesterol 81±7 (control 110±11), triglycerides 210±27 (control 475±11), LDL-C 61±7 (92±6), alkaline phosphatase 1049±78 (983±21), AST 1233±36 (1538±47), and ALT 1290±54 (1553±32) mg/kg as compared to the saline control. It increased the HDL-C level and decreased the liver damage in high lipid diet-fed rats in histopathological studies [102].

11.6.5. Antibacterial activity

The n-hexane extract of P. granatum showed strong antimicrobial activity against nine microbial pathogens (Pseudomonas aeruginosa, Escherichia coli 100μL, Salmonella spp, Staphylococcus aureus, Listeria monocytogenes, Ent. Fecalis, Bacillus cereus, Candida albicans, Aspergillus niger). All the microbial pathogens were inhibited by 150μL of the extract at 60min. Among the tested microbial strains, the antibacterial effect against S. aureus was found to be most potent (MIC value of 50μL after 60min). The obtained extract showed no evidence of toxicity [44]. Another study provided an evidence of moderate activity of hydroalcoholic extract of P. granatum against planktonic and biofilm cells of S. aureus including both methicillin-resistant and methicillin-sensitive strain of S. aureus. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were 125–250μg/mL and 500–1000μg/mL, respectively [8]. The presence of new pyrrolidine namely punigratane was evaluated for MDR Klebsiella strain and found to show high efflux pump activity [53]. Gosset-Erard et al. extracted water, water:ethanol (1:1), ethanol, acetone, and heptane fraction from the peel of pomegranate var. Zheri and Hicaznar. The extract of water and water:ethanol showed significant antimicrobial activity. The active constituents from the ethanolic extract were identified by the mass spectrometry to be punicalagin. The α/β form of the punicalagin was found to exist in the ratio of 3:7, and both forms were active with MIC value between 0.3 and 1.2μg/mL [103]. The novel derivatives of chalcone (2E)-3-(4-hydroxy-3-pentylphenyl)-1-(2-hydroxy-4,5-di((E)-prop-1-enyl)phenyl)prop-2-en-1-one showed prominent activity against LOX and cyclooxygenase (COX) and may act as potential antiinflammatory agents [50]. The compound showed enzyme activity LOX as 78.48 as compared to standard vanillin 89.87. The maximum COX activity was 25μM/min as compared to aspirin (38μmol/min) and nimesulide (29μmol/min).

11.6.6. Antiviral activity

The ethanolic leaf extract and the isolated fractions compound, ellagic acid are active against Zika virus infection. Ellagic acid showed antiviral activity with an EC_50 value of 30.86 and 46.23μM against MR766 and HPF2013 Zika virus strain. The mechanism of action displayed Zika virus progeny reduction [42]. The ethanolic peel extract showed antiviral activity with significant inhibition against herpes simplex virus (HSV)-2 at a dose of 62.5μg/mL. The compound punicalagin showed 100% anti-HSV-2 activity at a dose of 31.25μg/mL [47]. An aqueous extract of the leaves of P. granatum showed potential antiviral activity against HHV-3 and was comparable to the standard drug acyclovir. The leaf phytochemicals interacted with HHV-3 protease and may interfere with the capsid assembly of HHV-3. Docking studies confirmed the better binding interaction of phytochemicals granatin A, strictinin, and corilagin, but the binding energies were found to be lower than apigenin, isoquercetin, and luteolin [104]. The pomegranate rind extract along with different zinc preparations (zinc sulfate, zinc citrate, zinc stearate, and zinc gluconate) showed anti-Herpes simplex virus (HSV-1) by fourfold. However, punicalagin alone showed eightfold greater virucidal activity (5.93±0.35) as compared to pomegranate extract (0.76±0.66). The pomegranate extract (EC_50
=0.556±0.39μg/mL) showed 1000-time more potent antiviral activity mass to mass basis against HSV-1 as compared to acyclovir (EC_50
=0.182μg/mL). Further, it also showed greater antiviral response against acyclovir-resistant strain of HSV (EC_50
=0.016±0.009μg/mL), whereas punicalagin and ellagic acid showed no antiviral activity at tested concentration (IC_50
>100μM) [105].

11.6.7. Antileishmanial and anticoccidial activity

The antileishmanial activity of the pomegranate juice was evaluated by Alkathiri et al. It was found that antileishmanial activity of EGCG was greater than the standard drug miltefosine. Ellagitannin works by macrophages activation, induces NO and tumor necrosis factor-gamma, and with the help of cytokines kills the parasites and provides strong host defense system. Luteolin inhibited the extracellular promastigotes. The P. granatum juice reduced the size of cutaneous leishmaniasis lesions as compared to the untreated mice, and it was because of the enhancement of the antioxidant enzymes [59]. The crude methanolic extract of fruit peel of P. granatum showed maximum anticoccidial activity as compared to the standard amprolium. The anticoccidial activity was because of the alcohol-soluble ingredients present. It was found there were weight gain parameters and better feed conversion ratio and reduction of oocyst output in crude methanolic extract-treated chicken. This was related with the decrease in the intestinal inflammation and injuries [5].

11.6.8. Antiinflammatory activity

The phytoconstituents present in the ethanolic extract (ellagic acid, gallic acid, and punicalagin A and B) were found to inhibit the levels of inflammatory mediators, i.e., lipopolysaccharide (LPS)-induced NO, PGE-2, and IL-6. The nitrite production by the phytoconstituents was decreased in a dose-dependent manner, and highest inhibition was observed in case of ellagic acid. Similarly, LPS-induced PGE-2 and IL-6 production were suppressed in a dose-dependent manner. Thus, these phytoconstituents were suggested to be responsible for the antiinflammatory potential of P. granatum [61]. Punicalagin (25, 50, and 75mg/kg body weight) was evaluated against collagenase-induced intracerebral hemorrhage (ICH) induced inflammation, and oxidative stress in male Sprague–Dawley (SD) rats. The extract reduced the cell damage and inflammatory cell infiltration. It showed improvement in the brain architecture and BBB integrity and higher protective effects at a dose of 50 and 75mg/kg. The antioxidant status of the punicalagin showed enhanced effect through the activation of the Nrf-2/ARE/HO-1 signaling pathway. The increase in Nrf2 translocation was enhanced from 139% (ICH control) to 181.98% at 75mg punicalagin dose. It decreased the ROS levels to 115.09% from 405.58% (ICH control) at a dose of 75mg/kg and MDA levels from 18.05 (ICH control) to 1.1nM/mg of protein. The levels of the SOD and CAT activities were also increased in the punicalagin-treated groups. It also increased the HO-1 expression from 127% (control group) to 174%. Further, it also increased the activity of NF-κB levels and inversed the upregulation in IL-6, IL-18, and IL-1β. The punicalagin reduced the expression NF-κB p65 from 188.21% to 99.7% at 75mg/kg dose [106]. Ellagic acid showed a protective role against LPS-induced acute lung injury (ALI) in mice. The inhibition of the NF-κB pathway (reduction in TNF-α, IL-6, IL-1β secretions, and stimulated IL10 production) resulted in decreasing the pulmonary inflammation induced by LPS [107]. The pomegranate seed oil, flower and peel extracts were evaluated for adiposity, lipid profile, glucose homeostasis, and inflammation in high fat and high sucrose diet-induced obesity in mice. The combined treatment given to the obese mice with pomegranate seed oil (2mL/kg/day) and rosiglitazone (3mg/kg/day) for two weeks did not improve the glucose tolerance, but it reduced the fasting sugar blood glucose after four weeks of the treatment. The combination of pomegranate seed oil and rosiglitazone also improved insulin sensitivity. The extract of seed, peel, and flower further reduced the levels of proinflammatory cytokines IL-6, TNF-α and increased the level of antiinflammatory cytokines IL-10 [108]. Pomegranate extract rich in ellagitannins decreased the intestinal inflammation and proinflammatory cytokines in mucosa and serum. The pomegranate extract decreased p70S6K and genes Rp6ka2, Map2k2, and Mapk1 and sfn mRNA. The ellagic acid showed effective bindings against both IGF-1R mRNA and EGFR in both LPS-treated CCD-18Co cells (in vitro studies) and molecular docking studies. It reduced the expression of IGF-1R by 46% at 10mg/L GAE and 27% reduction with 4mg/L ellagic acid. Similarly, the EGFR mRNA was suppressed by 31% at 10mg/L GAE and 27% at 4mg/L of ellagic acid. The pomegranate polyphenols downregulated the mTOR which in turn led to the reduction in ERK1/2. Activation of AKT/mTOR pathways plays a role in cell proliferation and inflammation [109]. In another study, Kim et al. showed the antiinflammatory potential of the pomegranate extract in the dextran sodium sulfate (DSS)-induced colitis in SD rats and in vitro studies in LPS-treated CCD-18Co colon-myofibroblastic cells. Rats were fed with pomegranate extract with ellagic acid and ellagitannin protected against DSS-induced colon inflammation and ulceration. It also led to decrease in proinflammatory markers such as TNF-α and IL-1β, iNOS, and COX-2. Further, it reduced the phosphorylation activation of p70S6K1 and HIF-1α by 53% and 44% at 5 and 10mg GAE/L (markers of proliferation and inflammation). It also reduced the protein expression of p70S6K1 and H1F1α in LPS-treated CCD-18 cells. The ellagic acid suppressed the p70S6K1 and H1F1α mRNA by 62% and 68% at 5mg/L. The pomegranate extract induced the expression of miR-145 in LPS-treated cells in a dose-dependent manner. The cells were also treated with antagomiR which reduced the expression of miR-145 (reduces by 74%), and the pomegranate extract treatment 10mg GAE/L reversed the effect of miR-145 antagomiR (to 144%) [110]. The pomegranate leaf hydroalcoholic extract decreased the TNF-α mRNA and the protein levels (57% and 48% mean reduction) as compared to diclofenac (41% and 33%). Peritoneal leukocytes (neutrophils) were also reduced in the pomegranate extract-treated rat groups with acute peritonitis [45]. The same group also evaluated the antiinflammatory action of ethyl acetate fraction of the leaf extract of the P. granatum against LPS-induced acute peritonitis. The leaf extract-treated group (100–300mg/kg) showed reduced cell accumulation and collagen deposition in ALI. The inflammatory mediators such as lungs and bronchoalveolar lavage fluid, BALF TNF-α and albumin showed reduced expression 219.8±6.4pg/mL and 120.6±42.8pg/ml as compared to control vehicle (397±109.6 and 228.6±41.6pg/mL) when leaf extract (100mg/kg) and dexamethasone (5mg/kg) were given in lung tissue of mice. The 30mg/kg was not effective in reducing TNF-α, IL-1β, and IL-6 mRNA expression, whereas at a concentration of 100 and 300mg/kg downregulated the TNF-α and IL-1β but not IL-6. In the cultured LPS-treated RAW 264.7 macrophages showed reduced NO production and cytokines gene expression when incubated with leaf extract and kaempferol (100μg/mL) [46]. Shah et al. evaluated the effect of the P. granatum juice and punicalagin against nuclear factor kappa B (NF-κB) modulation of inflammatory bowel disease (IBD). The NF-κB is involved in the signaling pathway leading to the colon inflammation. The juice (400mg/kg p.o.) and punicalagin (4mg/kg) were fed to male SD rats. The colitis was induced in the animals with 2,4- dinitrobenzene sulfonic acid and on 18th day the colon sample was investigated for biochemical parameters (MPO, MDA, SOD, and NO) and cytokines (TNF-α, IL-1β, IL-18, and NF-κB). The pomegranate juice and punicalagin-treated groups showed decreased mRNA of the cytokines. The activity of NF-κB expression was decreased to 84% and 64% with pomegranate juice and punicalagin-fed groups. The phytochemicals punicalagin was suggested as a potential therapy for IBD [111]. Freeze-dried pomegranate fruit juice was studied for inhibiting neuroinflammation by evaluating PGE_2 production in IL-1β-stimulated SK-N-SH cells. The levels of COX-2, phospho-1kB, and phospho-IKK protein were evaluated. The NF-κB gene assay was carried out in TNF-α-stimulated HEK293 cells. The IL-1β-stimulated cells were also checked for the levels of BACE-1 and Aβ expression. The pomegranate extract in a concentration of 25 to 250μg/mL reduced the COX-2-dependent PGE_2 production. The inflammatory pathways phosphorylation of the 1kB and IKK were also reduced. Also, it inhibited NF-κB transactivation in TNF-α-activated HEK293 cells. The IL-1β-treated cells also showed decreased BACE-1 and Aβ levels in SK-N-SH cells [112]. The pomegranate flower extract at a concentration of 10–100μg/mL has been reported to decrease the production of NO (IC_50
=31.8μg/mL), PGE_2 (IC_50
=54.5μg/mL), IL6 (IC_50 value=48.7μg/mL), IL-1β (IC_50
=71.3μg/mL), and TNF-α (IC_50
=62.5μg/mL). The extract further inhibited the phosphorylation of the p38, JNK, ERK1/2, and translocation of the NF-κB p65 into nuclei [74]. The novel compound punicagranine has antiinflammatory potential. The compound showed the inhibitory activity against NO production with an IC_50 value of 22.8±1.2μM as compared to positive control hydrocortisone (IC_50
=3.76±0.49μM). The positive part is that the compounds at maximum concentration of 100μM against RAW264.7 cells do not show any cytotoxicity, which confirmed that the antiinflammatory activity is not because of the cytotoxicity [51].

11.6.9. Antidiabetic activity

Rouhi et al. evaluated the antidiabetic effect of pomegranate juice versus seeds powder and studied the effect on the lipid profiles, inflammatory biomarkers, levels of plasma glucose, and insulin. Diabetes was induced to 40 male SD rats by single intraperitoneal administration of streptozotocin (STZ; 60mg/kg b.w.) and nicotinamide (NAD; 120mg/kg b.w.). The diabetic rats were fed with 1mL of pomegranate juice and 100mg of pomegranate seed powder and then on 21st day 5mg/kg b.w. of glibenclamide. On 22nd day, the results showed that the pomegranate juice and the seed powder group did not show significant glucose reduction and levels of plasma insulin. The pomegranate juice reduced the total cholesterol and triglycerides and LDL nonsignificantly. However, pomegranate seeds significantly raise the plasma total cholesterol, LDL, and HDLs. Further, the plasma inflammatory biomarkers (NF-κB, TNF-α, and IL-6) were significantly reduced in both cases of pomegranate seeds and juice. Pomegranate juice (IC_50 value of 65.12±1.44μg/mL) showed 2.5 times more antioxidant effects as compared to the pomegranate seeds (IC_50
=159.90±6.24μg/mL) against DPPH radicals [63]. The isomers oleanolic acid 3-monodesmosides momordin and oleanolic acid 3-O-glucuronide reduced the blood glucose levels by slowing the gastric emptying. The presence of gallic acid (3,4,5-trihydroxybenzoic acid) and its dimers ellagic acid in the peel of the pomegranate can induce PPAR-γ expression and showed the hypoglycemic effects [72]. The aqueous-ethanol pomegranate peel extract inhibited the interleukin-1L-17 against the in vitro test of gut-associated lymphoid tissue. The extract also reduced the interleukin production in experimental animals with autoimmune encephalomyelitis. It downregulated the STZ-induced type 1 diabetes in C57BL/6 mice and experimental autoimmune encephalomyelitis female DA rats. The type 1 diabetes treated with pomegranate peel extract showed inhibition of immune cell infiltration into pancreatic islets and attenuated the production of IL-17 and IFN-γ in the gut-associated lymphoid tissue [56].

11.6.10. Pregnancy complication treatment

The healthy human placenta is necessary for successful pregnancy. Studies showed that oxidative stress led to dysfunction placenta and pregnancy issues. Pomegranate juice and punicalagin showed decrease in trophoblast injury in vivo and in vitro. The study was conducted in 12 pregnant patients. Four patients received 8oz./day of pomegranate juice, and eight patients were given apple juice (placebo). There was a significant decrease in oxidative stress in human placental tissue with patients who labored on pomegranate juice than on apple juice. Also, pomegranate juice reduced the apoptosis, oxidative stress, and cell death in hypoxia exposed to villous explants and primary trophoblast cultures. The polyphenol punicalagin showed more pronounced stimulus-induced apoptosis as compared to other polyphenol ellagic acid in cultured syncytiotrophoblasts [113].

11.6.11. Antilice and antidandruff activity

The aqueous and alcoholic extracts of P. granatum exhibited potent antilice and antidandruff activity. The hair growth potential was noted with increase in percentage of transition from the telogen to anagen phase as compared to minoxidil. The main phytoconstituent in the alcoholic extract was found to be maltol [114].

11.6.12. Antiacnes activity

Pomegranate extract can reduce the Propionibacterium acnes-induced edema in Wistar rat ears. Further, it was also able to inhibit the bacterial growth and activity of the lipase enzyme. The compound punicalagin and punicalin produced their effect as antibacterial, whereas granatin B inhibited prostaglandin E_2 production via decrease in COX-2 expression tested in heat-killed P. acne-treated RAW 246.7 cells. The compound punicalagin, strictinin A, and granatin B showed antilipase activities. The MIC and MBC of pomegranate extract against S. aureus and P. acnes was 62.5μg/mL. The MIC and MBC against P. acnes and S. aureus of punicalagin and punicalin were 6.25μg/mL (1.3μM) and 12.5μg/mL (2.2μM) [64].

11.6.13. Nephroprotective activity

Punicalagin and ellagic acid have been studied for their protective roles on the kidney with induced focal segmental glomerulosclerosis in rat through nephrectomy and injecting adrenaline. The pretreatment with P. granatum tannins and benazepril showed improvement in renal pathology and reduced expression of PDGF, α-SMA, and Ang-II. Further the tannin extracts normalized expression of TGF-β1/Smad [55]. The methanolic extract of P. granatum leaves showed promising protective effects against oxidative stress of gentamicin-induced nephrotoxicity. It was observed that coadministration of the methanol extract leaf of P. granatum and gentamicin protected the kidney from the nephrotoxicity induced by gentamicin in rats. The extract-treated groups showed decrease in TNF-α in a dose-dependent manner. Renal function was normalized in the treated groups with a decrease in albumin excretion, serum creatinine, and blood urea nitrogen levels. The group also showed decrease lipid peroxidation levels and improved renal morphology, severity of edema, mononuclear infiltration, and vacuolar degeneration [115].

11.6.14. Cardioprotective activity

Razani et al. evaluated the cardioprotective effect of pomegranate juice in patient with ischemic heart disease. The study included 100 patients diagnosed with unstable angina and myocardial infarction. The patients were given 220mL pomegranate juice daily along with the conventional therapies. The levels of serum troponin and MDA showed significant decrease with reduction in the intensity and duration of angina pectoris. The protein troponin is the major protein present in cardiomyocytes and important marker which gets leaked from damaged cells [65]. The compound punicic acid also works as hypoglycemic agents acting on PPARγ which also inactivate the proinflammatory mediators such as NF-κB and TNF-α [116]. A clinical study evaluated the antioxidant effects of the pomegranate juice on cigarette smoking-induced cardiac hypertrophy, vascular inflammation, and injury. The pomegranate juice given to the cigarette smoke group of rats having been exposed to cigarette smoke for four weeks (five days in a week) reduced cardiac hypertrophy and lessened the atherosclerosis markers. It also prevented aortic calcification and reduces the inflammatory mediators (bradykinin 1 and 2 receptors, IL-1β, and TNF-α). Pomegranate juice also reduced the posthypertrophic and oxidative stress markers (fibrotic/atherogenic-related markers Fn1 and ObR) [117].

11.6.15. Hepatoprotective activity

The effect of pomegranate juice on prevention of the nonalcoholic fatty liver disease/nonalcoholic steatohepatitis was evaluated by Noori et al. SD rats were fed with high sugar and high fat and pomegranate juice for seven weeks. The pomegranate juice animal group showed lower hepatic steatosis, lower hepatic proinflammatory cytokines (TNF-α, IL-1β, and IL-6) and profibrotic gene expression, lower glucose, ALT, AST, insulin, and inflammation (lobular and portal). The proinflammatory cytokines TNF-α involved in inflammation, insulin resistance, and NAFLAD [118].

11.6.16. Miscellaneous activities

The flower extract of P. granatum was evaluated for minimizing the peritoneal adhesion after abdominopelvic surgery. It was found that 400mg/kg/day of the flower extract for eight days showed less adhesion with lower rate of foreign body reaction in the serosal layer as compared to control [119]. Wang et al. described the vasculoprotective effect of the pomegranate due to the presence of hydrolysable tannins, ellagitannin, and ellagic acid. It also showed the presence of anthocyanins and flavonoids. These phytoconstituents were responsible for reducing the platelet aggregation, oxidative stress, lipid uptake by macrophages, thereby improving the endothelial function. This further regulated the blood pressure and lessened the chances of atherosclerosis and hypertension [28].

11.7. Role of phytoconstituents in the management of environment pollutant

The peel extract of P. granatum was used to develop Ag/Fe_3O_4/RGO nanocomposite (NC) for the reduction of organic pollutants such as 4-nitrophenol, methylene blue, and methyl orange in water at mild conditions. The major polyphenol components present in the P. granatum extract such as ellagic tannins, ellagic acid, and gallic acid act as reducing and protecting agents for the reduction of metal ions and stabilize the nanoparticles. The DPPH radical scavenging studies confirmed the presence of total phenolic components which were able to reduce the graphene oxide and silver ions to form silver nanoparticles. The results showed highly effective catalytic activity of the NCs, i.e., synergistic effect of Ag nanoparticles immobilized over reduced graphene oxide (RGO) and Fe_3O_4 magnetic composite against the reduction of different dyes in few seconds [120]. Orange G dye present in textile wastewater was degraded up to 61.45% with the laccase activity. This was increased in the presence of redox potential of the P. granatum peel extract up to 85% and with certain conditions to 97%. The toxicity was measured in case of germination of Trigonella foenum-graecum L seed with the treated and untreated dye. It was noted that the seed germination was completely inhibited (100%) with untreated dye solution and 81.03% growth with treated dye solutions [121]. P. granatum peel biochar (obtained by pyrolyzed at 300 and 600°C) was evaluated for Cu (II) absorption. The quality of the biochar developed at 300°C had less ash content, surface area, pH, and cation exchange capacity than biochar 600°C. Biochar produced at lower temperature showed higher organic carbon contents and decreases with increasing the temperature. There was an increase in the soil pH because of the alkaline pH of the biochar due to the presence of negatively charged carboxyl, hydroxyl, and phenolic groups. The biochar showed nearly double adsorption capacity as compared to the original soil. The adsorption isotherm showed monolayer adsorption of Cu (II) on biochar [122]. P. granatum fruit rind fractions and the compounds present were found to be helpful and may resolve the problems associated with the synthetic pesticides. The purified column fractions were evaluated against Microcerotermes beesoni termite. The column fraction of the pomegranate was significantly effective as compared to standard 20% chlorpyriphos because of the higher concentration of quercetin [54]. The P. granatum peel extract of ZnO with bovine bone-derived hydroxyapatite (ZnO/HAP) in the shape of cauliflower was synthesized by Sin et al. The ZnO/HAP-5 composite/photocatalyst demonstrated the highest antibacterial activity against Enterococcus faecalis. The mechanism of action was due to the morphological and cell alteration to the bacteria leading to the K+ and protein leakage. The antibiofilm performance of the composite was checked against E. faecalis and found to be excellent by reduction in exopolysaccharide and hydrophobicity index. The photocatalytic improvement in activity was because of efficient charge separation and high H_2O_2 generation. The composite also showed antioxidant activity inhibiting activity of DPPH-free radicals. The photocatalyst may be utilized for safe and practical water disinfection [123].

The orally administered hydroethanolic extract of P. granatum fruit peel was evaluated for genotoxic and cytotoxic effect in mice. The extract reduced the frequency of micronucleus showing potent antigenotoxic effect up to 82.8% after 24h and 90.6% after 48h. Also, no subacute toxicity was noticed at 700mg/kg of the dose. A low toxicity was noted in histopathology of the spleen, heart, and kidney after long-term administration of the fruit peel extract. The fruit peel extract can be used as an alternative therapy to other chemicals in preventing oxidative damage [124]. The production of Dy_2Ce_2O_7 nanostructures was developed taking metal nitrates and juice of P. granatum as fuel. The nanostructures were developed by applying the juice of P. granatum as fuel at 450°C for 4h showed excellent photocatalytic activity. The nanostructure was found to be efficient in destruction of the erythrosine and methylene blue pollutant photocatalytically [125].

11.8. Applications of P. granatum in nanotechnology

The nickel and silver nanoparticles on RGO composite were developed by Das et al. with the help of P. granatum juice. The developed electrode showed greater electroactive areas, roughness factor, and had lower electrode resistance (30 Ohm). The electrode was successfully used for detecting ascorbic acid electrochemically in the linear range (4.89–90.09μM), limit of detection (0.16μM), and sensitivity (23,381.8μAcm
−2 mM
−1). The electrode had excellent stability for up to 45 days [126]. The bioactive phenolic compounds present in hydroalcoholic (70% methanol) crude extract and the methanolic fraction of P. granatum peel were used to reduce the Ag
+ to Ag(0). Esawy et al., developed the silver nanoparticles mouthwashes from crude and methanolic extracts [127]. Similarly, the reduction and stabilization of the zinc-to-zinc oxide nanoparticles were developed using an environmentally friendly P. granatum fruit peel extract at higher annealing temperature of 600°C (32.98nm) and 700°C (81.84nm). The synthesized nanoparticles were very effective in inhibiting the growth of E. coli and E. faecalis. The MIC value of the nanoparticles developed at 600 and 700°C against E. coli was 64.53 and 90.90μg/mL whereas against E. faecalis was 22.09 and 95.21μg/mL. The nanoparticles were equally cytotoxic against human colon normal and cancerous cells at concentration of ≥31.25μg/mL [128]. Gold nanoparticles were developed with the help of reducing potential of fleshy pulp (mesocarp) of pomegranate. The reducing potential of pomegranate is because of the phytoconstituents such as amino acids, polysaccharides, phenolic acids, flavonoids, tannins, and anthocyanin [129]. The peel extract of P. granatum was used to form copper oxide nanoparticles by highly efficient sol–gel method. The synthesis was performed after the reaction of aqueous solution of P. granatum peel extract and copper acetate monohydrate [130]. The peel extract of the P. granatum was used to develop the magnetic iron oxide nanoparticles and was evaluated for anticancer activities. The magnetic iron oxide nanoparticles did not show any reactivity against breast (MCF7), colon (HCT116), cervical (HeLa), and lungs (A549) cancer cell lines and the IC_50
>250μg/mL, but when combined with 2% and 4% of peel extract showed potent anticancer activities (IC_50 197.46 and 85.06) against nasopharyngeal carcinoma (HONE1) cell line [131].

11.9. Industrial uses of pomegranate

The nonedible part of the pomegranate was in vitro digested and fermented simulating a large intestinal process and change in the phenolic bioaccessibility. The untargeted metabolomic resulted in the transformation of flavonoids by in vitro fermentation. Untargeted metabolomic used as a tool to check the chance of the phenolics in the areas of the gut. There was increase in percentage bioaccessibility of phenolic acids and tyrosols in flowers (maximum after 24h) due to insoluble dietary fiber contents. The results supported the fact that nonedible part of the P. granatum can be used for value-added food [132].

P. granatum juice appeared to be a good carrier of the probiotic especially Lactobacillus species (Lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus salivarius, and Lactobacillus casei). The phytoconstituents and nutritive value of the juice fulfill the needs of the bacteria for fermentation. This probiotication further maintains the antioxidant value of the pomegranate juice. The antioxidant effects are because of the presence of the phenolic acids (rosmarinic acid and caftaric acid) and flavonoids (rutin, quercetin, quercetin-3-glucoside, kaempferol rutinoside). The optimum temperature for the growth of bacteria was found to be 37°C and pH 4. This is helpful for people who develop lactose intolerance to dairy products [133]. The P. granatum fruit (peels and membrane) are promising source of pectic polysaccharide. The polysaccharide obtained contain 76% uronic acids after extraction with aqueous solution of ammonium oxalate (NH_4)2C_2O_4. The polysaccharide contains high methyl esterified and low acetylated pectin. It also has cross links of glucuronoxylans and xyloglucans polysaccharides that confirm close interaction between them in the cell walls [134]. The seed juice by-product of P. granatum was added to the fish gelatin reinforcing the antimicrobial agents for eco-friendly packaging of food. The seed juice extract mainly contains xylan and cellulose as the main components. It also contains high total phenolic compounds and possesses antioxidant activity. The 10% wt. of seed juice by-product of P. granatum showed the best performance. There was no barrier difference for oxygen, and water vapor penetration permeability was noted as compared to control. The developed film also showed high thermal stability along with water resistance and inhibitory effect against S. aureus, thereby protecting the food from lipid oxidation [135]. Wang et al. improved the oxidative stability and sensory properties of the sunflower oil flavored with P. granatum cv. Heyinshiliu. A total of 21 compounds were isolated, among them the major components consist of methyl palmitate (22.2%), myrcene (18.8%), and 2,6,11,15-tetramethyl hexadecane (8.7%). The oil becomes flavored when an essential oil from the P. granatum was mixed with sunflower oil. This further increased the acidity value, peroxide value, p-anisidine value, and total oxidation value on accelerated storage of sunflower oil at 65°C for 30 days. The change in fatty acid components along with the thiobarbituric acid reactive substance and total polar compounds (altered triglycerides) were inhibited. The addition of essential oil further improved the taste, flavor, and appearance [136].

11.10. Patents granted on P. granatum

P. granatum is well explored for its bioactive metabolites and their uses in various pharmacological conditions. Most of the studies were focused on the enhancement of human health, and during this, many international patents have been granted to various research groups. Number of reports are published describing the importance of P. granatum in human health enhancement and patent details [137–140]. We have compiled some of the patents granted to P. granatum in Table 11.3.

Table 11.3

Some of recently granted patents to Punica granatum and its preparations (2009 onwards).

S. No. | Patent title | Inventors | Patent app no. | Year | Ref

1.

Herbal composition used for waist and tummy firming has extracts and oils of Cyprus, Ajowain, lemon, black pepper, pomegranate, rosemary, Geranium, majuphal, doorva grass, gehun ankur, spirulina, and soybean in specific weigh percentage

M. I. Khan, A. H. Zaidi.

IN200800662-I1

2009

[141]

2.

Hydrating mist formulation, useful in personal care for, e.g., improving and retaining the moisture level in the skin, comprises, e.g., pure rain water, Hamamelis virginiana, menthyl lactate, natural preservatives, and aroma imparting oils.

P. Vilinsky

WO2010022393-A1

2010

[142]

3.

Skin external composition useful for preventing skin dryness and skin aging, comprises extracts of Punica granatum and Tussilago farfara

G. Cho, D. Kim, E. Kim, H. Kim, S. Kim, E. Moon, H. Noh, C. Park

KR2010031839-A

2010

[143]

4.

Cosmetic use of an active agent mixture, obtained from, e.g., Clintonia borealis or P. granatum, for preventing and/or inhibiting the effect (i.e., nonpathological effect) of psychoemotional stress on the hair

M. Briese, R. Ghosh, Y. Oezka, T. Weiss

DE102009043486-A1

2010

[144]

5.

Antioxidant composition, useful for improving the appearance of the skin, comprises agent derived from Lycium barbarum, P. granatum, Vitis vinifera, Aspalathus linearis, and Camellia sinensis, and vehicle, e.g., glycerine

J. Jacobs

US2010119463-A1; WO2010056675-A2; WO2010056675-A3

2010

[145]

6. | Skin care formulations | D. Burke-Colvin, M. Hines, D. Gan | US8048456B2 | 2011 | [146]
7. | Use of an extract of P. granatum for combating canities | S. Commo, S.A. Nestec, S.A. LOreal | US2012/0263812 A1 | 2012 | [147]
Table Continued

S. No. | Patent title | Inventors | Patent app no. | Year | Ref

8.

Dietary food supplement containing natural cyclooxygenase inhibitors and methods for inhibiting pain and inflammation

M.G. Nair, D.L. Dewitt, H. Wang, D.W. Krempin, D.K. Mody, D.G. Groh, M.A. Murray, M. Lemay, Y. Qian, A.J. Davies, R. Dykhouse,

US 8,337,91

2012

[148]

9. | Antiangiogenic extracts from pomegranate | F.L. Greenway, Z. Liu, E.A. Woltering, | US 8,334,000 | 2012 | [149]
10. | Pomegranate extract for use in treating rhinitis and sinusitis | E.H. Budman, A. Yaari, I. Braverman | US2014/0377389 A1 | 2014 | [150]
11. | Therapeutic composition produced using P. granatum and hydrogen peroxide | A.L. Huang, G. Wu | US8343552 | 2013 | [151]
12. | Pomegranate extract for use in treating rhinitis and sinusitis | E.H. Budman, A. Yaari, I. Braverman | US 14/382,867 | 2014 | [150]

13.

Products for oral administration comprising extracts of P. granatum (pomegranate), intended for a pet, and applications of same

P. Monginoux, H. Gatto, C. Karst, F.R. Waldenberger

US2014/0335180 A1

2014

[152]

14. | Pomegranate extract having a high ellagic acid content and use of the pomegranate extract | H.Y. Lee | US8986749 | 2015 | [153]
15. | Pomegranate-derived cell culture and methods for preparing and using the same | Y. Hagay, M. Azachi, R. Yatuv, | WO2015102003 | 2015 | [154]

16.

Antiresorptive and bone-building dietary supplements and methods of use

D.W. Krempin, M.A. Murray, Y. Lin, K.W. Gellenbeck, S. R. Da Costa, L. M. Wilkins, H. Roh-Schmidt, J. Rana, J. F. Rebhun, D. J. Fast.

US 9,446,087

2016

[155]

17. | Formulations containing pomegranate seed oil, Rosa canina fruit oil, and Inula viscosa oleoresin or extract | T. Sofer, T. Meyer | US 15/521,978 | 2016 | [156]
18. | Lip gloss | K. Wilson | US 9,295,635 | 2016 | [157]
19. | Novel antiwrinkle and antiaging nanoformulations and method of preparation using novel nano codelivery system | A.I. Orza | US 15/334,310 | 2017 | [158]

11.11. Pomegranate in clinical trials

The double-blind clinical trial was carried out in 34 chronic gingivitis patients for four weeks with 5% pomegranate peel extract. The results of German clinical trial (DRKS-ID: DRKS00010602) showed no adverse effect and was found a promising alternative in managing chronic gingivitis [159]. In a double-blind clinical trial, P. granatum mucoadhesive gel was evaluated against oral recurrent stomatitis and compared with Triadent oral paste. The study recruited 60 patients, and time of pain elimination and complete healing was noted. The Kaplan–Meier analysis revealed that pain relief time in the case of P. granatum was less than placebo group, whereas there was no difference in pain relief time in case of Triadent group and placebo. The wound healing was also lower in the case of P. granatum as compared to the Triadent and placebo group [160]. Another double-blind trial carried out by Goshtasebi et al. showed the P. granatum flower to possess antihemorrhagic activity [161]. A phase II clinical trial studied the efficacy of pomegranate in male patients with prostate cancer with rising prostate-specific antigen (PSA) after first therapy. The patients were selected with rising PSA (>0.2 and <5ng/mL) after surgery and radiotherapy. The patients were given eight ounces of pomegranate juice daily, a total of 570mg polyphenol GAE. The study reported no major adverse event, and the trial results were encouraging. The growth of LNCaP, apoptosis, and serum nitric oxide in patient serum was compared pretreatment and posttreatment. The result showed that LNCaP cell proliferation, reduced by 12% with 17% increase in apoptosis and 23% increase in serum nitric oxide. It also showed reduction in the sensitivity to oxidation of serum lipids and reduction in oxidative state [162]. A recent clinical trial (randomized, placebo-controlled, double-blind, parallel 2-arm 1:1) was conducted in the hospitalized mild to moderate coronavirus disease patients on inflammatory biomarkers, C-reactive protein (CRP), interleukin 6 (IL6), erythrocyte sedimentation rate, and complete blood count [163]. The result of the clinical trials is not yet out.

A 12-month randomized, double-blind, placebo-controlled study was conducted among the old and middle-aged groups (261 aged group 50–75yrs) for the effect of pomegranate on the memory. Memory was measured by brief visuospatial memory test-revised (BVMT-R) and Buschke selective reminding test (SRT) at interval of 6 and 12 months. Baseline variables between the tested and the placebo group did not differ significantly. It was concluded that daily consumption of juice of P. granatum may stabilize the ability of learning and visual information [164]. Another study on 32 subjects (older with age-associated memory complaints) through placebo-controlled, randomized trial showed significant improvement in Buschke SRT of verbal memory after taking eight ounces of pomegranate juice for four weeks. The group also showed increased plasma Trolox-equivalent antioxidant capacity and had increased functional brain activation (fMRI) in verbal and visual memory task as compared to the placebo control [165]. A parallel block-randomized clinical trials (RCTs) were carried out in 163 patients with pomegranate polyphenols to evaluate whether it enhances cognitive function after stroke. Pomegranate polyphenols pills (1gm of polyphenols or equivalent to 8 oz. of juice) and placebo twice a week after two weeks of stroke. Results showed neurological and functional improvements in patient under pomegranate pills and patients spent less time in the hospital as compared to the placebo controls [166]. Another single-blind, RCT for the effects of pomegranate juice consumption on blood pressure and lipid profile in patient with type 2 diabetes was conducted. A total of 60 subjects were taken for this study (30 intervention and 30 control) and were given 200mL/day of the pomegranate juice for six weeks. The measurement of the systolic blood pressure, diastolic blood pressure, total cholesterol, LDL, HDL, and triglycerides was measured at interval of 12–14h of fasting (baseline) and at the end of the study. After six weeks, systolic (13.5±1.5mmHg vs. 12.3±2.5, P
<.001) and diastolic blood pressure (7.7±1.6 vs. 7.2±1.6mmHg, P
<.05) were significantly decreased in intervention group as compared to the control. There was no significant change in the cholesterol levels except that total cholesterol and LDL cholesterol decreased significantly [167]. Another RCT evaluated the short-term effect of fresh pomegranate juice on serum cortisol and thyroxine in patients with type 2 diabetes. The study included 89 fasted patients with type 2 diabetes and were given fresh pomegranate juice at a dose of 1.5mL/kg. Blood was collected after 1 and 3h of juice administration. It showed that the level of serum cortisol decreased but no effect on thyroxine levels [168]. In another RCT, 49 overweight-obese with mild hyperlipidemia were given two doses, i.e., first and second doses of pomegranate extract or placebo for three weeks. The effect showed decrease in plasma LPS-binding protein and small decrease in high-sensitive CRP (after second dose of 656mg phenolic). These LPS markers associated with gut microbiota are indicators of endotoxemia in overweight people. Pomegranate extract also led to a decrease in the proinflammatory microorganism such as Parvi monas, Methanobrevi bacter, and Methanosphaera and increase in Faecali bacterium and Odori bacter that maintains a normal balance of gut microbiota [169]. Another detailed data from the 24 clinical studies on pomegranate extract showed no statistical significance for the prevention and treatment of cardiovascular disease. The more confirmatory test and the dose findings are therefore required to prove the clinical benefits of the pomegranate extract in preventing or treating the cardiovascular disease [170]. The data of some RCTs for the effects of pomegranate juice and blood pressure were studied and was found that 8 RCTs showed significant reduction of both systolic and diastolic blood pressures. It was further observed that pomegranate juice reduced the systolic blood pressure regardless of the dose and duration (>12weeks, weight mean difference=
−4.36mm Hg and <12weeks, weight means difference=
−5.83mm Hg) (>240cc weight mean difference=
−3.62 and<240cc weight mean diff. −11.01), whereas borderline effectiveness in reducing diastolic blood pressure was observed for dose of >240cc [171].

11.12. Toxicity of pomegranate

There is a scarcity of data on the toxicity of pomegranate. It was observed in Victoria Australia that large amount of the pomegranate fruit feed to the cattle led to the death of the several cattle. The bioassay-mediated approach was used to identify the metabolites responsible for the cause. It was found the presence of gallic acid, ellagic acid, and punicalagin. Only a small amount of gallic acid was detected, but it was found that in the rumen of the cattle, microbial metabolism of the punicalagin results in the conversion to gallic acid contributing to the toxicity [172].

11.13. Conclusion

The chapter outlines nutritional, medicinal, and economical importance of P. granatum. Various phytochemical studies have indicated the presence of different phytoconstituents present in juice, peel, seeds, leaf, and flowers. The secondary metabolites of pomegranate are mainly responsible for its wide spectrum of pharmacological activities. The plant part extracts and isolated natural products showed inhibitory effects against cancer, inflammation, leishmaniasis, viruses, and neurological disorders.
Other positive effects of pomegranate extract supplementation include antidiabetic, antiacne, antioxidant, nephroprotective, cardioprotective, hepatoprotective, and lipid-lowering activity. One of the pomegranate polyphenols supplementations, especially the ellagic acid, acts as SERMs in the treatment of breast cancer. Ellagic acid being flat can act as intercalating agent and binds with DNA but needs more clear understanding. The presence of high phenolic content such as ellagitannin (punicalagin, punicalin) and the flavonoids (quercetin) in the peel of pomegranate can accelerate the recovery from neuroinflammation and oxidative damages in neurodegenerative diseases. The ellagic acid and punicalagin present in the peel showed greater virucidal and antileishmanial activity. The chapter highlights the pomegranate juice has advantageous effects on the heart, liver, and kidney, but the supporting cellular mechanisms for the activity are insufficient. Additionally, the phenolic compounds present have not only wide application in the formulation of nanoparticles and management of environment pollutants but also used industrially as value-added foods. The pomegranate extract showed no evidence of toxicity or subacute toxicity up to 700mg/kg. However, long-term pomegranate peel extract use may cause low toxicity as recovered by histopathology of heart, spleen, and kidney in experimental animals. Therefore, more data through research are required to optimize pomegranate supplementation in terms of its efficacy versus toxicity in humans.

List of. abbreviation

AIDS    acquired immunodeficiency syndrome
ALT    alanine aminotransferase
AST    aspartate transaminase
BDNF    brain-derived neurotropic factor
CAM    chorioallantoic membrane
COX    cyclooxygenase

CREB    cyclic AMP-response element binding protein

DMBA
7,12-dimethylbenz(a)anthracene

DPPH
2,2-diphenyl-1-picrylhydrazyl

DSS    dextran sodium sulfate
EGCG    epigallocatechin gallate
EGFR    epidermal growth factor receptor
FAB    fast atom bombardment
FGF2    fibroblast growth factor
GAE    gallic acid equivalent
GC-MS    gas chromatography–mass spectrometry
HDL    high-density lipoprotein
HHV    human herpesvirus
HIF-1α    hypoxia-inducible factor-1α

HPLC    high-performance liquid chromatography

HSV    herpes simplex virus

IL-6    interleukin 6

LC-ESI-QTOF-MS/MS    high-performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry

LDL    low-density lipoprotein
LOX    lipoxygenase
LPS    lipopolysaccharide
MBC    minimum bactericidal concentration
MDA    malondialdehyde
MDR    multiple drug resistance
MIC    minimum inhibitory concentration
MMP-2    matrix metalloproteinase-2
MPO    myeloperoxidase
NAD    nicotinamide
NFAT    nuclear factor of activated T-cell
NMR    nuclear magnetic resonance
NO    nitric oxide
PDA    photodiode-array detection
PDGF    platelet-derived growth factor

PPAR-γ    peroxisome proliferator-activated receptor gamma

QE    quercetin equivalent
RCT    randomized clinical trials
SFE    supercritical fluid extraction
SERM    selective estrogen receptor modulator
SOD    superoxide dismutase
SWE    subcritical water extraction

TBARS    thiobarbituric acid reactive substance

TIMP-2    tissue inhibitor of metalloprotease-2

TNF-α    tumor necrosis factor alpha

VEGF    vascular endothelial growth factor

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Chapter 12: Garcinia cambogia

George Joseph, Akhila Nair, and Sasikumar Kuttappan
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Abstract

The demand of herbal products has seen an increase in their consumption in nutraceuticals and cosmetics world due to globalization. In association with this, Garcinia cambogia (syn.) (Guttiferae), the evergreen tree has gained much attention because of its active ingredient, hydroxycitric acid that is well documented for treating obesity. The fruit rind of Garcinia, is treasured among South Asia as a food preservative and flavoring agent from time immemorial. This culinary agent has other medicinal value to treat various diseases including constipation, rheumatism, irregular menstruation, piles, sores, dermatitis, hemorrhoids, dysentery, and edema. Besides, the adulterants that can be plausibly used should be identified and strict regulatory norms should enforce basic legislation for these ingredients to be used as supplements. Hence, the fruit of Garcinia holds a paramount value as nutraceutical and dietary supplements and should be considered for large-scale evidence-based studies.

Keywords

Hydroxycitric acid; Antiobesity; Garcinia cambogia; Weight loss

12.1. Introduction

Garcinia cambogia L. belongs to the family Guttiferae (Clusiaceae) and is native to Southeast Asia. It is mainly native to India, Nepal, and Sri Lanka and now it is distributed in Malaysia, China, and Philippines. In India, it is distributed over the Western Ghats around Maharashtra, Tamil Nadu, Kerala, and Karnataka. The tree of Garcinia is grown well over flooded or waterlogged soils near valleys, river banks, and can tolerate drought as well as undulating groundwater [1]. This tropical dicotyledonous tree is renowned as “Malabar Tamarind.” The tree achieves an approximate height of 20m with drooping, horizontal branches, and rounded crown. The leaves of the tree are shiny, ovate, and dark green in color (Fig. 12.1) and fruits are ovoid in shape with six to eight grooves bearing yellow, red, or purple color. The seeds of the fruit have succulent aril part circumference around it. Summer seasons are favorable for flowering, and rainy seasons are supportive for ripening of fruits. The active ingredient of Garcinia is the organic acid named hydroxycitric acid (HCA) [1,2 dihydroxy propane 1,2,3 tricarboxylic acid] followed by benzophenones including garcinol and guttiferones and xanthones [1]. Traditionally, the fruit is used in culinary, especially in fish curries and other food preparations in South India and Sri Lanka because of its sourness as well as astringent taste. It is used as a food preservative, astringent, and purgative. The fruit rind is known to treat numerous diseases including dysentery, bilious digestive conditions, intestinal parasites, angina, rickets, bone fractures, spleen enlargement, and tumors. Scientifically, HCA has gained acclamation as a dietary supplement as an efficient weight management product. Other components of garcinia especially garcinol and guttiferones have also registered to possess antioxidant, antidiabetic, antiulcerogenic, antiinflammatory, hepoprotective, cytotoxic, erythropoietic effect, etc. Howbeit, evidence-based studies to strongly anchor these effects are meagre [2].

12.2. Phytochemical analysis

G. cambogia has different phytochemical ingredients including xanthones and isoxanthohumols, such as garbogiol, rheediaxanthone, and oxy-guttiferones; benzophenones such as garcinol, isogarcinol, and guttiferones; and organic acids [1]. The organic acids (Fig. 12.2), which are isolated from this plant includes HCA, succinic acid, malic acid, citric acid, tartaric acid, and garcinia lactone. Among these, HCA is the major biologically active compound [2].

Figure 12.1
Photographs of the Garcinia cambogia tree in habitat, leaves, and dry fruits.

Figure 12.2
Chemical structures of organic acids isolated from Garcinia cambogia.

12.3. Hydroxycitric acid

HCA is a popular weight-controlling food additive that is obtained from the wilted fruit skin of a small pumpkin-shaped fruit of the tree G. cambogia. This skin of G. cambogia contains calcium or potassium salt of HCA [3]. HCA is the vital acid found in the product of G. cambogia. Although HCA can be secluded through various techniques in its free structure, as a calcium or potassium salt or as lactone, normal tamarind extracts are now retailed as calcium or potassium salts of HCA, containing roughly 60% HCA [4]. HCA was recognized by Watson and Lowenstein (1970) as a powerful aggressive inhibitor of the extramitochondrial protein adenosine triphosphate-citrate (pro-3S)-lyase, which is associated with the extramitochondrial combination of unsaturated fats [5,6]. HCA is water soluble and can be easily detected by Liquid Chromatography (LC) procedures. Most of LC techniques were portrayed estimating HCA in extracts and plans to control the nature of these items, deciding the amount of HCA [7,8]. Gas chromatography joined to mass spectrometry (GC-MS) have been depicted measuring HCA in human plasma [9,10]. Authentic HCA was prepared from G. cambogia, Hibiscus sabdariffa which is roselle plant. The methyl ester of HCA was identified by nuclear magnetic resonance analysis [7]. Higher plants and fungi have different routes for the biosynthesis of abscisic acid. Microorganisms that produce HCA provide a potential alternative source to hibiscus HCA [11]. Super CitriMax (HCA-SX), a well-liked dietary enhancement, is additionally gotten from the dried natural fruit skin of G. cambogia and comprises a calcium or potassium salt of HCA. The mutagenicity of HCA-SX was examined utilizing the Ames, chromosome deviation, and micronucleus tests [12].

12.4. Pharmacological activities and mechanism of action

Due to the notable elevation in the obesity rate, people's health has been facing in numerous nations of the world, one of the serious issues caused by this disorder. So, for the treatment of stoutness, the concentrate got from the dried products of G. cambogia, which has been considered and utilized as a characteristic enhancement for weight reduction [13]. The HCA is obtained from the skin of the fruit, which goes about as a likely enhancement to the weight control by causing the hunger concealment and decreasing the body's capacity to shape fat tissue [14]. G. cambogia extract and its dynamic part HCA have likewise been utilized to decrease cholesterol and fatty oils. It is additionally available with the option of calcium, magnesium, potassium, and combinations thereof because of different impacts, like the improvement of glucose resistance and of circulatory strain [15,16]. The investigation of Kovacs and Westertep–Plantenga closed, in a test condition in people, that the treatment with HCA during overloading with aldehyde utilitarian gathering (-CHO) can lessen the de novo lipogenesis [17]. In rats, Kim et al. [18,19] confirmed that the supplementation of this herbal medication assisted with diminishing the muscle versus fat, however, not to diminish weight or hunger. The hypothesis behind the G. cambogia/HCA is that it functions as an enemy of corpulence specialist by acting in the neuroendocrine pathways, connected with the satiety, creating an anorectic result, by advancing the restraint of citrate lyase enzyme that smothers the hunger and expands the cutting of fat in body. Subsequently, it helps with controlling hunger, with a resulting decrease of food consumption, caloric limitation, and weight reduction. By hindering the citrate lyase, the body expands the oxidation of carbohydrates and restrains the lipogenesis [14,20,21]. G. cambogia supplementation can be utilized as a metabolic controller of weight and lipid irregularities in warm blooded creatures. A study claimed that G. cambogia/HCA lessens the lipid levels in blood, like fatty oils and cholesterol, other than of expanding the thermogenesis [16]. The investigation of Amin et al. noticed constructive outcomes, like diminished hunger and improvement of the unsafe impacts brought about by high fat and sucrose consumes less calories in rats [22]. The investigation of Cheng et al. featured that the HCA supplementation upgraded the pace of glycogen union in human skeletal muscle and further developed the postmeal insulin responsiveness [23]. It likewise guaranteed that there aren't any known after effects for the use of this herb. Be that as it may, it is not suggested for individuals determined to have diabetes or individuals experiencing any sort of dementia or condition, including Alzheimer illness along with pregnant and lactating ladies and has contraindications in regards to the accompanying utilization of specific medications. There are many investigations that propose positive outcomes concerning the viability of the G. cambogia in the weight reduction process, by lessening the hunger, the level of fat, the lipogenesis interaction, as well as the enhancement of biochemical levels, such as fatty oils, cholesterol, and glucose, and muscle glycogen amalgamation [13]. G. cambogia and raspberry ketone (RK) are the nutraceutical agents that are broadly utilized over the counter to control weight [24]. RK is an aromatic compound of red raspberries and utilized in an assortment of items prompt weight reduction [25]. RK was demonstrated to upgrade fat tissue digestion and thwart the ingestion of dietary fats from the small digestive organs [26]. Studies show that using G. cambogia and RK single regimens decreased both serum insulin and glucose levels along with a decline in insulin resistance confirming the results of a previous work on RK against nonalcoholic steatohepatitis model. G. cambogia, RK, and their combination successfully induced weight loss, where combining both agents showed the highest slimming effect to concur with the results of Kim et al.; however, the impact of one or the other on leptin and insulin-signaling transduction pathway still needs to be clarified [27,28]. In a rat study, the rats fed huge fat fructose diet showed a notable elevation in body weight within one month of feeding, which is compared to the rats that fed a normal diet (ND). However, during the nursing period (two months), G. cambogia and RK regimens notably reduce the body weight, especially the combination regimen, as compared to the fatty nontreated rats [29]. Ultra-pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) bioanalytical technique was proficiently applied to the HCA pharmacokinetics study investigating more than plasma tests [30].

12.5. Common adulterants

The notable antiadiposity properties of G. cambogia, the market interest, and worth of its items are expanding step by step. Hence, it is thought that it very well may be tainted with some other Garcinia species, which could altogether lessen its quality potential. Consequently, it is vital to break down debasement in its items, and to foster protected and fast logical techniques which could confirm G. cambogia items [31]. In India, Garcinia indica is utilized for flavourful for curry and furthermore in customary medication [32]. G. indica natural product is broadly produced in India and is widely supplied locally as well as internationally. Thus, because of the huge availability and simple openness of G. indica, it is thought that G. cambogia items could contain defilements from G. indica. Studies show that G. cambogia and G. indica organic products contain xanthochymol, isoxanthochymol, and HCAs [33]. The phytochemical investigations of G. indica have shown that the product skin contains elevated content of anthocyanins [34,35]. Anthocyanins also have some potent biological activities [36]. In this manner, anthocyanins could be a decent choice to examine the defilement in G. cambogia items. The adulteration in G. cambogia items was dissected utilizing high-pressure liquid chromatography and exceptionally delicate technique for ultra-pressure liquid chromatography combined with an electrospray ionization time-of-flight mass spectrometry (ESI-IT-TOF-MS) [37]. A common issue in herbal products is corruption in high worth crude medications and completed items [38–40]. The contaminations might be because of disarray as morphologically comparable material recognized by vernacular names in various dialects enters the chain of commercialization, absence of valid crude plant material, and furthermore defilement for monetary benefit [41–43]. Deoxyribonucleic acid (DNA) barcoding has been effectively utilized in recognizing and evaluating defilement in crude grown exchange of an assortment of therapeutic plants [42]. G. cambogia items have high demands because of its ability to reduce weight, in spite of the fact that they can be tainted with comparative yet lower-valued species (G. indica). The detection and quantification of defilement in these dietary enhancements can be done by spectrometric and chromatographic techniques [31,44,45]. Rigorous legislation should be laid before and after marketing of these supplements in the market to ameliorate their safety and efficacy.

12.6. Future perspectives

G. cambogia fruit has many ingredients like HCA and benzophenones (guttiferones, garcinol, and xanthones), which are commercially very useful. Among these, HCA is considered as the active ingredient. Because of these ingredients, G. cambogia possesses various biological activities such as antiobesity, appetite-suppressant, and hyperlipidemic activity [1]. Over the last few years, many studies have been conducted to understand the potential of this fruit in promoting health. Numerous metabolic functions have been documented related to HCA like reducing food intake, appetite suppressant, increased fat oxidation, prevention of lipogenesis, thereby reducing the blood lipids [2]. Certain evidence-based studies report the antiobesity properties of G. cambogia and related toxicities and further observations which indicate the safe usage of the same. Numerous studies designed in a controlled manner prove the safety and efficacy HCA, with animals as well as human. However, there is a slight amount of adverse effect pertaining to gastrointestinal events. In addition, G. cambogia is suspected to increase the serotonin levels when combined with other drugs; therefore, it is necessary to investigate the interaction of other medicines with G. cambogia that elevates selective serotonin reuptake inhibitors serotonin levels. Nevertheless, evidence-based studies prove that HCA-SX alone does not produce any side effects and is regarded as a no-observed-adverse-effect level with a dietary dose of 2800mg/day of HCA-SX in humans [46]. A random metaanalysis of G. cambogia extract as well as HCA is investigated as substantial in reducing the body weight for a short period of time. Still more studies are to be conducted to support the long-term benefits of this extract along with HCA [4]. Besides, there are other phytochemical properties such as the antimicrobial, anticancer antioxidant, antidiabetic, hepatoprotective, antiulcerogenic, antiinflammatory activities, etc., that need to be studied experimentally as well as comprehensively [1]. Furthermore, regulatory bodies should adopt more stringent legislation for natural supplements at premarketing and postmarketing stages for identifying and scrutinizing the common adulterants and to render them safely for human use [2].

12.7. Declaration of interest

The authors profess no declaration of interest in this present chapter.

Acknowledgment

The authors of this article take this chance as an esteemed privilege to show their gratitude to the management of Aurea Biolabs (P) Ltd., Cochin, India, who were an invariable source of support to put pen to paper and encouraged us during the entire course of this review article. We also grab this golden opportunity to express deep appreciation to our colleagues for their valuable support that accelerated the successful completion of this task.

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Chapter 13: Camellia sinensis

C.V. Umesh
Department of Chemistry, MES Ponnani College, Malappuram, Kerala, India

Abstract

Camellia sinensis (tea) is used as a common drink in many of the houses with different flavors. Tea when taken in right forms can provide health benefits to an individual in many ways. The wide range of phytochemicals present in the tea such as polyphenols, flavonoids, etc., contribute to the array of pharmacological activities. The diverse range of biological activities of green tea polyphenols makes it a core ingredient in nutraceutical, functional food, and beverage applications.

Keyword

Anticancer; Antioxidant; Pharmacokinetics; Phytochemicals; Tea

13.1. Introduction

Camellia sinensis (tea) belonging to the Theaceae family is a plant that generally cultivates in tropical and subtropical climates. In the contemporary world, different tea is prepared from the leaves of Camellia sinensis considered as a very popular nonalcoholic drink next to water [1]. In the last 5000 years, sinking tea leaves into a hot water has provided luscious beverage [2]. According to the degree of fermentation, they are categorized as green tea (unfermented tea), white tea and yellow tea (lightly fermented), oolong tea (semifermented tea), black tea (fermented tea), and pu-erh tea (postfermented tea). Reports have mentioned, out of the global consumption of tea, the consumption of black tea predominates, followed by green tea [3]. Pu-erh tea is manufactured mainly in the Yunnan district in the southwestern part of China. Besides, it is popular in Taiwan also. The chemical constituents in the tea leaves are polyphenols (catechins and flavonoids), alkaloids (caffeine, theobromine, theophylline, etc.), volatile oils, polysaccharides, amino acids, lipids, vitamins (e.g., vitamin C), inorganic elements (e.g., aluminum and manganese), etc. [4]. Several studies revealed notable pharmacological activities of the Camellia sinensis such as antioxidant [5], antiviral [6], antibacterial [7], antiinflammatory [8], antimutagenic [9], antistress [10], antihypertensive [11], hypocholesterolemic [12], anticarcinogenic [13], antiaging [14], cardiovascular [15], anticancer [16], and anticoccidial activities [17]. They also showed photoprotective, anticellulite, slimming, improving skin and hair activities, and thereby opening the umbrella of green tea extracts to be used in cosmetic industries [18]. The chapter tries to present an overview of the phytochemical constituents, pharmacological activities, and clinical studies of various extracts of Camellia sinensis.

13.2. Phytochemicals

The phytochemical evaluation of Camellia sinensis has either quantified or qualified the presence of various phytochemicals such as flavonoids, phenolic acids, methyl xanthine (caffeine), amino acids, carbohydrates, lipids, proteins, beta-carotene, volatile compounds, vitamins, etc. (Fig. 13.1.) [19]. It is reported that tea contains quite a lot of bioactive compounds, while polyphenols contribute to about 33% and catechins are the mostly present polyphenols. 12 catechins have been identified, even though only eight catechins are present in tea in significant quantities. Most of the published studies focus precisely on epigallocatechin gallate (EGCG) and epigallocatechin (EGC) that constitute 70% of the total catechin amount in tea. Other important constituents are epicatechin (EC), epicatechin gallate (ECG), two flavonols such as quercetin-3-D-galactoside and kaempferol-3-glucoside, the oxyaromatic acids such as o-caffeoylquinic acid, neochlorogenic acid, and gallic acid (GA), two alkaloids such as caffeine and theophylline, and the amino-acid, L-theanine [20,21].

Figure 13.1
Important bioactive components of Camellia sinensis.

13.3. Pharmacokinetics

Even though the majority of the researchers have focused on the promising pharmacological activities of the different bioactives present in green tea, less has been studied on the bioavailability of actives. Studies have suggested that, less fraction of tea catechins is absorbed and hence to be considered bioavailable [22]. A study demonstrates that the concentration of catechins in green tea infusions is about 3250–4410mg/L [23]. A preclinical study has shown that only less than 5% of the tea catechins reached systemic circulation in rat, when administered orally, and approximately 1.68% of the ingested catechins were present in human plasma, which was distributed in human plasma (0.16%), urine (1.1%), and feces (0.42%) [24]. A pharmacokinetics study in rats showed that 14% of EGC, 31% of EC, and <1% of (−)-epigallocatechin-3-gallate appeared in the blood [25]. A human study with 3g of decaffeinated green tea, the maximum plasma concentration of EGCG, EGC, and EC were 0.57, 1.60, and 0.6μM, respectively [26].

Metabolic studies reveal that tea catechins undergo extensive metabolism by Phase II metabolism, such as UDP-glucuronosyltransferases, sulfotransferases, and catechol-O-methyltransferase before and after being absorbed mainly in the intestines and liver [27]. The metabolites identified in humans include glucuronide and sulfate conjugates, methylated tea catechin conjugates, and microflora-mediated ring fission products and phenolic acid catabolites [28]. One study also proved that when subjects supplemented with a green tea solution containing 634μmol of catechins, it was observed that 70% of the catechins are detected in the ileal fluid during 0–24h after administration, in which 33% was in the parent form and almost 40% as metabolites. Sixteen of the metabolites found in plasma was found in the forms of principally methylated, sulfated, and glucuronide conjugates of catechins, with peak catechins concentration in the range of 101–256ng/mL in plasma [29]. The mechanism for catechins being transported across epithelium is principally based on passive diffusion including paracellular and transcellular diffusions [30].

13.4. Pharmacological activities

The wide array of phytochemicals present in the leaves of Camellia sinensis contributes to the diverse range of pharmacological activities of the herb. Preclinical and clinical studies have focused on the diverse activities, which popularized the herb to be used in functional foods and nutraceutical supplements.

13.4.1. Antioxidant activity

Tea leaves are popular for its antioxidant properties. Green tea is best among them, and it acts as a potent source of valuable antioxidants, like fruits and vegetables. Some compounds in tea leaves are accountable for these properties. Among phytochemicals, polyphenols make the most stimulating group of tea leaf constituents and shows effective antioxidant activity [31]. Catechins and their derivatives EGC, EGCG, ECG, (−)-gallocatechin gallate (GCG), and EC are the chief polyphenolic contents in tea [32,33].

About 59% of total catechins constituted by EGCG, 19% EGC, 13.6% ECG and 6.4% EC. Among these catechins, EGCG and ECG are categorized as ester catechins and EGC and EC are nonester catechins [34]. Their oxidation resistance has been arranged as EGCG≈ECG>EGC> GA>EC>catechin [3,35]. The antioxidant activity of these compounds is correlated to the molecular structure. The number and position of hydroxyl groups in these polyphenolic compounds generally regulate their antioxidant properties. Besides these catechins, some polyphenols in tea, such as quercetin, kaempferol, and myricetin also reveal oxidation resistance [36,37]. The antioxidant effect of these tea polyphenols comprises the combined effect of the following processes: increase in the activity of antioxidant enzymes, they inhibit lipid peroxidation, scavenging of free radicals in synergy with other nutrients and the reduction of oxidation via chelation of metal ions [13,38]. During oxidation or fermentation, catechins changed to theaflavin, theaflavin-3-gallate, theaflavin-3′-gallate, theaflavin-3-3′-digallate and thearubigins. These are liable for the bitter taste and dark color of black tea [39]. The amount of catechins present in the leaves have been found to be higher in the leaves used for black tea production than in those used for green tea production [40]. Bioaccessibility and absorption of antioxidants of green tea catechins in animal and human beings are fairly well acknowledged.

13.4.2. Anticancer activity

Several studies have revealed that green tea assists to protect against cancer. Green tea drinking displayed chemopreventive stuffs on various cancers. Drinking green tea is connected with decreased rate of cancer development. Evolving researches suggest that green tea polyphenols show significant role in the prevention of cancer. It also presumes that polyphenols help to kill cancerous cells and stop their progression [41–43]. Catechins extracted from green tea plants have biologic activity in antioxidant, antiangiogenesis, and antiproliferative studies that are potentially applicable to the prevention and treatment of different types of cancer [44]. EGCG, a major polyphenol found in green tea, and it is a broadly studied chemopreventive agent with potential anticancer activity [45].

Epidemiological and laboratory investigations have isolated EGCG in green tea polyphenols as the most potent chemopreventive agent that can induce apoptosis and suppress the formation and growth of human cancers including colorectal cancers [46]. Different types of mechanisms have outlined regarding the inhibition of cancer cell survival by EGCG and other green and black tea polyphenols. EGCG blocked androgen receptor expression and several growth factor receptors. Researchers have been found that telomerase activity inhibited by EGCG led to telomere fragmentation [47].

EGCG can regulate the expressions of vascular endothelial growth factor, matrix metalloproteinases, uPA, IGF-1, EGFR, cell cycle regulatory proteins, and thereby inhibits NF-κB, PI3-K/Akt, Ras/Raf/MAPK, and AP-1 signaling and showing strong cancer chemopreventive effects [48]. Routine green tea drinking may cause the inhibition of CYP1A activity by polyphenols, which has been recommended as a chemoprevention against carcinogens that were bioactivated by CYP1As. Therefore, any changes in hepatic CYP1As may be considered as a biomarker for green tea chemoprevention [49]. Certain studies have identified that green tea slowed the progression of ductal lesions to progressive mammary intraepithelial neoplasias and suppressed tumor invasiveness. It can also inhibit the proliferation of ductal epithelial cells and tumors [50]. EGCG has antioxidant and also ultraviolet (UV)-B signal transduction blocking activity as well as sunscreen activity. Nonmelanoma is a very common type of skin cancer, and it is mostly associated with chronic exposure to UV light. Oral or local application of green tea extracts inhibits skin tumor formation induced by chemical carcinogens or UV radiation, thereby reducing the risk of nonmelanoma cancer [51,52]. In this regard, it stimulated many cosmetic and pharmaceutical companies to supplement their skin care products with green tea extracts [53]. Green tea polyphenols act at several points regulating cancer cell growth, persistence, and metastasis, including effects at the ribonucleic acid (RNA) and deoxyribonucleic acid (DNA), and protein levels. Population-based studies suggest that green tea consumption is associated with reduced risk of several human malignancies such as cancer and diabetes [54].

13.4.3. Antibacterial activity

Several studies identified that green tea leaves are known for antibacterial activity against various human pathogenic bacteria. Green tea polyphenols show the significant role in green tea's antibacterial activity [55]. The polyphenols have the ability to damage bacterial cell membrane. The green tea has been showed higher antimicrobial activity than the black tea [56]. Green tea is active against multidrug-resistant Escherichia coli and Staphylococcus aureus [57]. Green tea and a purified compound EGC have revealed worthy inhibitory and bactericidal effect on the multidrug-resistant pathogens. Green tea extract has shown the better bactericidal effect on gram-positive pathogen than gram-negative pathogen, which may be due to their dissimilar cell wall components. EGCG exhibited greater antibacterial activity against Staphylococcus aureus compared to Escherichia coli [58–60]. The mixture of purified EGCG compound from green tea and antibiotic gentamicin has strong synergistic relationship and is highly active against Escherichia coli and Staphylococcus aureus [61]. EGCG stimulates destruction of the cell wall, which intensifies the permeability of cell membrane to antibiotics that influence the additive effects [62]. Urinary tract infection is a most common bacterial infection in women in reproductive age affecting illness in females commonly in the developing countries due to illiteracy, unhygienic conditions and lack of proper toilet facilities [63,64]. Green tea extracts inhibit the growth of uropathogenic Escherichia coli by its polyphenolic catechins. Antioxidant properties of catechins cause the elimination of free radicals and also they can break up the cell wall membrane that suppressed the bacterial growth [65,66]. When patients with severe infections due to extended spectrum β-lactamase (ESBL) producing bacteria are resistant to all common antibiotics, resistance to antimicrobial agents has become a gradually relevant problem for healthcare takers. Researchers found that aqueous green tea extracts have the ability to inhibit such pathogens. Antibiotic therapy combined with such phytochemicals from aqueous green tea extracts may interfere the emergency of bacterial resistance and may also produce desirable synergistic effects in the treatment of infections caused by ESBL [67–69].

13.4.4. Antiviral activity

Virus infectious diseases still continued as a major public health concern with the chance of epidemics and pandemics. Viral infections might be prevented or controlled in human involving vaccination, antiviral treatment and personal hygiene. Vaccination is very effective for stopping and controlling viral infections; personal hygiene provides many cost-effective alternatives to preventing such infections. Green tea polyphenols have a range of activities against RNA and DNA viruses [70]. The antiviral property of polyphenols is due to their ability to inhibit the enzymes entailed in viral replication and to their cell membrane disruption. Adenoviruses are involved in a variety of human and animal pathologies [71]. Green tea extract exposes the dominant inhibitory effect on viral persistence hepatitis B antigen [72]. It was reported that EGCG inhibits hepatitis C virus entry into target cells and prevents cell-to-cell spread between adjacent cells [73]. It has been revealed that the intake of catechins for five months induce inhibitory effects on influenza virus infection [74–76].

EGCG inhibits Bovine coronavirus (BCV), the causal pathogen for diarrhea in cattle more efficiently in the bovine intestinal tract, where the temperature of approximately 37°C is appropriate for the antiviral efficacy of EGCG against BCV [77]. Recent studies show that the main constituents of green tea, catechin, and EGCG inhibit chikungunya virus infection [78].

13.4.5. Antiobesity activity

Studies prove that individuals suffering from weight management disorders are increasing day by day, with 25% of individuals suffering from overweight and 10% with obesity [79]. Studies have highlighted the role of green tea extract and its role in weight management. In one study, the green tea extract stimulated brown adipose tissue thermogenesis to an extent which was much greater that could be attributed to its caffeine content, and it was assumed that was predominantly due to the catechins and polyphenols present in the green tea [80]. The antiobesity effects of green tea in mice indicated that the body weight increase and increase in weight of intraperitoneal adipose tissue were significantly reduced, and it can be possible due to the synergistic effects of catechins and caffeine [81]. In a pilot study, the effects of green tea extract with EGCG, on thermogenesis and fat oxidation in obese men were evaluated and found that EGCG alone has the potential to increase fat oxidation in men and thereby contributing to the antiobesity effects [82]. Similarly, the effects of a thermogenic beverage containing green tea catechins, caffeine, and calcium proved that the formulation increases 24h energy expenditure by 46%, but the study could not prove the contribution of individual components [83]. The effects of green tea extract in reducing body fat and thereby reducing the risks of cardiovascular diseases were evaluated and found that green tea extract enriched with catechins has significantly reduced the body weight, systolic blood pressure, and low-density lipoprotein cholesterol, thereby indicating the promising role of the extract in fat reduction and reducing the risks of green tea extract [84]. Similarly, the effects of green tea were investigated on high fat–induced obesity, symptoms of the metabolic syndrome, and fatty liver in mice and found that long-term supplementation of green tea EGCG treatment attenuated the development of obesity, symptoms associated with the metabolic syndrome, and fatty liver [85].

13.4.6. Antimicrobial activity

The widespread use of synthetic antibiotics has led to an increase in the number of resistant pathogenic bacteria. Diseases/infections arised as a result of gram-positive and gram-negative pathogens have become a serious threat worldwide [86]. The antimicrobial activity of green tea was first reported in the laboratory by McNaugh in early 19th century by a British Army man, where he showed that brewed black tea killed the Salmonella typhi and Brucella melitensis and he suggested that the bottles of the army men can be filled with black tea instead of water to fight against the infectious diseases which can be arised during the war [87]. The antimicrobial and antifungal activity of various fractions of green tea on vaginal pathogens such as Proteus mirabilis, Streptococcus pyogenes and Candida albicans were determined and found that the ethyl acetate fractions were found to be more effective than the aqueous fractions and among the catechins, EGCG showed the promising activity [88]. An in vitro study on Chinese tea (Lung Chen tea), two tea catechins, EGCG and EC were determined on Helicobacter pylori and found that EGCG can be one of the promising bioactive ingredients in inhibiting the growth of pathogen [89]. Similarly, susceptibility of green tea bioactives like catechins against Candida albicans were determined under various pH conditions and the synergism of the combination of catechin and antimycotics. The study found that EGCG enhances the antifungal effects of amphotericin B or fluconazole against antimycotic-susceptible and resistant Candida albicans [90]. The antifolate activity of green tea extract was evaluated against Stenotrophomonas maltophilia and showed that EGCG is an efficient inhibitor of Stenotrophomonas maltophilia dihydrofolate reductase, a strategic enzyme which is considered an attractive target for the development of antibacterial agent [91]. Tea bioactives were also found to be a promising candidate for the inhibition of various pathogens such as Staphylococcus aureus [92], Clostridium perfringens [93], Pseudomonas aeruginosa [94], etc.

13.4.7. Antihypertensive and cardiovascular activity

Studies have also evaluated the role of Camellia sinensis and its active component's role in antihypertensive and cardiovascular activity. The role of green and black tea in lowering of blood pressure in stroke-prone spontaneously hypertensive rats was determined and found that on account of the potential antioxidant activity, green tea extract attenuated the increase of blood pressure [95]. In another study, population-based prospective cohort study (Ohsaki Study) was conducted to examine the association between green tea usage and mortality from cardiovascular disease in northern Japan. The study gave a promising inverse relationship between the consumption of green tea and cardiovascular diseases [96]. Preclinical study also proves that green tea consumption improves cardiovascular and metabolic function in spontaneously hypertensive rats. The study found that extract increased nitric oxide production and thereby contributing to the improved cardiovascular activities [97].

13.4.8. Other promising activities

The promising health benefits of green tea also give an eye opening for exploring its pharmacological activities and thereby delivering a value-added product. The neuroprotective effects of green tea were studied in Parkinson's and Alzheimer's disease. The study found that the extract enhanced the release of the nonamyloidogenic soluble form of the amyloid precursor protein (sAPPa) and thereby promoting promising neuroprotective effects [98]. Studies also suggest that tea bioactives play a promising role in reducing dental diseases such as dental cancer [99] and periodontal disease [99]. EGCG is also found to suppress experimental autoimmune encephalomyelitis induced by proteolipid protein [100].

13.5. Conclusion

The miraculous benefits of green tea and its bioactive components are well-cited literature on account of their miraculous health benefits. The bioactive polyphenols and catechins have paved the eye opening for the use of green tea in food, beverage and nutraceutical applications. Several clinical studies have also focused on the use of various forms of green tea in the nutraceutical and natural product industry. Furthermore, clinical studies with significant results can give a value-added product from green tea, which can be explored for the various health benefits.

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Chapter 14: Ferula asafoetida

Ayman M. Mahmoud
[1]
,
[2]
, Reem S. Alruhaimi
[3]
, and Emad H.M. Hassanein
[4]

1
Department of Life Sciences, Faculty of Science and Engineering, Manchester Metropolitan University, Manchester, United Kingdom
2
Physiology Division, Zoology Department, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt
3
Department of Biology, College of Science, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia
4
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University-Assiut Branch, Assiut, Egypt

Abstract

Ferula asafoetida is a herbaceous plant that belongs to the Ferula genus of the Umbelliferae family. Plants of this genus are rich sources of natural products possessing multiple biological and pharmacological activities. The oleo-gum-resin asafoetida obtained from the rhizome and root of F. asafoetida is traditionally used as a digestive aid and in treating several diseases, including bronchitis, asthma, neurological disorders, and others. Besides its traditional uses, experiments have shown the antispasmodic, laxative, antioxidant, relaxant, hepatoprotective, neuroprotective, anticancer, and other pharmacological activities of asafoetida. Ferulic acid, coumarins, sesquiterpene coumarins, monosaccharides, polysaccharides, glycoproteins, monoterpenes, and other compounds are the main constituents of asafoetida. The present chapter presents an overview of asafoetida and its pharmacological activities.

Keywords

Ferula asafoetida; Medicinal plants; Oleoresin; Umbelliferae

14.1. Introduction

Asafoetida is the dried gum oleoresin exuded from the tap root or rhizome of many species of the genus Ferula and used as a flavoring agent in several countries. Ferula spices are present widely in Western Afghanistan, Eastern Iran, Turkey, and Iraq, as well as North African countries and Europe [1]. Ferula asafoetida is a species of Ferula endemic to Afghanistan and Iran. This species grows to about two meters and comes in two varieties: bitter and sweet [2]. Ferula plants have massive taproots or carrot-shaped roots around 15cm in diameter at the crown when they are 4–5 years old, and asafoetida is extracted from them. The upper part of the living rhizome root is fully exposed before the plants flower, and the stem is cut off close to the crown. A dome-shaped structure made of twigs and earth covers the exposed surface, and the cut surface exudes a milky juice. When more latex exudes, the exudates are scraped off and a new slice of the root is cut; sometimes, the resin is removed along with the slice. The procedure of collecting resin and slicing the root is repeated until exudation stops [3]. The odor of asafoetida is pungent, persistent, and sulfurous, and it is traditionally used for the treatment of many diseases [4–7]. The water extract of asafoetida is used traditionally in Afghanistan for the treatment of hysteria, ulcers, and whooping cough, and in Egypt as diuretic, antispasmodic, vermifuge, and analgesic [8]. In Nepal, asafoetida is believed to have diuretic, sedative, and aphrodisiac properties and hence is consumed in the daily diets [9]. Asthma, flatulence, influenza, epilepsy, and stomachache are among the diseases traditionally treated with asafoetida [10]. Studies have demonstrated its antioxidant, antiviral, anticancer, antifungal, antidiabetic, hypotensive, antispasmodic, and other beneficial effects [10].

14.2. Chemical constituents

Carbohydrates, moisture, proteins, fats, minerals, and fibers are the main constituents of asafoetida. Resin, gum, and essential oil are the three main fractions [4] (Table 14.1). Ferulic acid and its esters, terpenoids, coumarins, and sesquiterpene coumarins are found in the resin fraction. The gum contains galactose, glucose, l-arabinose, glucuronic acid, rhamnose, glycoproteins, and polysaccharides, while monoterpenes, sulfur-containing compounds, and other volatile terpenoids are found in the volatile fraction [10] (Table 14.2). Sulfur compounds found in F. asafoetida resin have many pharmacological properties and are potentially valuable for medicine [4,11].

Table 14.1

Main constituents of Ferula asafoetida.

Constituents | Percentage
Carbohydrates | 68%
Moisture | 16%
Protein | 4%
Fats | 1%
Minerals | 7%
Fibers | 4%

Table 14.2

Main fractions of Ferula asafoetida.

Fraction | Percentage | Compounds
Resin | 40%–64% | Ferulic acid and its esters, coumarins, sesquiterpene coumarins, and other terpenoids
Gum | 25% | Glucose, galactose, 1-arabinose, rhamnose, glucuronic acid, polysaccharides, and glycoproteins
Essential oil | 10%–17% | Sulfur-containing compounds, monoterpenes, and other volatile terpenoids

14.3. Pharmacological and therapeutic potentials of asafoetida

The biological and pharmacological activities of asafoetida have been demonstrated in various studies. Asafoetida exerted promising therapeutic potential in a number of diseases.

14.3.1. Anticancer activity

The beneficial effects of asafoetida in experimental models of tumors, including skin and colon cancers have been reported. Unnikrishnan and Kuttan have investigated the antitumor activity of asafoetida extract in mice with Ehrlich ascites tumors [12]. The extract of asafoetida inhibited both stages of chemical carcinogenesis induced by croton oil and 7,12-dimethyl benzanthracene, resulting in significant reduction in the formation of papilloma. The findings of this study suggested that asafoetida could be used as an anticancer agent. Accordingly, asafoetida restored antioxidant levels and significantly reduced ornithine decarboxylase activity and DNA synthesis, pinpointing its antioxidant and anticarcinogenic potentials [13]. The effects of F. asafoetida on N-methyl-N-nitrosourea-induced mammary epithelial tissue differentiation in rats were investigated by Mallikarjuna et al. [14]. In this study, treatment with asafetida reduced tumor multiplicity, decreased the number of terminal end buds during differentiation, and lowered the number of tumors per tumor-bearing rat [14].

Panwar and coworkers have studied the chemopreventive potential of various doses of the oleo-gum-resin of F. asafoetida on colon cancer induced by 1,2-dimethylhydrazine in rodents [15]. Asafoetida potently attenuated the deleterious effects of 1,2-dimethylhydrazine and the development of colon carcinogenesis in rats. Furthermore, the antitumor activity of F. asafoetida oleo-gum-resin against breast cancer in BALB/c mice induced by 4T1 cells was examined by Bagheri et al. [16]. The supplementation of asafoetida resulted in potent antitumor and antimetastasis effects manifested by the reduced tumor weight and volume, provoked necrosis in tumor tissue, improved body weight, and prevented lung, liver, and kidney metastasis [16]. In an in vitro study, the antitumor activity of asafoetida and its essential oil, as well as ferulic acid, was investigated using murine breast cancer cell line [17]. Asafoetida and its essential oil stopped the proliferation of 4T1 cells. MTT assay revealed a significant cytotoxicity activity that was time- and concentration-dependent [17]. In line with these studies, the antiproliferative activity of microwave-extracted essential oil of F. asafoetida was investigated using two liver cancer cell lines (HepG2 and SK-Hep1) [18]. Dithiolane, which is abundant in F. asafoetida, showed antiproliferative activity in human liver carcinoma cell lines in a dose-dependent manner and its effect was mediated through the induction of apoptosis [18].

14.3.2. Neuroprotective effects

14.3.2.1. Effect on glutamate-induced neuronal cell damage

Traditional uses and experimental findings pinpointed that F. asafoetida could be used as a neuroprotective agent. Tayeboon et al. tested the neuroprotective effects of F. asafoetida extract on glutamate-induced neuronal cell damage and reported its ability to attenuate glutamate-induced apoptosis/necrosis [19].

14.3.2.2. Effect on peripheral neuropathy

Moghadam et al. have conducted in vitro studies to determine the response pattern of isolated sciatic nerves to different concentrations of asafoetida in Lock's solution [20]. In vivo tests were also carried out on mice to see if it helped them recover from peripheral neuropathy. The in vitro studies demonstrated that aqueous extract of oleo-gum-resin increased the amplitude of nerve action potential and decreased the latent period. In asafoetida-treated animals, nerve conduction velocity and action potential amplitude were improved. The in vivo studies revealed that asafoetida is a nerve stimulant, and its administration to neuropathic rodents resulted in neuroprotection by stimulating axonal remyelination and regeneration, as well as a reduction in lymphocyte infiltration [20].

14.3.2.3. Memory-enhancing activity

The effects of F. asafetida on learning and memory in rats were studied by Vijayalakshmi et al. [21]. In an elevated plus-maze model, the extract significantly improved memory score and transfer latency dose-dependently, inhibited brain cholinesterase, and restored the antioxidant balance. In another study, Bagheri et al. [22] investigated the preventive effect of asafoetida on D-galactose and NaNO_2-induced dementia in a murine model and concluded that it can prevent and treat amnesia. The beneficial effects of asafoetida were attributed to the presence of some active constituents such as ferulic acid and umbelliferone [22].

14.3.2.4. Antinociceptive and anti-inflammatory effects

The antinociceptive and anti-inflammatory effects of asafetida have been demonstrated using the hot plate test. Asafetida had a significant antinociceptive effect in a hot plate test at all tested doses, with 10mg/kg being the most effective. Fifteen minutes after asafetida administration, the maximum potent effect was observed. None of the antagonists could reverse asafetida's antinociceptive effect, but it has shown a remarkable antioxidant and inhibitory effect on lipoxygenase activity [23].

14.3.2.5. Anticonvulsant effects

In pentylenetetrazole (PTZ) model, the antiepilepsy and antioxidant effects of the gum extract were investigated by Kiasalari et al. [24]. F. asafoetida gum extract restored oxidant/antioxidant balance effectively. The antioxidant activity of hydroalcoholic F. asafoetida gum extract and the reduction of oxidative stress are likely to be responsible for their effects on PTZ-induced seizures [24]. Bagheri et al. [25] investigated the anticonvulsant efficacy of asafoetida on chemical and electrical kindling models. The mean seizure stage during the injection of PTZ was significantly reduced when animals were pretreated with asafoetida. In the amygdala kindle model, seizure parameters were improved with 50 and 100mg/kg doses. In asafoetida-treated rats, the number of stimulations in stages 3, 4, and 5 increased significantly at both doses [25].

14.3.2.6. Anxiolytic effect

The anxiolytic, analgesic, and sedative properties of asafoetida were investigated by Algasoumi [26] who used hot-plate, hole-board, and elevated plus-maze tests, and motor activity meter in rodents. The findings revealed that asafoetida has anxiolytic and analgesic activity that is dose-dependent, with a calm sedative potential at high doses. When compared to diazepam, asafoetida appeared to be an effective alternative for anxiety disorders treatment. Asafoetida in low doses can be a therapeutic alternative to the currently used anxiolytic drugs.

14.3.3. Effect on the gastrointestinal tract

In general, spices have long been thought to improve salivary flow and the secretion of gastric juice, as well as aid in digestion. The digestive stimulating effect of spices is most likely due to an increase in the activities of enzymatic participants in digestion. The beneficial effect exerted by a large number of spices consumed in the diet on the pancreatic digestive enzymes represents a key factor contributing to the well-known digestive stimulant action. Interestingly, the pancreatic lipase activity was significantly increased, and pancreatic amylase was stimulated by asafoetida [27]. In the same context, Ramakrishna Rao et al. [28] investigated the in vitro effects of asafoetida on the activity of the digestive enzymes. When spices come into direct contact with pancreatic lipase and amylase, they significantly increase their activity. This positive influence on enzyme activity, in addition to causing an increase in the titers of pancreatic digestive enzymes, is thought to play a supporting role in the digestive stimulant effect [28].

14.3.4. Relaxant and antispasmodic effects

Growing evidence supported the relaxant effects of asafoetida. In 2012, the relaxant effects of F. asafoetida were studied using different smooth muscles [29]. The relaxant effect of the aqueous extract of asafoetida on guinea pig tracheal smooth muscle was studied, and the results showed that it has a potent relaxant effect, possibly due to muscarinic receptor blockade [29]. The relaxant effects of asafoetida oleo-gum-resin and its coumarin constituent umbelliprenin on guinea pig tracheal chains were studied by Bayrami et al. [30]. The aqueous extract and umbelliprenin exerted concentration-dependent relaxant effects on precontracted muscles. Also, in tracheal smooth muscle samples, Khazdair et al. [31] studied the relaxant effect of asafetida on muscarinic (M) receptors. The effects of aqueous extract of F. asafoetida as well as atropine on M-receptors were tested. The maximum response was concentration-dependent. Because F. asafoetida or its constituents can bind to M-receptors and prevent methacholine from binding to these receptors, it was suggested that F. asafoetida has a competitive antagonistic effect at M-receptors [31].

Previous studies have supported that an antispasmodic therapy could be achieved with asafetida on the ileum and concluded that asafoetida could be used as an antispasmodic agent. Bagheri et al. [32] have investigated the relaxant effect of asafoetida and its essential seed oil in the isolated rat ileum. On acetylcholine (Ach)-induced contractions, asafoetida had an antispasmodic effect and potently reduced Ach-induced contraction [32]. Another study was done by Fatehi et al. [33] reported that the spontaneous contractions induced by histamine, Ach, and potassium chloride in the isolated guinea pig ileum were reduced. F. asafoetida gum extract exerted a dose-dependent relaxant effect and decreased mean arterial blood pressure [33].

14.3.5. Hepatoprotective effect

Dandagi et al. [34] investigated the hepatoprotective properties of an F. asafoetida extract against carbon tetrachloride–induced hepatotoxicity. F. asafoetida extracts attenuated liver injury and ameliorated serum transaminases. In a rat model of ethylene glycol-induced lithiasis, Bagheri et al. [35] have reported the hepatoprotective efficacy of F. asafoetida as shown by the amelioration in circulating markers of liver function.

14.3.6. Antimicrobial activity

The essential oil produced during the early stages of F. asafoetida growth could be used in the food industry, while the essential oil produced during the later stages could be used as antibacterial agent [36]. Asafoetida can be used to treat a variety of fungal and bacterial infections. Using the agar disc diffusion method, both alcoholic and aqueous extracts inhibited the growth of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Aspergillus niger [37]. By inhibiting the respective fungi and bacteria, the crude extract demonstrated a broad spectrum of antimicrobial activities [37]. In addition, the antimicrobial properties of the volatile oils isolated by hydrodistillation from two varieties of F. asafoetida, Pathani and Irani, were investigated against a variety of food-borne bacterial and fungal organisms, including E. coli and Bacillus subtilis, Pathani was more effective [38]. The volatile oil of Irani inhibited the growth of Aspergillus ochraceus and Penicillium chrysogenum more potently than Pathani [38]. Moreover, Bhatnager et al. [39] have tested the antimicrobial activity of two types of F. asafoetida, red and white gum, against five different bacterial strains. The highest antibacterial activity was found in hexane extracts of both red and white asafoetida against Shigella flexneri, while S. aureus was least affected by the other extracts. Patil et al. [40] investigated the antibacterial activity of asafoetida extracts against B. subtilis, E. coli, Klebsiella pneumoniae, and
S. aureus. Owing to the presence of a mixture of phytoconstituents, ethyl acetate, ethanol, and methanol extracts had significant antimicrobial activity than other extracts and could be used to develop new antibiotic compounds [40]. Moreover, the antiquorum sensing activity of F. asafoetida against P. aeruginosa was investigated and the results showed a potent antiquorum activity, along with stopping Chromobacterium violaceum from producing violacein. F. asafoetida oil treatments reduced pyocyanin, pyoverdine, elastase, and biofilm production [41].

Pertaining to the antifungal activity, Sitara et al. [42] tested essential oils of asafoetida and others for antifungal activity against different seed-borne fungi (Aspergillus flavus, A. niger, Fusarium moniliforme, Fusarium oxysporum, and Fusarium niv), and the results revealed a potent fungicidal activity [42]. Additionally, the antifungal and allelopathic effects of asafoetida were tested against Pleurotus spp. and Trichoderma harzianum [43]. At higher concentrations, asafoetida exhibited fungistatic and fungicidal properties against T. harzianum and Pleurotus spp. [43]. Furthermore, the activity of F. asafetida against Sclerotium rolfsii ITCC 5226 and Macrophomina phaseolina ITCC 0482 was screened in vitro [44]. Formulations containing F. asafoetida effectively controlled pathogenic fungi [44]. In complete randomized design and an in vitro method [45], the antifungal effects of asafoetida seed essential oil on some plant pathogen fungi were studied and the results showed potent antifungal effects against all tested species.

14.3.7. Antiparasitic activity

The activity of asafoetida against Blastocystis sp. in vitro growth was investigated by El Deeb et al. [46]. In this study, the oil and powder forms of asafoetida reduced the count and viability of all tested isolates. The inhibitory effect of asafoetida extracts was highly correlated to the concentration, form, and time of incubation [46]. Gundamaraju et al. [47] tested the anthelmintic activity of F. asafetida extract against Pheretima posthuma, which included determining the time of worm paralysis and death. At the highest concentration, the extract showed significant antihelmintic activity exceeded that of the standard drug piperazine citrate [47]. Additionally, F. asafoetida exerted toxic effects on the liver fluke Fasciola gigantica [48].

14.3.8. Antidiabetic and anti-obesity effects

Asafetida showed a protective effect in diabetes-induced by alloxan where it preserved the integrity of pancreatic β-cells, supporting its traditional use to prevent diabetic complications [49]. In streptozotocin (STZ)-induced diabetic rats, Latifi et al. [50] investigated the hypoglycemic activity of asafoetida extract. At a dose of 50mg/kg for four weeks, asafoetida showed hypoglycemic activity which could be attributed to the presence of phenolic acid and tannins. In addition, the ethanolic extract of asafoetida protected against STZ-induced diabetes in rats where it effectively increased serum insulin levels, and improved liver and kidney function, effects that were associated with a potent antioxidant activity [50]. In type 2 diabetic rats, Azizian et al. [51] investigated the effects of F. asafoetida on weight gain, fat accumulation, liver steatosis, and leptin levels. F. asafoetida significantly reduced body weight, abnormal fat, and epididymal adipocyte size. Serum leptin levels in treated rats were significantly lower. The findings revealed the potent anti-obesity, lipid-lowering, and anti-steatosis properties of asafoetida. F. asafoetida gum is thus a promising treatment option for diabetes-related obesity and hepatic steatosis.

14.3.9. Effect on blood and blood vessels

Asafoetida possesses antihypertensive, anticoagulant, and fibrinolytic activities. The intravenous administration of the gland tincture in rabbits caused a significant smooth muscle relaxant effect and a decrease in blood pressure [52]. In dogs, intravenous administration of the gum-resin water extract induced vasodilatation, decreased blood pressure [53], and exerted anticoagulant activity [54]. Anesthetized rats that received the gum extract exhibited a significant reduction in the mean arterial blood pressure [33]. In addition to the experimental studies, fibrinolytic activity was observed in healthy human subjects who received the ether extracts of the dried gum and gum resin [55]. The effect of asafoetida on the cardiovascular parameters in acute hypertension induced by angiotensin II (AngII) has been recently demonstrated [56]. In AngII-induced rats, asafoetida (10, 30, and 60mg/kg) alleviated systolic blood pressure, mean arterial pressure, and heart rate [56].

14.3.10. Metabolic activity

In rat liver, Korashy et al. [57] investigated the effects of F. asafoetida on the expression of the cytochrome P450 2C11 (CYP2C11). The mRNA and protein expression levels of CYP2C11 were significantly downregulated by F. asafoetida in a dose-dependent manner.

14.3.11. Anti-aging effect

The anti-aging effect of asafetida on senescent human dermal fibroblasts was examined by Moghadam et al. [58]. In this study, asafetida showed rejuvenating effects on senescent fibroblasts, lowered their β-galactosidase activity, and increased the cell proliferation rate [58].

14.3.12. Reproductive effects

Asafoetida showed beneficial effects on the sperm parameters. Bagheri et al. [59] investigated the effects of asafoetida on sperm parameters, circulating testosterone, and testis tissue. Asafoetida enhanced the number and viability of sperm, increased the number of Leydig cells, and enhanced spermatogenesis in a dose-dependent manner [59].

14.4. Toxic effects

Oral administration of F. asafoetida extract to rats had no significant effect on body weight, body weight gain, or hematological parameters in rats, and there were no significant increases in the serum liver and kidney function biomarkers [60]. These findings revealed that F. asafoetida had broad safety and little toxicity for short-term use in a dose of 250mg/kg [60]. Another study reported that up to 72h after asafetida administration, no mortality was observed in an acute toxicity study [61]. Within 24h, there were no signs of neurological or behavioral changes. The intake of asafetida altered hematological parameters, and the aspartate aminotransferase and lactate dehydrogenase levels were significantly higher. Throughout the study, the administration of asafetida did not affect the plasma levels of urea and creatinine [61]. Although showed a good safety profile in experimental studies, a case of methemoglobinemia has been reported in a five-week-old black male infant after ingesting asafoetida. He recovered from the onset of tachypnea, grunting, and cyanosis with intravenous methylene blue treatment [62]. Asafetida is also not recommended for use during pregnancy because it may increase the risk of abortion [63].

14.5. Conclusion

The oleo-gum-resin of F. asafoetida, asafoetida, is used widely in several countries as a flavoring agent and traditionally for the treatment of various diseases. Asafoetida is rich in bioactive constituents, including ferulic acid, coumarins, terpenoids, sulfur-containing compounds, monoterpenes, and others. Owing to its rich content of these bioactive natural products, asafoetida possesses promising pharmacological activities, such as, antispasmodic, antioxidant, antihypertensive, relaxant, hepatoprotective, neuroprotective, and anticancer, and has been used in the treatment of bronchitis, asthma, neurological disorders, and other disorders. F. asafoetida can be used as a potential ingredient in various functional foods and nutraceuticals due to its various biological activities and functional properties. This great medicinal value of asafoetida is worthy of further investigations corroborated by clinical trials to evaluate the safety and exact mechanisms underlying its therapeutic effects.

14.6. Conflict of interest

None.

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Chapter 15: Concise review on Asparagus racemosus and its role as nutraceuticals and functional foods

Dinesh Chaudhary
[1]
, Suman Giri
[2]
, and Gopal Lamichhane
[3]

1
Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon, South Korea
2
College of Pharmacy, Daegu Catholic University, Gyeongsangbuk-do, South Korea
3
College of Pharmacy, Wonkwang University, Jeollabuk-do, South Korea

Abstract

Asparagus racemosus commonly known as Shatavari or Kurilo is a spinous shrub with tuberous roots found commonly in the tropical and subtropical regions of Nepal, India, Australia, and Africa. It is a well-known plant in Ayurveda, Siddha, Unani, and other traditional system of medicines for the treatment of various ailments, including dyspepsia, bleeding and nervous disorders, cough, jaundice, diabetes, to promote lactation, tonic, etc. Numerous bioactive phytochemicals mainly steroidal saponins and flavonoids have been isolated from different parts of this plant, which have a wide variety of pharmacological and therapeutic benefits. It is commonly cultivated and consumed by the local people as a nutritious vegetable to prepare alcohol and to promote milk secretion in cattle.

Keywords

Asparagus racemosus; Buttermilk Root; Functional Foods; Galactagogue; Milk secretion; Nutraceuticals; Phytoestrogens; Shatavarin

15.1. Introduction

Asparagus racemosus (Fig. 15.1), also known as Shatavari or Kurilo is a perennial shrub with a tuberous rootstock that has spine, branched woody stems and which grows up to 2m in height [1,2]. It is a common species of Asparagus which is found in the tropical and subtropical regions of Nepal, India, Sri Lanka, etc. Approximately, 300 species of Asparagus have been found around the world, among which about 4 and 22 species have been reported in Nepal and India, respectively [3,4]. Since prehistoric era, it has been used in treatment, prevention, and mitigation of diseases, which was firstly documented in Rigveda (4500 and 1600 BCE) for its use against several diseases [5,2]. Its bioactivity is mainly attributed to steroidal saponins and sapogenins. In 1799, it was botanically classified as follows:

Kingdom: Plantae
Clade: Angiosperms
Clade: Monocots
Order: Asparagales
Family: Asparagaceae
Subfamily: Asparagoideae
Genus: Asparagus
Species: Asparagus racemosus
Botanical name: Asparagus racemosus
Synonyms: Asparagus volubilis

Common name: Buttermilk Root (English), Kurilo (Nepalese), Shatavari (Hindi), Sadavare (Tamil), Tian Men Dong (Chinese), Satawar (Urdu), Shakakul/Shaqaqul (Arabic), Satamuli/Dongri (Bengali).

Figure 15.1
Matured Aparagus racemosus plants and its parts. Picture were taken from Er. Rajan Kumar Adhikari garden, Purunchaur, Kaski, Nepal. Photo were taken by Anup Giri.

15.2. Habit and habitat

It is found abundantly in the tropical and subtropical forests from altitude of 165 to 2200m throughout Asia, Australia, and Africa. It prefers to grow in light, medium, and heavy soils. However, continuous deforestation and pollution have made Asparagus to be listed in endangered from its natural habitat [6,5,7].

15.3. Macroscopic and microscopic characterization

15.3.1. Macroscopic characterization

It has adventitious root system. Tuberous roots are externally brown, rough with longitudinal wrinkles, while the inner surface is creamy white. It is heavily branched that are usually distinctly striate-rigged and are almost up to 2m long with climbing nature. Leaves are uniform and small with pine needle-like shape. Moreover, flowering season of Asparagus is between October and November. Later, it gives the attractive red berries like fruit [8,9].

15.3.2. Microscopic characterization

Microscopic section of the root tubers shows an outer layer of piliferous cells, ruptured at places and along with occasional unicellular root hairs. The cortex is distinguished into outer and inner cortex. The outer cortex consists of six or seven layers of compactly arranged, thick-walled, irregular to polygonal lignified cells. While the inner cortex is composed of 20–23 layers of thin-walled, oval to polygonal cells with intercellular spaces. Moreover, two or three layers of stone cells and raphides of calcium oxalate are also present in this region. The stone cells encircle the endodermis which consists of thin-walled parenchymatous cells. Below the endodermis, pericycle is present as a single layer of compactly arranged cell and the stele is exarch and radial in position. Xylem is composed of vessels, tracheids, and parenchyma and xylem vessels have pitted thickening. Phloem appears as patches alternating to xylem vessels. The pith comprises circular to oval parenchymatous cells with few lignified cells [10,9].

15.4. Cultivation of Asparagus racemosus

A. racemosus is a perennial plant generally distributed in tropical to subtropical regions up to an altitude of 2200m. It usually grows in a variety of soils, but sandy loam and soil rich in organic matter is more suitable for the growth of its root [11,8]. A soil with a pH of six to seven to slightly alkaline is favorable for higher yield, but it can be easily cultivated up to the pH 8. The roots are formed even in rocky soils with minimum soil depth of 30cm [12,13]. Additionally, well drained, black, and fertile soil are highly suitable for cultivation [14,8]. However, overwatering must be avoided. It mostly requires tropical, hot climatic conditions with a temperature of 25–40°C. Month of May or June is considered suitable for its harvesting, whereas for transplanting, July–August is favorable. In the month of July, it produces flowers which are minute, white, and unisexual. Moreover, it begins to bear fruits which are globular, pulpy, and turns to purplish black berries when ripened in September, and the seeds are hard and brittle [8,4]. The first harvesting is done after 1.5–2 years of transplanting, which can be continued for 10–15 years. Generally, the crops are not affected by pests and diseases. If seeds are required, male and female plants are grown. Furthermore, weeding operations are to be timely carried out [14].

15.5. Nutritional and traditional medicinal value of Asparagus racemosus

15.5.1. Traditional uses

Medicinal uses of Asparagus were firstly document in ayurvedic formulation. Traditionally, it has been used in various purposes for the treatment of several ailments such as diarrhea, piles, hyperacidity, bleeding disorder, gout, phthisis, duodenal ulcer, urinary disorders, and insufficient lactation [9]. In Bangladesh, A. racemosus is used for improving memory and cognition [15]. Moreover, the indigenous people in India use its root juice or powdered roots along with sugar, honey, or cow's milk for the treatment of epilepsy, leucorrhea and as a tonic andaphrodisiac [16]. Similarly, it has also been reported for its use as an antiseptic, anthelmintic, and antidiarrheal agent in different parts of India. In Nepal, people use decoction or juice from its roots for various genitourinary problems like burning micturition and gastrointestinal troubles such as constipation, diarrhea, dyspepsia, dysentery, laxative, etc. It is also used in diabetes, jaundice, and as galactagogue by women after delivery in order to promote lactation and as a tonic 10,17

In addition to the traditional uses, Ayurvedic literature explains several therapeutic benefits of this plant. Ayurvedic practitioners use its root as an appetizer, emollient, aphrodisiac, cooling, diuretic, galactagogue, rejuvenating, carminative, antiseptic, antispasmodic, hepatoprotective, and as tonic [12]. Its root is also claimed to have beneficial effects for nervous disorders, inflammation, fever, cough, bronchitis, dyspepsia, diarrhea, neuropathy, and hepatopathy [14,12]. Moreover, it is also used to prevent aging, improve immunity and mental function, and for the treatment of menstrual disorders [18]. However, it is not limited to the ayurvedic formulation, but Unani traditional medicine also uses Asparagus in the treatment of leucorrhea, dysentery, premature ejaculation, diarrhea, spermatorrhea, etc. [19,20].

15.5.2. Local food and other uses

People in Nepal consume its tender shoots and leaves as a vegetable and has a good demand and market value. The shoots are also eaten as pickle and soups in different parts of Nepal [21]. Its root is fed to the cattle for relieving milking disorder or to promote milk production. Some local people also use its root for fermenting local beer [5,17] . Furthermore, people also use its root for controlling fleas and for washing clothes [22].

15.6. Pharmacological activities

15.6.1. Antidiarrheal and antiulcer effects

The powdered dried roots of A. racemosus are recommended in Ayurvedic texts for the prevention and treatment of gastric ulcers, dyspepsia, diarrhea, and dysentery [12,23]. Ethanol and aqueous extracts of the roots showed antidiarrheal activity against castor oil-induced diarrhea model in rats [24]. The juice obtained from the fresh root of A. racemosus has been shown to have curative effect in patients with duodenal ulcers [23]. Moreover, A. racemosus when used along with Terminalia chebula was shown to reduce the severity of ulceration and ulcer index by protecting gastric mucosa against pentagastrin and carbachol induced ulcers [25]. Increased output of mucus has been suggested as the mechanism for its cytoprotective effect [26].

15.6.2. Galactagogue effects

The root extract of A. racemosus has been mentioned in Ayurveda as galactagogue and has been traditionally used to promote milk secretion during lactation [27]. Its alcoholic extract was reported to increase the weight of the mammary glands and maintained milk secretion in rats [28]. A randomized clinical trial reported that the powdered root of the plant increased the prolactin hormone level by three times as compared to the control group [29]. Its use in combination with other herbal substances in the form of “Ricalex” tablets (Aphali pharmaceutical Ltd., India) has been shown to increase milk production in females complaining of deficient milk secretion [30]. In another study, A. racemosus along with some other herbal substances in the form of a commercial preparation, lactare (TTK Pharma, India) is reported to enhance milk output in women complaining of scanty breast milk [12].

15.6.3. Phytoestrogenic effects

Phytoestrogens are the class of compounds obtained from the plants, which are both functionally or structurally similar to ovarian and placental estrogens and their metabolites and regulate the ovarian cycles in females [31]. Additionally, studies have pointed out that phytoestrogens can lower the incidence of breast cancer in women [32]. Ethanolic extract obtained from the rhizome of A. racemosus upon oral administration in female albino rats showed increase in body weight and considerable development of uterus and mammary glands. The study suggested that A. racemosus had an estrogenic effect on female genital organs and mammary glands [33]. A study on oral administration of a polyherbal preparation, Menosan (Himalaya, India) containing the extract of A. racemosus was reported to exert estrogenic activity by increasing the uterine weight and uterine glycogen level in immature rats without a change in estrogen and progesterone levels [34]. A clinical trial concluded that Menosan considerably improved the symptoms of postmenopausal syndrome without altering the serum luteinizing and follicle-stimulating hormone [35].

15.6.4. Antimicrobial effects

Methanolic extract of the roots of A. racemosus exhibited significant antibacterial activity against wide range of bacteria [36]. In another study, ethanol extract of the leaves and its fractions (hexane and chloroform fractions) were studied on different bacterial and fungal organisms. Ethanol extract was shown to have the highest antimicrobial activity followed by the chloroform extract, and least activity was observed in the hexane extract [37]. Moreover, high antifungal activity was observed from the methanol extract of roots and tubers against Candida [38]. Preliminary phytochemical investigations have indicated the presence of secondary metabolites such as glycosides, steroids, saponins, and flavonoids as active constituents for its antimicrobial properties [7].

15.6.5. Antiepileptic and neuroprotective effects

It has been found that methanol, chloroform, and aqueous extract of roots of A. racemosus were effective on the treatment of maximal electroshock (MES) and pentylenetetrazole-induced seizuremodel in rats [39]. Methanolic extract demonstrated significant anticonvulsant effect in rats in comparison to chloroform extract, aqueous extract, and control group. The authors indicated that higher antiepileptic effects of the methanolic extract of the plant could be due to the presence of steroids, saponins, and triterpenoids. Another study performed by Shastry and coworkers [40] evaluated the methanolic and ethanolic extract of root of A. racemosus on maximal electroshock (MES), picrotoxin, and strychnine-induced seizure models. The results showed that ethanolic extract exhibited considerable anticonvulsant effects in comparison with the methanolic extract.

In Ayurveda, A. racemosus is a well-known nervine tonic. It has been also observed that the root extract of A. racemosus considerably decreased the loss of neurons in hippocampus and medial prefrontal cortex in ovariectomized rats suggesting benefits of extract in improving cognitive function [41]. Similar results were obtained in another study where an ethanolic extract of the plant exhibited notable neuroprotection and improved learning and memory against ethanol-induced oxidative stress and cognitive impairment in albino male rats [42].

15.6.6. Immunomodulatory and immunoadjuvant effects

Diwanay and coworkers pointed out that extracts and formulations containing roots of A. racemosus showed various immunostimulatory functions such as increase in white blood cell counts, hemagglutinating and hemolytic antibody titers in cyclophosphamide-treated mouse ascitic sarcoma [43]. Similarly, oral administration of the decoction of powdered root of A. racemosus has been reported to produce leukocytosis and predominant neutrophilia along with enhanced phagocytic activity of the macrophages and polymorphs [12]. Alcoholic extract has been found to enhance both humoral and cell-mediated immunity of albino mice injected with sheep red blood cells as particulate antigen [44].

15.6.7. Antidiabetic activities

Ethanolic extracts of A. racemosus roots have been shown to enhance insulin secretion in perfused pancreas and isolated islets. The extract significantly suppressed postprandial hyperglycemia after sucrose ingestion and reversibly increased unabsorbed sucrose content throughout the gut. The extract also inhibited the absorption of glucose during in situ gut perfusion with glucose. Daily administration of A. racemosus to type 2 diabetic rats for 28 days decreased serum glucose, increased pancreatic insulin, plasma insulin, liver glycogen, and total oxidant status [45].

15.6.8. Lithiatic activity

It has been reported that calcium, phosphorus, urea, and other urolithiatic factors were significantly reduced in rats on treatment with 800 and 1600mg/kg of ethanolic extract. Similarly, the ethanolic extract of Asparagus inhibited stone formation via regulating the ions from urine on 0.75% ethylene glycol-induced urolithiatic models [41].

15.7. Chemical constituents

A. racemosus is shown to possess a diverse range of phytochemicals such as carbohydrate, flavonoids, alkaloids, phenolic compounds, steroids and/or terpenes, saponins, vitamins, and minerals [9]. Moreover, its root is reported to contain 36.8–52.89% carbohydrates, 2.95–6.1% protein, 17.93% crude fiber, 4.1–8.83% saponins, 5–6.2% oil, and 4.18% inorganic matter in addition to vitamins, flavonoids, and polyphenols [46].

Steroidal saponins (shatavarins) are the major bioactive constituents of A. racemosus. Shatavarin I and Shatavarin IV are the important steroidal saponins and glycosides obtained from the root of A. racemosus. Shatavarin I is the glycoside with three glucose and rhamnose moieties attached to sarsasapogenin. Whereas, Shatavarin IV is a glycoside of sarsasapogenin having two molecules of rhamnose and one molecule of glucose [47,7].

Several phenolic compounds called flavonoids have also been isolated from A. racemosus. Kaempferol, quercetin and glycosides of quercetin, such as quercetin-3-O-rutinoside (rutin) and quercetin-3-O-galactoside (hyperoside) have been isolated from roots and flowers, whereas quercetin-3-glucuronide has been isolated from leaves [48]. A novel antioxidant compound called racemofuran has also been isolated from the roots of A. racemosus along with previously isolated antioxidants asparagamine and racemosol [49]. The structures of representative compounds in A. racemosus are presented in Fig. 15.2.

A. racemosus is reported to contain several vitamins such as A, B1, B2, C, E, and folic acid. Moreover, minerals such as potassium, calcium, iron, sodium, copper, manganese, cobalt, zinc, copper, lithium, etc., have also been detected from different parts of this plant [8]. Another study investigated the polysaccharide composition of A. racemosus roots (MW, 2000kDa) and found high amounts of galactose and glucose and smaller amounts of arabinose and other sugars (galactose 54%, glucose 28%, rhamnose 4%, xylose 5%, and arabinose 8%) [48].

Figure 15.2
Representative structures of isolated compounds from Asparagus racemosus.

15.8. Clinical studies

A clinical study on oral administration of 2gm of powdered root of A. racemosus showed comparable gastric emptying time with metoclopramide, which supports its traditional use for treating dyspepsia [50]. A study performed by Mangal and coworkers to patients with chronic peptic ulcer demonstrated that 3g of the root powder of the plant with milk upon oral administration twice a day for six weeks relieved the symptoms associated with peptic ulcer [51]. Similar results were obtained in another study where the powdered root of the plant showed symptomatic improvement in patients with duodenal ulcers [52].

A double-blind, randomized controlled trial where the powdered root of the plant in the form of capsules (dose of 60mg kg
−1 body weight per day) administered orally to lactating mothers was performed to evaluate the galactagogue effects of A. racemosus. Oral ingestion of the capsules three times a day with milk for 30 days increased the prolactin hormone level by threefold in comparison to the control group with no signs of acute oral toxicity [29]. Another study revealed that the formulation containing the extract of A. racemosus increased milk production and promoted lactation in mothers with decreased milk output [30].

A study on patients undergoing treatment for stress-related problems like memory disorders, cognitive changes evaluated the neuroprotective effects of A. racemosus root extract. The study reported that the group receiving the plant extract exhibited a significant increase in test score (retention and recall test) [53].

15.9. Adverse effect and precaution

Although A. racemosus has high potential as remedy for various kinds of aliments, the toxic dose of Asparagus should be considered. Previously, it was reported that the 2000mg/kg extract of Asparagus also did not produce any significant change in the body, and other parameters after treated with certain enzyme. Similarly, ethanolic as well as aqueous extract of Asparagus also did exhibit acute or subacute toxicity in rat model at 2000mg/kg and 3200 mg/k, respectively. Interestingly, Asparagus can be used safely during pregnancy and lactation [54]. However, partial teratogenicity in rodents was observed in pre- and postnatal studies in alcoholic root extract of Asparagus at 100mg/kg/day for 60 days [55]. Asparagus is known to interfere the physiological function of the body, suggesting caution on using lithium supplements or lithium-containing medication [56,57]

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Chapter 16: Cinnamon as a potential nutraceutical and functional food ingredient

A. Thahira Banu, and Janeline Lunghar
School of Sciences, Department of Home Science, The Gandhigram Rural Institute-Deemed to be University, Dindigul, Tamil Nadu, India

Abstract

Cinnamon (Ceylon cinnamon/cassia) is a spice obtained from the bark of the cinnamon tree, a tropical medicine, belongs to the Lauraceae family. Cinnamon is used as a flavor enhancer in cooking since ancient time. Today, it is not only known as a flavored spice but also has gained tremendous applications in therapeutics. Research has proven the medical benefits in lowering cholesterol, blood glucose, and improving insulin resistance. Cinnamon has a unique aroma and pungency attributed to the compounds, cinnamaldehyde and eugenol. It contains polyphenols that regulate glycemic levels. An array of bioactive compounds in cinnamon is responsible for its antiinflammatory, anticancer, antibacterial, and antioxidant activities. This chapter focuses on the role of Cinnamon zeylanicum/cassia as a natural medicinal spice with its nutraceutical and functional food properties, mechanism of action, tolerable daily intake for healthy living, and traditional culinary preparations.

Keywords

Cinnamaldehyde; Cinnamon cassia; Cinnamon zeylanicum; Eugenol; Functional food ingredient; Therapeutic properties

16.1. Introduction

Functional foods and nutraceuticals are hitting the market, and the growing trend for this is due to extensive research in identifying its application and consumer awareness. According to the International Food Information Council, functional foods are “foods or dietary components that may provide a health benefit beyond basic nutrition” [1]. Nutraceuticals are isolated from the plant, animal, and microbial groups, and Health Canada states that nutraceuticals are a product that is “prepared from foods but sold in the form of pills or powders, or other medicinal forms not usually associated with foods.” It has been considered to provide physiological benefits and prevention against chronic diseases [2]. There is a galaxy of novel functional foods and nutraceutical components available in the market. Individuals use these for well-being, treatment of diseases such as cancer, diabetes mellitus, cardiovascular disease (CVD), Alzheimer's, and other degenerative diseases. Among them, spices harbor enormous nutraceutical properties and can behave as a functional food. Cinnamon is one of the essential spices that is known for its therapeutic and culinary use. Cinnamon is a spice obtained from the bark of the cinnamon tree, a tropical medicine, belongs to the Lauraceae family. There are more than 250 species known around the world. The most common among them is Cinnamomum verum/zeylanicum, Cinnamomum cassia, Cinnamomum burmannii, and Cinnamomum loureiroi. Cinnamomum tamala is a variety grown in the Himalayas at an altitude of 3000 to 8000ft in Assam and Sikkim, and the leaves of this species are commonly called bay leaves [3]. The major countries producing cinnamon are Sri Lanka, China, Indonesia, and Vietnam. The leaves and barks are used for essential oils and as flavoring agents. It has long been an ingredient in cooking meat, baking, and Ayurvedic medical preparations. Cinnamon is available as tubular rolls, also called quills, sticks, powders, and essential oils. The spice contains phytochemically active compounds with antioxidant properties. Furthermore, many current scientific reports found that cinnamon is used as an antidiabetic, antioxidant, antiinflammatory, antifungal, and anti‐HIV; improve heart function, pro‐wound healing, immune enhancers, and anticancer [4–7]. The following sections deal with the species, composition of phytochemicals, nutrients, therapeutic properties, safety, culinary uses, role as functional food components, and future research prospects. This chapter will focus on Ceylon cinnamon/Cinnamon zeylanicum and Cinnamon cassia.

16.2. Species

Discovery of modern antibiotics and antimicrobial agents, spices have existed in consumable form from ancient days as a resource for healing ailments. Cinnamon (Cinnamomum) is a spice obtained from the inner dried bark of several species of indigenous trees to South Asia, China, and belongs to the genus Cinnamomum. The history of cinnamon usage references is found in the scriptures [8]. From ancient days, cinnamon is a traditional medicine in treating many conditions, namely bronchitis, rheumatism, neuralgia, etc. The four significant species of cinnamon in the genus Cinnamomum are C. verum, C. cassia, C. burmannii, and C. loureiroi. C. verum is called Ceylon cinnamon, and its older botanical name is Cinnamomum zeylanicum. C. cassia is called Chinese cinnamon, and C. burmannii as Korintje, Java, or Indonesian cinnamon, and C. loureiroi is known as Vietnamese or Saigon cinnamon. Apart from this, there are approximately 250 species of cinnamon of lesser importance available around the world as a spice or medicinal preparations [9,10].

Approximately 250 species have been identified among the Cinnamon genus, with trees being scattered worldwide [11,12]. C. cassia is the common species available and is an evergreen tree that grows up to 20–30 feet; the leaves are dark green on top and lighter green underneath. Cinnamon fruits are black, pulpy, and aromatic. A flower is tiny in yellow color [13]. The barks of cinnamon species are pretty popular in cooking worldwide, and it is historically used in traditional and modern medicines. It is reported that Sri Lanka produced 90% of the world's true cinnamon, and this spice is derived from the brown bark, which forms quills with longitudinal striations. The spice plant is native to Sri Lanka, South-eastern India, and other native-like Indonesia, South America, and the West Indies [14–16]. The bark of the plants is used for spice and medical purposes [17,18]. Only one cinnamon is present and commonly referred to as Ceylon cinnamon, and it is from the plant Cinnamon zeylanicum. Cassia is not common, but it is marketed to consumers as cinnamon at a cheaper rate. The Ceylon cinnamon has a minimal amount of coumarin compared to cassia. Consuming cassia for a more extended period would result in health implications. The coumarin content should not be higher than 0.3% by weight [19,20].

The following are the differences listed in Food Safety Standards regulations (FSSAI, 2018) between Ceylon cinnamon and cassia.

- 1. The presence of more than 5%–6% of cinnamaldehyde in cassia add to its hot and robust flavor.

- 2. True cinnamon is light brown or tan, and cassia is dark brown or reddish-brown.

- 3. True cinnamon curls up from one side and roll up, and cassia looks like a bark stripped off a tree. Fig. 16.1 shows the difference between Ceylon zeylanicum and cassia.

- 4. The bark of true cinnamon is soft and smooth; cassia is rugged and uneven.

As stated earlier, true cinnamon is grown only in Sri Lanka and some parts of India, whereas cassia is grown in China and Vietnam. As per the German Federal Institute for risk assessment, onekilogram of Cinnamon cassia has approximately 2.1–4.4gm of coumarin [21]. The amount is higher than the tolerable daily intake (TDI). The TDI given by the European Food Safety Authority (EFSA) for coumarin is 0.1mg/kg/body weight/day [22]. Whereas the coumarin content of Cinnamon zeylanicum  is very low. Coumarin is a secondary group of phytochemicals with strong anticoagulant, hepatotoxic activities, so it is always recommended to use Cassia zeylanicum in food preparations as a functional food. Hence, in this chapter, all varieties of cinnamon are not highlighted, and the main focus is on Cassia zeylanicum (true cinnamon/Ceylon cinnamon/Cinnamon verum) and Cassia cinnamon.

Figure 16.1
Cinnamon ceylon zeylanicum and cinnamon cassia.

16.3. Composition of cinnamon

The main compositions of C. cassia are terpenoids, glycosides, phenyl propanoids, etc. However, cinnamon composition differs depending on multiple factors, which includes part of the plant, the growing season of the plant, age of trees, location, and method of extraction [23,24]. The active components in cinnamon (C. zeylanicum) are polyphenols and volatile phenols. Among these phytonutrients, cinnamon contains mainly vanillic, caffeic, gallic, ferulic, protocatechuic, and p-coumaric acids [25]. Regarding volatile components, the composition of cinnamon essential oils depends on the part of the plant extraction. In bark essential oil, cinnamaldehyde constitutes 73%–90% and this depends on the type of extraction process. Studies revealed that extraction using steam distillation provided higher concentration than Soxhlet extraction [26]. The other minor volatile compounds are β-caryophyllene, linalool benzyl benzoate, eugenyl acetate, and cinnamyl acetate. In regard to cinnamon leaf essential oil, the key component is eugenol, which reaches a concentration of more than 80%. Essential oil obtained from cinnamon fruit and flowers, the major components are (E)-cinnamyl acetate, caryophyllene, and phenolic compounds [27] and 74.92% contained eugenol [28]. The presence of cinnamaldehyde in C. zeylanicum is responsible for its antihyperglycemic [29,30], antioxidant [31], antiinflammatory, and anti-Alzheimer's effect [32,33].

C. zeylanicum Blume was tested for its phytochemical composition and reported the presence of trans-cinnamic acid, cinnamaldehyde, cadienene, terpinen-4-ol, β-caryophyllene, and coumarin [34]. The three main components of C. zeylanicum essential oil from bark namely, trans-cinnamaldehyde, eugenol, and linalool, which, according to Chericoni et al. represent 82.5% of the total composition [35]. The active compound of C. zeylanicum bark oil is, trans-cinnamaldehyde approximately 49.9% to 62.8% of the total amount [36]. Further, cinnamaldehyde and eugenol also are significant components of cinnamon extract. Four distinct components—cinnamaldehyde, cinnamic acid, cinnamyl alcohol, and coumarin were found in the dried stem bark of C. cassia. He et al. identified high contents of cinnamaldehyde (13.01–56.93mg/g) in C. cassia bark [37]. Interestingly, Friedman et al. found the presence of volatile compounds, eugenol and linalool are stable to heat, unlike pure cinnamaldehyde, which are not heat stable and undergo a temperature-dependent transformation to benzaldehyde under the influence of heat starting at approximately 60°C [38]; thus, addition of cinnamon should be at the end of cooking for enjoying its fullest benefits. Currently, there are studies on the evidence that C. cassia has more than 160 chemicals with pharmacological applications. Studies have confirmed that C. cassia has a wide range of pharmacological effects, including antiinflammatory, antitumor, antidiabetic, antibacterial, and antiviral, cardioprotective, neuroprotective, immunoregulatory effects, cytoprotective, neuroprotective, immunoregulatory effects, and antityrosinase activity [39,40]. Different chemical compounds of Cinnamon zeylanicum plant are listed in Table 16.1.

Table 16.1

Recommended dosage of Piper nigrum. Chemical compounds of Cinnamon zeylanicum plant.

S. No | Plants parts | Major compounds | References
1. | Bark | Cinnamaldehyde 65%–80% eugenol 5%–10% | Vangalapati et al. [12], Yanakiev [39], Tepe and Ozaslan [40]
2. | Root bark | Camphor 60% | Vangalapati et al. [12], Jayaprakasha et al. [41]

3.

Leaf

Eugenol 70%–90%, cinnamaldehyde 1%–8%
Cinnamaldehyde: 1.00%–5.00%
Eugenol: 70.00%–95.00%

Vangalapati et al. [12], Jayaprakasha et al. [41] Utchariyakiat et al. [42]

4.

Fruits

Trans-cinnamyl acetate 40%–50%, caryophyllene 10%–15%, trans-cinnamyl acetate (42.00%–54.00%), and caryophyllene (9.00%–14.00%)

Vangalapati et al. [12], Jayaprakasha et al. [41] Bakar et al. [43]

5.

Flowers

Trans-cinnamyl acetate 40%, trans-α-bergamotene 10%, caryophyllene oxide 8%, (E)-Cinnamyl acetate: 41.98%, trans-alpha-bergamotene: 7.97%, and caryophyllene oxide: 7.20%

Vangalapati et al. [12], Jayaprakasha [41], Bakar et al. [43]

6. | Buds | Terpene hydrocarbons: 78.00%, alpha-bergamotene: 27.38%, alpha-copaene: 23.05%, and oxygenated terpenoids: 9.00% | Vangalapati et al. [12], Jayaprakasha et al. [41]

16.4. Natural compounds of cinnamon and therapeutic uses

The fact is, cinnamon has plenty of varieties and its active ingredient is cinnamaldehyde. As cinnamon possesses medicinal compounds, it is widely used in flatulence, intestinal colic, diarrhea, digestive atony, etc. Also, for digestion, the essential oil can help in food processing by breaking down faster [44]. In individuals with type 2 diabetes mellitus, insulin is not produced adequately, thus glucose is not metabolized completely; studies indicated that supplementing cinnamon helps diabetics to metabolize glucose efficiently [45]. In connection to this, recent studies have reported that cinnamon helps to secrete insulin and metabolize glucose. Analysis of the insulin receptors (IRs) on exposure to the flavonoid methyl hydroxychalcone polymer isolated from cinnamon demonstrated the phosphorylation of the receptors and in turn triggered the insulin cascade. The MCHP is also referred to as mimetic to insulin and would help in the treatment of insulin resistance [46]. Additionally, the bioactive compounds present in cinnamon increased the receptivity of adipose cells to the insulin hormone that helps regulate the metabolism of glucose and thus controls the level of sugar in the blood.

Interestingly, cinnamon plant was identified with pharmacological properties [47]. Studies have revealed that the extraction of Cinnamomum plants can act against bacteria, diabetes, inflammation, tumor, and neuroprotective properties. Further, cinnamaldehyde works against the coagulation of platelets in the blood. It also hinders the release of arachidonic acid (a trigger for the inflammatory response) from cell membranes [48]. Bottom line, the cinnamaldehyde of cinnamon is helpful for inflammatory diseases like rheumatoid arthritis [49,50]. Further, recent studies showed that cinnamon aroma and scent act as cognitive enhancer that improve the brain performance, visual-motor capacity, and virtual memory [51] to its compound cinnamic aldehyde. Consumption of cinnamon reduces the levels of hemoglobin A1c (HbA1c) and fasting plasma glucose (FPG).

However, in contrast to this, a study reported that cinnamon consumption increases the level of FPG and HbA1c [52]. Nonetheless, effective doses are still not clear and deeper dive in is necessary. Randomized trials revealed a contrary result on intake of 1g/day [53], 1.5g/day [54], 2g/day [55], 3g/day [56], 5g/day [57], or 6g/day [58] showed different effects in glycemic and lipid parameters. However, this study did not indicate in regard to insulin resistance or sensitivity in acute doses of cassia [57]. Currently, cinnamon is available as a therapeutic supplement for many illnesses including various metabolic disorders such as arthritis [59]. Moreover, the plant is used as an effective medicine in various disorders in different regions of the world [60]. Cinnamon’s various parts are widely used in culinary applications such as liqueurs and chocolates as a flavoring agent [12,16]. It also reported that cinnamon has several applications in clinical problems as well as in therapeutics. It possesses antimicrobial, antioxidant, antiinflammatory, blood-purifying and blood-thinning properties, and it is also used in reducing blood sugar levels and cholesterols. However, excess intake of cinnamon can be harmful to body organs. Having said, many have investigated that cinnamon extracts and oil have antibacterial properties. Nimje et al. stated that the antimicrobial effect of chemical extracts of C. zeylanicum and C. cassia oil on three spoilage bacteria, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus [61]. Among these bacteria, the essential oil of C. cassia was found to have the best outcome on E. coli. According to the investigation by Ranjan et al., purified cinnamon extracts can be the best replacement for chemical preservatives [62].

A multitude of in vitro studies has demonstrated that cinnamon increases blood glucose uptake to cells by enhancing IR [63]. Cinnamon increases the amount of glucose transporter type 4 receptors as well as IR and IR substrate, thereby facilitating glucose entry into cells. Another possible mechanism for its hypoglycemic properties is an increase in the expression of peroxisome proliferator-activated receptor-α and peroxisome proliferator-activated receptor-γ, thereby increasing insulin sensitivity [64]. It has also been demonstrated that Ceylon cinnamon possesses an inhibitory effect on pancreatic amylase and intestinal sucrose [65]. Cinnamon also has the ability to increase glycogen synthesis and inhibit gluconeogenesis, by increasing the activity of pyruvate kinase and decreasing that of the phosphoenolpyruvate carboxy kinase [66]. On the other hand, the primary driving force for the potential of cinnamon bark for immune system benefits is polyphenols, especially proanthocyanidins. The true multifaceted clinical potential of cinnamon polyphenols has recent clinical evidence on various inflammatory conditions including allergic rhinitis and chemotherapy side effects. Evidence on cinnamon is the compounds of polyphenols can protect the immune system and use against chronic allergic rhinitis conditions, viral infections, and furthermore, to improve the quality of life by reducing the side effects of chemotherapy [67–69].

Cinnamon plants have been experimented and investigated as traditional medicinal plants, and it has pharmacological properties [47]. Many investigators said that the primarily focus on cinnamon's bioactivity is toward the recently generalized metabolic syndrome involving diabetes. Additionally, cinnamon medicinal plants have the potential and high efficacy against different bacterial pathogens without any side effects compared to antibiotics. Medicinal plants like cinnamon produce bioactive phytochemical metabolites such as flavonoids, flavones, flavonols, alkaloids, tannins, terpenoids, quinines, lectins, coumarins, and polypeptides. Moreover, cinnamon essential oils are considered good sources for new antibacterial agents and interact with bacterial cells using varied mechanisms of action [70]. The aqueous extract of CZ produced by high pressure methods showed the presence of bioactive compounds benzoic acid, (E)-cinnamaldehyde, trans-cinnamic acid, eugenol, cinnamtannin B1, and O-methoxy cinnamaldehyde. These compounds have the capacity to inhibit alpha glucosidase.

16.4.1. Antioxidant activity of cinnamon

True cinnamon possesses high antioxidant activity and is proven by many research studies. Chericoni [35], and Premakumara and Abeysekera [71] studied the antioxidant activity of C. zeylanicum extract taken from the bark and found that the major component was eugenol and cinnamaldehyde [35,71]. The bark of CZ Blume was evaluated for its bioactive compounds and reported that 74.92% is eugenol [31]. The other components responsible for antioxidant property has also been reported. The ethanol extract of C. zeylanicum contains caffeic acid, catechin, p-coumaric acid, p-hydroxybenzoic acid, rosmarinic acid, and quercetin [72]. The CZ Blume leaves collected from Gorakhpur, India, was tested for their phytochemical composition in the extracted oil and reported the presence of eugenol (87.3%) and others namely trans-cinnamaldehyde, and other compounds such as α-thujene, α-pinene, α-phellandrene, p-cymene, 1,8-cineole, β-caryophyllene, aromadendrene, germacrene-d, bicyclogermacrene, δ-cadinene, and spathulenol [27].

Aqueous, ethanol, acetone, and methanol extracts of CZ fruit had a high concentration of phenolic compounds responsible for its antioxidant activity [31]. The eugenol in the essential oil extracted from the bark of CZ helped decrease the 3-nitrotyrosine-induced lipid peroxidation (LPO) in in vitro studies [73]. The presence of volatile oils in cinnamon increased the antioxidant activity to 55.9% and 66.9% at 100 and 200ppm concentrations [74]. CZ bark extracts had potential radical scavenging activity of 65.3% of antioxidant activity [75]. Jeyaprakash et al. reported that the leaves, bark, fruit, and flowers of CZ contain 80 chemical constituents, and the major ones common to all parts were eugenol, camphor, and cinnamaldehyde. Trans-cinnamaldehyde acetate was found in fruits, flowers, and fruit stalks [76]. Bark and fruit contained proanthocyanidins. These compounds have double bis-flavan-3-ol units in molecule. All these compounds are responsible for the antioxidant, antimicrobial, and antidiabetic activities of CZ [31]. The research group reported that cinnamon oil has the inhibitory capacity to autoxidation of pyrogallol; hence, it is believed that it can mimic superoxide dismutase (SOD) activity [31].

Similarly, the cinnamaldehyde, cinnamic acid, and bioactive compounds present in cinnamon have proved to possess inhibitory action against nitric acid and expression of inducible nitric oxide as high as 81.5%, 71.7%, and 41.2% at concentrations of 1.0, 0.5, and 0.1 microgram/microliter, respectively [77].
Nanasombat et al. reported that essential oils of cinnamon showed high antioxidant activity. The antioxidant activity of CZ oil was determined and found to possess 61.46% antioxidant activity in β-carotene bleaching method, 2mM/mg reducing capacity in ferric reducing antioxidant power method, and 85% activity in the superoxide anion scavenging activity. The main compounds responsible for antioxidant activity are cinnamaldehyde, eugenol, and phenolic compounds. The essential oils are also reported to act as preservatives [78].

Ismail et al. analyzed the water extracts of five herbal plants. Among the plants, the CZ had the highest level of total phenols (270 GAE/100g) and flavonoids contents (60mg CE/100g). The antioxidant activity calculated for ABTS EC50 was 80% [79]. Phenolic compounds play an essential role to increase the antioxidant property of a food. The secondary bioactive compounds and their derivatives are also responsible for the antioxidant activity [80]. The phenolic compounds readily accept and donate electrons and hydrogen atoms and get paired easily with the free radicals that are oxidized. Further, they prevent free radicals from binding to metal ions that accelerate the LPO reaction. These mechanisms inhibit the free radicals or make it less hazardous and prevent LPO [81].

Dhuley [82] determined the antioxidant properties of CZ by feeding rats with a high-fat diet containing 10% cinnamon and measured the liver, cardiac antioxidant enzymes, and glutathione and lipid-conjugated dienes. The results indicated that the activity of the enzymes significantly increased, and the addition of this spice in food significantly activated the antioxidant enzymes [82]. Feeding rats with high fructose diet led to oxidative stress. The rats were supplemented with a high fructose diet with a spice mixture containing 1gm/100gm. It was believed that the spice mixture altered the level per oxidation markers in the tissues; the antioxidant status was improved in rats that received fructose diet along with spice mixture [83].

Ranjbar et al. supplemented 18 operating room personnel with CZ (100mg/300mL of tea) daily for 10 days and on analyzing the blood of the subjects for biomarkers of oxidative stress, namely LPO level, decreased total antioxidant power (TAP), and total thiol molecule (TMM) had no significant change [84].

Another study by the above researcher supplemented CZ (100 mg/30mL) of tea daily to 54 volunteers and observed the oxidative stress markers LPO level decreased, TAP and TMM increased in the supplemented group than the volunteers consuming plain tea [85]. Based on Wu et al., the United States Department of Agriculture released a list of oxygen radical absorbance capacity (ORAC) of selected foods. The lipophilic and hydrophilic ORAC was determined. Ground cinnamon had a considerable amount of lipophilic-ORAC and hydrophilic-ORAC of about 34.53μM of Trolox equivalent (TE)/g and 2640.83μMTE/g for lipophilic- and hydrophilic-ORAC, respectively. Total ORAC was 2675.36μMTE/g. Thus, cinnamons have a high potential antioxidant activity, add antioxidants to the diet, and act as a potential functional food ingredient in combating degenerative diseases [86].

16.4.2. Antidiabetic activity

The most recent statistical report on the prevalence of diabetes globally has estimated that 9.3% (463 million people) have diabetes mellitus and would tremendously rise to 10.2% (578 million) by 2030 and even higher with 10.9% (700 million) by 2045. The prevalence is comparatively higher in developed countries (10.8%) than in developing countries (7.2%). One in two (50.1%) people living with diabetes are not in the diabetic category [87]. The International Diabetes Federation stated that type 2 diabetes is associated chiefly with the sedentary lifestyle that leads to overweight. Besides, diabetes mellitus is an altered metabolism resulting from pancreatic β-cell dysfunction [46], less production of insulin or insufficient and the main target issues are insulin resistance and impaired glycemic index control [88]. These alterations lead to severe health complications. The bottom line in controlling postprandial glucose is pivotal to avoid complications [89]. Hence, food intake and physical activity are essential for preventing type 2 diabetes mellitus [90,91].

The array of hydrocarbon in different ratios is the same throughout the plant. This chemical architecture would be the main reason for the enormous beneficial medical properties. The mechanism by which it helps to reduce the blood glucose percentage reduces the intestinal absorption of glucose by inhibiting the hydrolyzing enzymes and enhances glucose uptake by tissues and glycogen synthesis, further inhibiting gluconeogenesis. The increase in insulin on the supplement of CZ is also another reason for lowering lipids. The high dietary fiber content of CZ also inhibits the absorption of lipid [16]. The bioactive compounds of cinnamon are involved in increasing the glycogen in the muscles and liver. Further, it inhibits the carbohydrate hydrolyzing enzymes alpha amylase and alpha glycosidase. Other probable mechanisms related to antidiabetic activity of cinnamon are stimulation of glycogen synthesis, uptake of glucose by cells by translocation of glucose transporters, reduction of gluconeogenesis, and increased glucose metabolism [29,92]. Cinnamon also can release insulin and increase IR activity [16,93].

Essential oil from C. verum reported to decrease the fasting blood sugar, cholesterol level, and urinary protein and alter the thiobarbituric acid reactive substances and catalase levels on administration to diabetic rats [94,95]. Further, in animal studies, it has revealed that C. verum can lower lipids profiles with increasing high-density lipoprotein (LDL) in hyperlipidemic individuals [16,94]. A study reported that, supplementation of C. cassia to sedentary obese women has remarkable reduction in glucose level [61]. Another study revealed that, cinnamon increases LDL levels but decreases the total cholesterol level, insulin, and triglyceride levels in type 2 diabetics [56]. Another similar study on the intake of C. verum extract for eight weeks reduces the serum ICAM-1 (association of intercellular adhesion molecule-1) level in patients with type 2 diabetes [96]. Cinnamaldehyde is the major compound present in C. zeylanicum essential oil, and it possesses antihyperglycemic activity, reduces blood cholesterol and triglyceride levels, and increases HDL cholesterol. The ethanol extract of CZ contains transabdominal preperitoneal-type A procyanidins polyphenols that inhibit carboxymethyl lysine formation in diabetic rat kidneys, modulated expression of renal markers, and improved proteinuria and diabetic nephropathy [30]. Lee et al. studied the glycemic control in diabetic patients supplementing cinnamon of 5, 10, and 20mg/kg of linalool chemotype had good glycemic control in people with diabetes due to secretion of insulin as supplementation reduced the oxidative stress and proinflammatory environment in the pancreas protecting beta cells of the pancreas [97]. Allen et al. conducted 10 clinical randomized controlled trials on supplementing 120mg/day to 6g/day of cinnamon for a period of four months to 543 patients. They reported a statistical decrease in fasting plasma glucose and reduction of triglycerides and lipids [98]. Alongside, many clinical trial studies on cinnamon are under process, and few completed studies reported the effectiveness of cinnamon supplementation in prediabetic patients by reducing plasma glucose and lipid levels in individuals, respectively [46].

16.4.3. Antiinflammatory activity

Inflammation is the natural way of body response to healing. It involves many chemical reactions that help to increase blood flow to a specific part of healing and generate pain as a signal that something is not healthy in the body [99]. Further, inflammation is pivotal because it is the pathophysiological process that support body homeostasis and repaired damaged tissues caused by various injuries or immune responses [100]. The same pattern of the inflammatory process involved in other disorders with proinflammatory mediators [101]. The present scenario against inflammation medications still relies on nonsteroidal drugs [32]. However, these drug classes can cause several side effects, including gastrointestinal ulceration. Therefore, attention has shifted toward natural and less side effect [100]. The use of natural compounds like cinnamon can successfully approach the treatment of several inflammatory disorders. CZ is known for its antiinflammatory effect against acute and chronic inflammation. CZ is studied for its hepatoprotective effects inducing a liver injury in rats through CCl_4 and treating with 0.01, 0.05, and 0.1g/kg for 28 days, and it was found that the toxicity was reduced and the serum liver marker enzymes alkaline phosphatase, alanine transaminase, and aspartate transaminase was positively altered SOD and catalase enzymes significantly increased in the treated rats [102].

The primary mechanism of the antiinflammatory activity of the cinnamon plant is the presence of cinnamaldehyde. It has indicated as one of the effective inhibitors expresses as NF-κB mediated by its antioxidant activity [103]. Further, cinnamaldehyde inhibits proinflammatory mediators such as interferons, interleukins, lymphokines, chemokines, eicosanoids (prostaglandins and leukotrienes), and reactive oxygen species involved in Alzheimer's disease [104]. Liao et al. reported in their study on the antiinflammatory effect of cinnamaldehyde on lipopolysaccharide (LPS) stimulated mouse macrophage (RAW 264.7) at 50μM [105]. A similar study reported that cinnamyl acetate revealed a significant antiinflammatory action on LPS-stimulated mouse macrophages (RAW264.7) [106]. Moreover, several flavonoid compounds (gossypin, gnaphalin, hesperidin, hibifolin, hypolaetin, oroxindin, and quercetin) have isolated and antiinflammatory activities [107–109 ]. A recent study reported that 2-hydroxycinnamaldehyde isolated from Cinnamon cassia bark exhibited an inhibitory effect on the production of nitric oxide by inhibiting the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), indicating that this substance can potentially be used as an antiinflammatory agent [110,111]. It has also reported that the ethanolic extract of Cinnamon cassia showed significant antiinflammatory effects by decreasing the activation of spleen-tyrosine kinase (Src/Syk-)-mediated NF-κB [110,112]. Furthermore, the aqueous extract of cinnamon reduces the LPS-induced tumor necrosis factor-α levels in the serum [8]. It was reported that, water extract of Cinnamon cassia decreased the TNF-α secretion in LPS-injected serum when administered orally for six days to mice. The results may be due to antiinflammatory property like polyphenol present in the extraction [8]. The ethanolic extract of Cinnamon cassia (100μg/mL) reported to have antiinflammatory potential; it can decrease NO production, TNF-α secretion, and prostaglandin E2 concentration in LPS-activated cells [112].

16.4.4. Antibacterial activity

As known, to suppress the microorganism growth, synthetic drugs have been adopted to inhibit the growth of pathogens [113–115 ]. However, it is pivotal to discover the alternatives to antibiotics such as natural antimicrobial compounds effective as the synthetic drugs [100,116]. The key potential mechanisms of antibacterial actions are related to the degradation of the bacterial cell wall with increased membrane permeability and the leakage of cytoplasm materials. Further, the natural compound of cinnamon plants does the shrinkage and distortion of bacterial cells and denature of bacterial protein [46].

Oussalah et al. evaluated the bacterial efficacy of cinnamon essential oils from species of Cinnamon cassia and Cinnamon verum against Pseudomonas putida, and it has been reported that minimum inhibitory concentration (MIC) values of 0.05% and 0.1% w/v, respectively [117]. In another study, the essential oils of Cinnamon cassia leaves of MIC were 750mg/ml for Salmonella typhimurium and 500ppm for E. coli, Bacillus cereus, and Lactobacillus sakei [118]. Further, methanol extract of Cinnamon cassia was also evaluated against other species of E. coli by the disk diffusion method and the microdilution, and it could inhibit microbial growth as it has antibacterial potency [108].

A study by Yap et al. with essential oils of Cinnamon verum bark against E. coli evaluating the permeability of its outer membrane, and it was observed that membrane damage was determined by denaturation and acidification of the cell membrane [119,120]. This suggested that the essential oil of Cinnamon verum bark disrupts the cell membrane by increasing the availability of the antibiotic in the bacterial cell [121]. Further, Cinnamon verum essential oils can inhibit S. aureus multidrug resistance. Moreover, cinnamon oil at concentrations of 3.12% highly inhibited the Streptococcus mutants isolated with the MIC values of 12.8–51.2 and 64–256 mg/ ml, respectively [122]. Rossi et al. revealed that Cinnamon verum essential oils could retard microorganism like Salmonella enterica, which involved the reduction of polyphenol oxidase activity [123]. Wu et al. also indicated that other species of cinnamon-like camphora essential oils had a potency to fight against E. coli with an MIC and minimum bactericidal concentration of 200μg/L [124]. The antibacterial effect is due to the presence of trans-cinnamaldehyde that inhibits bacterial acetyl-CoA carboxylase [125].

16.4.5. Anticancer activity

Cinnamon is a medicinal spice. Shefali et al. reported that cinnamon extract could cure cancers, including lung tumor cells, leukemia, ovarian and breast cancer, as it can inhibit the proliferation of cancer cells [126]. In addition, Thompson et al. revealed that cinnamon oil has anticancer properties due to its active components. Further, in vitro models have shown that the active constituent (trans-cinnamaldehyde) in cinnamon oil showed different tumorigenic and metastatic potentials [127]. Dutta et al. reported that the active component (trans-cinnamaldehyde) also showed effectiveness in breast cancer, subsequently suppressing cancer cells and migration in breast cancer representing various tumorigenic and revealed metastatic potential and components on oncogenic regulators and related pathways [128].

Cinnamon contains essential oils and various chemical constituents such as cinnamaldehyde, eugenol, benzyl benzoate, cinnamyl alcohol, cinnamic acid, cinnates, etc. It is used in the treatment of multiple diseases such as cancer, diabetes, inflammation, and microbial diseases [127]. Abak et al. reported that extraction of cinnamon (100μg) can inhibit the bacteria Helicobacter pylori, which is more effective than the commercial antibiotics as well as thyme herb extraction (200μg) [129]. Furthermore, the cinnamon essential oil can prevent cervical cancer as it suppresses and prevents from spreading into other areas. The main inhibition is from polyphenols and cinnamaldehyde and play a therapeutic role [130]. Several reports have dealt with the properties of cinnamon in different forms.
However, be it of bark, flower, powder, or essential oils. They are rich in flavonoids and phenolic compounds, and their active components are numerous as stated above. Thus, cinnamon plays a significant role in human health and has the properties of antimicrobacterial, antiinflammatory, anticancer, and antidiabetic.

16.5. Other therapeutic benefits

16.5.1. Antiulcer and gastric effect

CZ is used in the treatment of ulcers and other gastric problems. In vivo studies conducted in animals by Rao and Lakshmi proved that it reduced diarrhea in mice that was induced by using magnesium sulfate and feeding of 100 and 200mg/kg reduced the diarrhea by 71.7% and 80.4%, respectively [131]. The ability of cinnamon to inhibit H. pylori is its another mechanism in preventing cancer. It plays a role of antibiotic activity toward H. pylori. As the known H. pylori bacterium can raise the risk of various chronic health such as gastritis, peptic ulceration, and gastric cancer as well as coronary heart in humans. Furthermore, it has been reported that cinnamon extract (100μg) can inhibit H. pylori better than the commercial antibiotics to name few ampicillin (10μg), tetracydine (30μg), and cotrimoxazole (50μg), as well as thyme extract (200μg) [132].

16.5.2. Alzheimer's disease

Alzheimer's disease is caused by the degeneration of the neurons and by the formation of the senile plaque in the brain that contain amyloid beta peptide and hyperphosphorylated tau protein [133]. Polyphenols in CZ extracts are active and work against the activation of oxidative stress enzymes [134].

16.5.3. Parkinson's disease

One of the major neurological disorders is caused due to the degeneration of the midbrain, and this, in turn, reduces the hormone like dopamine in the brain, additionally, oxidative stress. The pathophysiology of the patients shows the presence of Lewy bodies from alpha-synuclein protein [135]. Various studies have indicated that sodium benzoate present in CZ exerts a neuroprotective effect [136].

16.5.4. Cardiovascular diseases and cancer

Cinnamon oil is used in the treatment of CVD. A rat study was conducted and animals were grouped in seven numbers into four groups. One group had healthy rats, second with diabetics and treated with probiotics, the third group with rats treated with cinnamon extract, and the fourth group with a mixed form of treatment. The rats treated with cinnamon along with food had reduced blood glucose as well as cholesterol and triglycerides. The possible mechanism is that CZ has a substantial amount of dietary fiber and could be a reason for the reduced lipid absorption [137]. The antioxidant activity of cinnamon can also be a reason for the low absorption of cholesterol. Insulin also has a role in reducing serum insulin levels, which in turn controls lipid metabolism [138].

16.6. Advanced glycation end products

Number of polyphenols and flavonoids are isolated from cinnamon bark, leaves, fruits, and buds such as catechin, epicatechin, and procyanidin B2 are isolated. These compounds inhibited the formation of advanced glycation compounds. These antiglycation activities of the phenolic compounds are attributed to their antioxidant activities. They are associated with the trapping capacity of reactive carbonyl species, namely methylglyoxal, an intermediate reactive carbonyl of advanced glycation end (AGE) formation. The ability to prevent AGE formation by entrapping the reactive carbonyl species has led to bioactive compounds in the treatment of diabetes and its complications [139,140].

Safety levels: Risk assessments of food ingredients for functional foods are performed to understand the tolerable safe limits and quality standards for safe consumption [141]. FSSAI, 2016 guidelines for nutraceuticals and functional foods have issued the permitted safety range of raw ingredients for adults per day for Cinnamon zeylanicum/Cinnamon verum is 1–2gms (not recommended during pregnancy) and has listed Cinnamon zeylanicum/C. verum as a nutraceutical ingredient [142].

The quality standards of C. zeylanicum/C. verum, also referred to as Dalchini for use is laid by FSSAI, and the following are the characteristics:

- ⁃ The powder should be yellowish to reddish-brown.

- ⁃ It must be with its characteristics flavor and aroma and free of off-flavor and mustiness.

- ⁃ As bark or powder should not contain more than 12% moisture by weight.

- ⁃ Volatile oil content not less than 7% by weight in bark and powder.

- ⁃ Total ash not more than 7% content by weight.

- ⁃ Insect damaged matter not more than 1% by weight.

Based on various studies and for thousands of years, cinnamon has been known as safe flavoring agent in food with no side effects [143,144]. According to the US Food and Drug Administration, cinnamon is generally recognized as safe in quantity found in food [145]. According to Khan et al. [58], frequent consumption C. cassia (6g per day) for 40 days has no harmful effect. Nonetheless, it has reported with mild gastrointestinal irritation, double vision, dizziness, and headache. As stated earlier in the chapter, TDI suggested by the EFSA for coumarin is less than 0.1mg/kg body weight [22].

16.7. Culinary uses of cinnamon across the globe

In India, spices and herbs play an important role in medicine and food preparations and possess potential medicinal benefits [146]. As per the Central Council for Research in Ayurveda and Siddha, 2005, it is advised to consume 1–2g Cinnamon verum bark powder in water for indigestion, vomiting, and nausea. Two grams of the bark powder in honey will help overcome the loss of appetite [147]. In general, cinnamon is used as a flavoring agent and has traditionally been used in Asian and Middle East culture [148]. In Egypt, a conventional drink is prepared using 4g of cinnamon bark powder in 200mL of water, boiling it below its boiling point and consumed as a beverage [149]. In a few countries, they are using cinnamon powder in all baked products. Famous brands are operating worldwide to name “CINNABON,” a renowned bakery operation in Dubai and Europe, and cinnamon flavored foods are becoming famous [150,151]. Meat cooked with cinnamon is an exotic food of Greece. The concentration of cinnamon used in food preparations is not scientifically validated [152].
Moreover, the spice merges well with chocolate and fruit dishes. Ground cinnamon is used in curry powders, masalas, and spice blends. Sometimes leaves are also used in baking trays to infuse the flavor and aroma in the baked products [153]. Thus, all the traditional recipes prepared with cinnamon can be tested in the perspectives of functional food properties.

16.8. Conclusion

Cinnamon’s research has moved miles from culinary use to functional properties to pharmaceutical, etc. They are economically important and broadly used worldwide. One of the major uses of cinnamon is its fragrance, and it is incorporated into various food products. Majorly, it is also a traditional natural medicine as cinnamon possesses many functional properties such as antidiabetes, antiinflammatory, antibacterial, and other serious illnesses. Nonetheless, further studies are necessary to determine the effectiveness of the active aesthetic of cinnamon and its specific part of the plant and their therapeutic effects. No doubt, numerous in vitro and in vivo studies have demonstrated an array of beneficial health impact in regard to diabetes, inflammatory disorder, and other chronic illnesses. But there is still a lack in regard to dosage and feeding regime. The authors noticed that there are limited publications in regard to the development of functional foods using bioactive compounds derived from cinnamon.

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Chapter 17: Myristica fragrans (Houtt.)

Arya Kadukkattil Ramanunny
[1]
, Sheetu Wadhwa
[1]
, Sachin Kumar Singh
[1]
, and Jithu Nambukulangara Vijayan
[2]

1
School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, Punjab, India
2
Department of Business Development, Zeon Life Sciences Pvt. Ltd, Sirmour, Himachal Pradesh, India

Abstract

Myristica fragrans (Houtt.) (M. fragrans) belonging to the Myristicaceae family is commonly known as nutmeg trees. The whole parts of the matured fruits such as rind, kernel (nutmeg), and mace (aril) are known for its flavor and fragrance. The odoriferous secondary metabolites such as volatile essential oil, fixed oil and oleoresin have commercial and medicinal value. Hence, in depth knowledge of the nutritional composition, chemistry of the phytoconstituents and its mode of action in exerting pharmacological activity are worthy contribution towards nutraceuticals and pharmaceuticals. This chapter emphasis the application of various parts of M. fragrans inclusive of rind, kernel and mace in a wide range of products such as culinary preparation, food and beverages, cosmetics, and pharmaceutical.

Keywords

Aril; Kernel; Lignans; Mace; Myristica fragrans; Nutmeg; Rind; Terpenes

17.1. Introduction

Myristica fragrans (Houtt.) (commonly known as nutmeg trees) belonging to the Myristicaceae family is a pyramid-shaped dioecious evergreen tree that usually grows up to a height of 20–25m [1]. The nutmeg trees are indigenous to Indonesia, specially to Banda Islands in Moluccas, which is also known as the Spice Island. Apart from this island, the species of Myristica genus are exotic to India, China, Western Sumatra, Grenada, West Indies, Northern Australia, Africa, Pacific Island, Jamaica, Nigeria, and Mauritius and Solomon Islands [2,3].

Nutmeg trees are mostly cultivated in hot humid climatic condition, and its plantation is possible in any kind of soil such as sandy loam, clay loam, or red laterite soil. It requires moderate rainfall, but care should be taken to avoid water clogging [4]. These evergreen trees have dark green-colored leaves with pointed tips and smooth surface. They are arranged alternately along the branches. It possesses bell-shaped pale-yellow flowers usually dioecious (male and female flowers are present on separate trees) and occasionally monoecious (both male and female flowers are together on a plant). The bark is noticed with pink sap. The ripe fruits are fleshy drupe that appears pale-yellow in color resembling an apricot [5].

The tree starts bearing fruits mostly in its seventh year and continues to do so throughout the year for approximately 30 years. However, the harvesting is done in April and November [6]. When the rind of the mature fruits is split opened, then there appears fleshy bright scarlet red aril, which surrounds the purplish-brown seed kernel. Generally, the inner dried seed kernel is known as nutmeg, and the red lacy (aril) covering the kernel is known as the mace [7]. Both nutmeg and mace are products from a single fruit and used as spices due to their similar flavor and fragrance. However, nutmeg is more preferred in culinary preparations by virtue of its aromatic, pungent fragrance, and slightly sweeter taste than mace. Besides nutmeg and mace, many other odoriferous secondary metabolites such as volatile essential oil, fixed oil (nutmeg butter), and oleoresin are obtained from its fruit parts and have commercial and medicinal importance. Traditional system of medicine uses nutmeg for the treatment of gastrointestinal problems such as flatulence, indigestion, colic, diarrhea, carminative, and stomachic. It is also used as a tonic, nerve stimulant, astringent, skin infections, rheumatism paralysis, and for the treatment of psychological disorders [8,9]. Thus, in this chapter, focus is given to the nutraceutical importance of M. fragrans and its fruit parts, emphasizing its functional components and explored pharmacological activities.

17.2. Taxonomy

More than 150 species are known under the genus Myristica. Some of the species that are grown in tropical regions are Myristica malabarica (Indian), Myristica argentea, and Myristica fatua. The diversity in species can be explained with the help of taxonomical classification [10,11].

Kingdom: Plantae
Division: Magnoliophyta
Class: Magnoliopsida
Subclass: Magnoliidae
Order: Magnoliale
Family: Myristicaceae
Genus: Myristica
Species: Myristica fragrans Houtt

17.3. Morphology and pharmacognostical details of M. fragrans fruit

M. fragrans fruit is a fleshy drupe with three distinct portions (Fig. 17.1):

1. • Rind: Thick fleshy pericarp, which appears in yellow color when ripe and resembles to an apricot.

2. • Kernel/nutmeg: Hard seed, which appears in ovoid or ellipsoidal shape and purplish brown in color.

3. • Mace/aril: Repeatedly lobed bright scarlet red flat ribbons, which loosely adhere on the seed but only fused at the base where it gets attached to the seed [12].

Figure 17.1
(A) Myristica fragrans tree bearing fruits, (B) Rind, (C) Ripe fruit split-opened and mace is visible and (D) Red mace and nutmeg seed removed from fruit rind.

Pharmacognostically, outer rind is known as pericarp, which comprises outer epicarp formed by a single layer of narrow, oblong thick-walled cells with prominent cuticle. Middle layer is highly vascularized mesocarp, which comprises parenchymatous polyhedral cells with numerous collateral vascular bundles which appear as single strand or in clusters. Inner endocarp comprises smaller elliptical thin-walled compressed cells.

Kernel/nutmeg is the seed which forms the outer seed coat (endocarp), comprises small parenchymatous cells with prominent cuticle (outer epidermis). Inner seed coat comprises radially elongated palisade like thin-walled cells with prominent vascular strands. Outer perisperm of the seed is formed from meristematic layer of angular parenchymatic cells, and inner perisperm has central core with parenchymatous cells and ingrowth are cylindrical lobed and vermiform. Perisperm is the nutritive tissue that surrounds the embryo. In between perisperm and embryo, soft thin-walled parenchyma cells are arranged to form endosperm. In young seeds, endosperm forms intrusion into the perisperm forming radial lobes (ruminant endosperm), but in matured seeds, endosperm disintegrates and loses its cellular content to make the ingrowth into a prominent perisperm.

Aril comprises epidermal layer of thin-walled cell having prominent cuticle. In the ground tissues, small and larger angular-shaped parenchyma cells are present in between the idioblasts. Vascular bundles are arranged in the median portion of the aril [12].

17.4. Estimation of composition in rind, nutmeg, and mace

All the three parts of M. fragrans fruit such as rind, nutmeg, and mace are widely used in the food industry due to its aromatic flavor and nutritive value. The biochemical estimation of each part of the fruit provides detailed information related to its content and percentage yield.

Gopalakrishnan in 1992 reported general composition of M. fragrans inclusive of rind, kernel (nutmeg), and aril (mace) as shown in Table 17.1. The volatile and nonvolatile substances were extracted from the kernel and mace. However, the kernel has reported highest concentration of nonvolatile components and mace has reported highest yield of starch and protein content [13].

Raj et al. in 2016 have carried out the biochemical estimation of rind content (Table 17.2) of 17 selected morphotypes collected from central Kerala [14]. The average moisture content in the rind was found to be 88%, which was slightly higher than the reported moisture value of 86% [15]. The titrable acidity of the rind has shown a protective action against microbial growth. Later, the antimicrobial activity of the rind has been explored in many value-added nutritional as well as medicinal products [14].

17.5. Phytochemistry of M. fragrans fruit

Plant and plant-based products are currently used in research studies owing to their phytochemical diversity. Understanding the chemistry of phytoconstituents and its mode of action in exerting pharmacological activity is a worthy contribution toward nutraceuticals and pharmaceuticals.

Phytochemicals are a heterogeneous class of compounds having structural diversity and variety of biological activities. Most of the phytochemicals are synthesized in plants through their biosynthetic pathway to obtain secondary metabolites such as the shikimic acid pathway [16] and acetate-mevalonate pathway [17]. Alkaloids, glycosides, phenolic compounds, terpenes, saponins, tannins, etc., are some of the biosynthesized secondary metabolites. Gayathri et al. in 2015 reported the phytoconstituents present in seeds, nutmeg butter, and mace of M. fragrans obtained by Soxhlet extraction. The phytochemical screening test of both aqueous and acetone extracts identified several phytoconstituents as shown in Table 17.3. All the extracts reported phenolic compounds and the total phenolic content varied from 7.3 to 14.2 mg/GAE g. However, higher phenolic content of 14.2 mg/GAE g was reported in the acetone mace extract [18].

Table 17.1

Composition of Myristica fragrans fruits [13].

Composition (%) | Rind | Nutmeg | Mace
Moisture∗ FWB | 88.00 | 40.00 | 40.00
Acidity | 20.83 | - | -
Volatile oil (v/w) | 0.83 | 11.00 | 15.30
Nonvolatile ether extract | 3.75 | 33.60 | 21.98
Starch | 0.42 | 30.20 | 44.05
Sugars
Glucose | 7.50 | 0.10 | 0.17
Fructose | 5.66 | 0.07 | 0.10
Total reducing sugars | 13.16 | 0.17 | 0.27
Sucrose | 0.75 | 0.72 | 0.39
Total sugars | 13.92 | 0.89 | 0.65
Protein | 5.3 | 7.16 | 9.91
Crude fiber | 27.58 | 11.70 | 3.93
Total ash | 7.42 | 2.57 | 1.56
Ash insoluble in hydrochloric acid | 0.58 | 0.20 | 0.15
Polyphenols
Total tannins | 2.42 | 2.50 | -
True tannins | 0.92 | 1.00 | -
Pectin | 14.10 | - | -

∗
FWB: Fresh weight basis. Except for moisture, all other components are evaluated on dried weight basis.

Table 17.2

Biochemical composition of fruit rind [14].

Composition | Limits
Moisture (%) | 87.1–89.4
Acidity (titrable acidity, %) | 1.28–1.92

Ascorbic acid (mg 100g
−1)

4.5–9.5

Pectin (calcium pectate, %) | 0.21–1.08

Protein (g 100g
−1)

0.21–1.85

Starch (g 100g
−1)

0.30–1.23

Total phenol (mg 100g
−1)

27.8–57.6

Tannins (mg 100g
−1)

143.3–750

Total minerals (%) | U2.01–2.57
Crude fiber (%) | 2.06–3.65

Moreover, the biodiversity and alterations in the geographical distribution have reported variations in the secondary metabolites and its yield. The majority of the investigations are focused on M. fragrans seed and mace. The extraction and identification studies of M. fragrans seed and mace have reported major phytochemicals such as phenolic compounds lignans, neolignans, phenylpropanoids, diphenyl alkanes, alkanes, fatty acids, fatty acid esters, terpenes, and terpene derivatives. Along with these major compounds, traces of steroids, saponins, triterpenoids, flavonoids, etc., are also reported.

17.5.1. Phenolic compounds

Phenolic compounds are secondary metabolites that are biosynthesized in plant tissues by shikimic acid and phenylpropanoid pathways. Chemically, an aromatic ring with one or more hydroxyl substituents constitutes them into several classes such as flavonoids, phenolic acids, tannins, stilbenes, and lignans. Compounds with a phenyl ring (C6) and side chain (C3) are known as phenylpropanoid, which serve as a precursor for the synthesis of other phenolic compounds as well as possess bioactivity.

Lignans and neolignans are high-molecular weight polyphenolic compound that are derived from shikimic acid biosynthetic pathway. The skeleton of lignans contains phenylpropanoid dimers, i.e., two units of C_6–C_3 molecule with a propyl side chain at C_2 position. Basically, tail-to-tail coupling of 2 phenylpropanoids occurs to form lignans. While neolignans are formed by head-to-tail or head-to-head coupling of phenylpropanoids, and hence the propyl side chain attachment in neolignans is not restricted to C_2 position [19]. The chemical structure of the lignans and neolignans obtained from M. fragrans seeds and mace is represented in Fig. 17.2.

Phenylpropanoids are phenylalanine-derived secondary metabolites that are derived from shikimic acid pathway. The structural diversity of phenylpropanoids due to enzyme transformations resulted in a variety of precursor molecules. Many derivatives are obtained during transformation where phenylalanine chain is substituted with one, two, or more aromatic rings, each ring with a characteristic substitution pattern as well as modification in the propane side chain of phenylalanine. The pharmacological activities of M. fragrans are exerted by some phenylpropanoids such as myristicin, elemicin, eugenol, and safrole, and their chemical structure is represented in Fig. 17.3.

Another chemical moiety found in M. fragrans whole fruits, seeds, and mace are diphenyl alkanes such as malabaricone C and malabaricone B, illustrated in Fig. 17.4.

17.5.2. Terpenes

Terpenes are the class of secondary metabolites, which are known to have isoprene unit. These are biosynthesized from mevalonate pathway (isoprenoid pathway) with the help of terpene synthase enzymes and cytochrome P450s [17]. Structurally isoprene units are 5 carbon atoms with a double bond, which form the basic skeleton of terpenes. Based on the number of isoprene units, terpenes are classified into monoterpenes (2 isoprene unit), sesquiterpenes (3 isoprene unit), diterpenes (4 isoprene unit), and triterpenes (6 isoprene units). Mostly essential oils of nutmeg and mace contain monoterpenes, sesquiterpenes, and oxygenated terpenes. Some terpenes and terpene derivatives obtained from oleoresins and essential oils such as α-pinene, β-pinene, sabinene, camphene, terpinen-4-ol, limonene, p-cymene, γ-terpinene, β-phellandrene, α-terpineol, geraniol, and linalool are shown in Fig. 17.5. These metabolites exhibited numerous pharmacological activities, which include antimicrobial, antiinflammatory, anticancer, and antidiabetic activity [19].

Figure 17.2
Structure of lignans and neolignans.

Figure 17.3
Structure of phenylpropanoids.

Figure 17.4
Structure of diphenyl alkanes.

Figure 17.5
Structure of terpenes and their derivatives.

Table 17.3

Phytoconstituents of Myristica fragrans seed, nutmeg butter, and mace in acetone and aqueous extracts.

S. No | Phytochemical test | Phytoconstituents | Acetone extract (seed) | Aqueous extract (seed) | Acetone extract (nutmeg butter) | Aqueous extract (nutmeg butter) | Acetone extract (mace) | Aqueous extract (mace)
1. | Mayer's test | Alkaloids | + | - | - | - | ++ | +
2. | Copper acetate | Diterpenes | + | + | + | - | - | +
3. | Ammonia | Flavonoids | ++ | + | - | - | ++ | +
4. | Acetic anhydride | Glycosides | + | - | + | - | + | ++
5. | Ferric chloride test | Phenolic | + | + | + | + | ++ | +
6. | Hydrochloric acid (HCl) | Phlobatannins | - | - | - | - | + | -
7. | Concentrated HCl | Quinones | + | - | + | + | + | -
8. | Frothing test | Saponins | - | + | - | - | - | -
9. | Liebermann–Burchard's test | Steroids | ++ | + | + | - | + | -
10. | Ferric chloride test | Tannins | + | - | + | - | + | -
11. | Salkowski test | Terpenoids | ++ | - | + | + | + | -

17.6. Extraction and analysis of bioactive components from M. fragrans

The extraction of bioactive components from M. fragrans has been reported with conventional extraction technique, advanced/emerging extraction technique, and hydrodistillation using Clevenger apparatus. Mostly, nonpolar solvents such as chloroform, petroleum ether, hexane, acetone, and ethyl acetate are used to separate out nonpolar components, whereas polar components are separated using polar solvents (water, ethanol, and methanol). Advanced technologies have utilized ultrasound waves/microwaves to enhance the yield of phytoconstituents with minimum extraction time and production cost [20,21]. Another method is supercritical fluid carbon dioxide extraction, an eco-friendly technique which reduces solvent usage, its disposal, and minimizes thermal degradation; hence, the phytoconstituents retain their chemical nature without any modification.

Secondary metabolites obtained from nutmeg extraction and distillation are subjected to analysis using various methods of detection. Generally, nutmeg has oils or volatile components; hence, commonly utilized technique is gas chromatography (GC) and GC with mass spectroscopic detections (GC-MS). Whereas, nonvolatile secondary metabolites are analyzed using high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC). Fingerprinting of methanolic extract of nutmeg seed was carried out for the identification of myristicin using normal-phase HPTLC with aluminum-packed silica gel 60F_254 as stationary phase and toluene: ethyl acetate (9:1) as mobile phase [22]. Chiu et al. in 2016 performed reverse-phase HPLC fingerprinting technique for extraction, fractionation, and purification of nutmeg kernels. Both volatile and nonvolatile constituents were isolated and later nuclear magnetic resonance technique was used for the identification of the 13 isolated compounds [23]. Previously, Ehlers et al. in 1998 developed reverse-phase HPLC technique for simultaneous determination of nine phenylpropanoids obtained from supercritical fluid carbon dioxide extract of nutmeg kernel and mace [24].

When nutmeg seeds are subjected to distillation, 5%–15% volatile oil [25] and 25%–30% fixed oils (nutmeg butter) were obtained. The major class of phytoconstituents obtained from both nutmeg and mace oil is monoterpene, sesquiterpenes, trace amount of oxygenated monoterpenes, and phenyl propene (aromatic ethers). Among the monoterpenes, β-pinene, sabinene, camphene, p-menth-8-en-ol, limonene, β-terpineol, and terpinen-4-ol are reported as major constituents and some of the sesquiterpenes are germacrene D and β-bergamotene [26,27]. Myristicin, safrole, elemicin, isoelemicin, eugenol, and isoeugenol are aromatic ethers that provide flavor, aromatic fragrance, and medicinal value to the oils [9]. Mallavarapu and Ramesh in 1998 have performed GC analysis of both nutmeg and mace oil and reported similarity in the composition with only variation in the quantitative estimation of the components [28]. Higher content of monoterpenes was reported in nutmeg oil (76.8%) than in mace oil (51.2%). However, oxygenated monoterpenes (51.2%) and aromatic ethers (18.8%) were higher in mace oil.

GC-MS analysis of essential oil obtained by hydrodistillation of the mace has reported 24 constituents. Among them, 11 were monoterpene hydrocarbon, predominantly sabinene was found at a concentration of 38.37% followed by other constituents such as α-pinene, β-pinene, 3-carene, 4-carene, β-myrcene, β-phellandrene, and D-limonene. Along with terpenes, distilled oil contained 12.43% of phenyl propene, which composed of myristicin (5.90%) and safrole (3.90%) [29].

A distinction in the phytoconstituents of kernel (without shell) and seed (with shell) were reported by Ashokkumar et al. in 2020 [29] for which hydrodistillation of kernel without shell and seed with shell were carried out. Later GC-MS analysis of the essential oil of the kernel without the shell has reported 24 constituents of which 12 were monoterpene hydrocarbons with total concentration of 89.41%. Sabinene (38.04%), α-pinene (19.22%), β-pinene (14.91%), D-limonene (7.52%), and β-myrcene (3.37%) were reported to be present in higher concentrations. Other constituents such as oxygenated monoterpenes (4.73%), sesquiterpenes (2.89%), oxygenated phenyl propenes (1.96%), and aromatic hydrocarbons (0.72%) were also present in trace amounts in the essential oil. Similarly, analysis of the seed with the shell has reported 25 constituents, out of which 13 were monoterpene hydrocarbons with a total concentration of 84.11%. The constituents present in kernel and mace oil are almost the same with variation in their concentration. The mace oil is rich with sabinene (27.65%), α-pinene (21.78%), β-pinene (18.23%), D-limonene (6.35%), and β-myrcene (2.91%). Along with these components, 4 phenyl propenes such as cis-asarone, myristicin, methyl eugenol, and safrole were also identified. The uniqueness of seed shell essential oil is the presence of α-bergamotene (0.63%), a sesquiterpene which is present in the seed shell only and hence not detected in any other parts of M. fragrans [29].

Matulyte et al. in 2019 reported a modified ultrasound bath-assisted extraction and hydrodistillation method with the addition of magnesium aluminometasilicate. The modified method yielded new compounds such as γ-amorphene, cis-α-bergamotene, β-copaene, bergamotene, citronellyl decanoate, cubebol, cubenene, and orthodene. The use of magnesium aluminometasilicate increased the quantity of sabinene from 6.53% to 61.42% and limonene from 0% to 5.62% [30]. Similarly, hydrodistillation of M. fragrans seeds from Grenada was carried out with a mixture of nutmeg powder, distilled water, and magnesium aluminometasilicate. This method also reported the presence of terpenes such as sabinene, α-pinene, β-pinene, D-limonene, γ-terpinene, α-thujene, camphene, α-terpinolene, γ-amorphene, α-bergamotene, isogermacrene D, α-copaene, and isoterpinolene. Along with these monoterpenes and sesquiterpenes, 2% myristicin (aromatic compound), 6% α-terpinyl acetate, bornyl acetate, less than 1% thujanol, and cis-p-menth-2-enol were present. But linalool and geraniol were not identified in this Grenada variant. However, the major components reported in the essential oil remained the same with a slight variation in their concentration [31].

17.7. Application of M. fragrans

The whole fruit inclusive of rind, kernel, and mace of M. fragrans has found its application in a wide range of products from various industries such as culinary preparation, food and beverages, cosmetics, and pharmaceutical. Therefore, each part of the plant is collected with utmost care. Once the fruits are matured and ripened, they may either drop down to the ground or are picked from the tree with the help of a hooked bill. After collection, the pericarp of the fruits is removed and used in culinary preparation like “pickles”. Then, the red mace present on the seed coat is carefully hand-picked, washed, flattened, and subjected to drying until they become brittle. The drying methods are carefully chosen to reduce the loss of valuable volatile components. Mostly, whole seed is dried under the sun until the inner seed produces a rattle sound upon shaking. The hardest seed coats are mechanically removed to collect the inner seed. Then, these dried nutmeg and mace are packed in double-layer packing materials, most preferably linen or polythene bags for retail sale or export [32].

A significant rise in the utilization of nutmeg, mace, and their derived products such as oleoresins, nutmeg butter, and essential oils in food, personal care products, and pharmaceutical industry has been noticed which enhanced their market value and demand. A detailed summary of their applications is described in the following sessions.

17.7.1. Source of nutrition

Spices are an important part of nutrition, and they also impart flavor to the dishes. Spices have the ability to improve and maintain human health and also prevent alterations in the physiological, metabolic, and psychological status [33].

Nutmeg is a source of energy, carbohydrate, protein, and dietary fiber. It contains both micro- and macronutrients such as vitamins (A, C, E), electrolytes (Na, K), minerals (calcium, copper, zinc, phosphorous, magnesium, manganese, iron), and other phytonutrients (β-carotene and β-cryptoxanthin). Recommended dietary allowance of each nutrient of nutmeg per 100g is given in Table 17.4 [34–36].

Table 17.4

Nutrient value of Nutmeg.

Dietary components | Nutrient value per 100g | Percentage of RDA for sedentary men | Percentage of RDA for sedentary women
Energy level | 525Kcal | 24.88 | 31.62
Carbohydrates | 49.29g | 37.91% | 37.91%
Proteins | 5.84g | 10.81% | 12.77%
Total fat | 36.31g | 145.24% | 181.55%
Dietary fiber | 20.8g | 69.33% | 83.2%
Micronutrients vitamins
Folates | 76μg | 25.33% | 34.54%
Niacin | 1.299mg | 9.27% | 11.80%
Pyridoxine | 0.160mg | 8.42% | 8.42%
Thiamine | 0.057mg | 4.07% | 4.07%
Vitamin A | 102 IU | 3.0% | 3.57%
Vitamin C | 3mg | 3.75% | 4.61%
Electrolytes
Sodium (Na) | 16mg | 0.8% | 0.8%
Potassium (K) | 350mg | 10.0% | 10.0%
Minerals
Calcium (Ca) | 184mg | 18.4% | 18.4%
Copper (Cu) | 1.027mg | 60.41% | 60.41%
Iron (Fe) | 3.04mg | 16% | 10.48%
Magnesium (Mg) | 183mg | 41.59% | 49.45%
Manganese (Ma) | 2.90mg | 72.5% | 72.5%
Phosphorus (P) | 213mg | 21.3% | 21.3%
Zinc (Zn) | 2.15mg | 12.64% | 16.28%
Phytonutrients
β-Carotene | 16μg | - | -
β-Cryptoxanthin | 90μg | - | -

∗Percentage RDA were calculated as per revised ICMR (2020) nutrient requirements for Indians [35].

In addition to its nutritional values, nutmeg has been extensively used as a flavoring agent. Both nutmeg and mace have shown delicate flavor; however, dried whole kernels are preferred over powdered forms as they possess more essential oils and give flavor to the recipes. At the last stage of cooking, dried kernels are added into the recipe either grated or milled to preserve the freshness of fragrance. Instead of ground nutmeg, essential oils are widely used as a flavoring agent to avoid residues in food and beverages. Natural preservatives are always in demand in comparison to synthetic ones due to their potential in reducing health risks as well as prevent foodborne diseases that are emerging health problems in the growing population. Nutmeg oil, oleoresin extracts, and nutmeg butter are some of the preservative agents of nutmeg that are included in the food item. In one of the study, antioxidant and antimicrobial activity of nutmeg essential oil has shown its application in biopreservation. These agents are found to be effective in retarding microbial growth and rancidity of food due to oxidation. Thus, food industries have high desirability to employ natural oils as preservative instead of synthetic preservatives [34].

17.7.2. Culinary and HORECA products

Nutmeg and mace are mostly used in a grounded form to give out their distinctive flavor, taste, as well as unique color. The sweet warm taste and pleasant fragrance of nutmeg are well utilized in many food products such as confectioneries (candy), puddings, meat and meat products (sausages), savory dishes, soups and sauces, baked food (cakes), desserts, masala tea, garam masala, range of alcoholic beverages to soft drinks, and mulled wines [37].

A chewable candy “Manisan pala” (sweetmeat nutmeg/fragrant fruit candy) is made with fresh pericarp and sugar syrup. The juice made from fresh pericarp is commonly available in Indonesia, Malaysia, India, and Caribbean. Mostly pericarps are used in countries where they are grown. The fresh pericarp is separated immediately after harvesting and processed for making the food items such as beverages, pickles, and chutney. It has been widely used in making jams and jellies in Caribbean and Far East Countries. In India and Middle East, nutmeg is used to flavor sweet dishes, whereas Europeans use them in soups, sauces, eggs, potatoes, meat, spinach etc. “Tempeh sausage” is a Javanese dish made from fermented soybean, which is prone to microbial growth upon storage. Addition of nutmeg seed extract improved the shelf life of the sausages by exerting antioxidant and antimicrobial activity, thereby maintaining color, texture, flavor, and moisture content of sausages even after storage [38]. Ben Lagha et al. in 2020 [39] evaluated the effect of methanolic extract of nutmeg on microbial growth in raw beef during its storage in chilled and frozen conditions. Nutmeg extract at 1.25% exhibited antibacterial activity against 8 foodborne pathogens such as Bacillus subtilis, Bacillus cereus, Escherichia coli, Klebsiella pneumonia, Listeria monocytogenes, Pseudomonas aeruginosa, Proteus mirabilis, and Staphylococcus aureus when stored at chilled (4°C) and frozen (−18°C) temperatures. This activity confirmed the antibacterial potential of nutmeg extract; hence, it can be utilized as a natural preservative [39].

Another innovative technique to delay the normal decomposition of the meat upon storage was done by wrapping them in an active film of M. fragrans. Galeano et al. in 2018 developed an active film containing nutmeg oleoresins at varying concentrations (0.5%, 1.0%, and 1.5%), gelatin (3%), polyvinyl alcohol (5%), and sorbitol solution (15%). The formulated active films were used for the storage of bovine loins (Longissimus doris) at 4°C. During storage, the total volatile basic nitrogen in the bovine loins was increased (<20mg/100g), which was less compared to the film without nutmeg oleoresin. This indicated the inhibitory effect of the active film against secondary oxidation. The delayed release of nitrogenous substances and oxygen barrier property of the active film was able to extend the shelf life of bovine loins [40]. Similar to this study, a biodegradable film of gelatin and chitosan with various essential oil was listed for storage of fish meat at refrigerator condition and it reduced the total volatile basic nitrogen in the fish meat [41]. Nutmeg essential oil at 20ppm concentration has shown oxidative and microbial stability of the cooked sausage. Thiobarbituric acid-reactive substance values and total number of aerobic mesophilic bacteria were lowest in sausages containing 20ppm of nutmeg oil that confirmed its ability to extend the shelf life of the cooked sausage. However, the addition of essential oil caused no noticeable color change in the sausages [42].

Natural antioxidant and antibacterial effect of nutmeg oil (0.05%) also found its application in the preservation of chicken patties without affecting its sensory qualities. However, the study findings concluded that sensory scoring for flavor and acceptability of patties were reduced with increased concentration of nutmeg oil from 0.05% to 0.1% [43]. Nishad et al. in 2018 reported the synergistic antioxidant effect of binary mixtures containing nutmeg and citrus peel extract at concentration of 0.5% and 1.0%, respectively. Their combination has shown better oxidative stability of the mutton meat balls during its prolonged storage at frozen condition. In contrary to the above-stated studies, binary mixture at higher concentration (1.0%) was found to be effective against lipid and protein oxidation without compromising sensory qualities [44]. The natural preservative effect of M. fragrans has been explored in the gelatin-based chewable tablets. Various compositions of chewable tablets with seed extract and essential oil were prepared. The study findings have shown the protective effect of M. fragrans-contained tablets against mold growth and prolonged their shelf life 62 times higher than the gel tablets containing 0.96% citric acid solution as preservative [45]. Zhu et al. in 2020 [46] reported high preservative effect of plasma-treated nutmeg oil encapsulated solid liposomes in pork tenderloins batter. Addition of these solid liposomes helped in strengthening the protein–protein interaction and network formation in the pork meat batter. This formed network has shown an increased capacity to entrap water, therefore, retarded protein decomposition by oxidation. Along with this preservative effect, texture and color of the pork meat batter were maintained [46].

17.7.3. Therapeutic benefits

17.7.3.1. Traditional uses

Among Arabs of Israel and Jewish communities, especially Yemennities, nutmeg is used as a spice as well as traditional medicine. Generally, it is used in cold, fever, respiratory problems, and to treat tuberculosis. It is used to control bowel movements and manage vomiting. It is reported to have anthelmintic effect and used to alleviate skin diseases such as eczema and scabies [47]. In the Unani system of medicine, it is used in the management of male sexual disorders such as libido and potency [48]. In Western and Chinese herbal medicine, it is used to expel gas from the stomach and reduce problems related to indigestion. It is used as a muscle relaxant and reduces abdominal pain. In Indochina, powdered seeds are added to boiled rice to treat dysentery, anorexia, and colic. In Indonesia, mace is used to treat rheumatism and pain [49].

Moreover, nutmeg is traditionally used for inflammatory conditions such as joint pain and muscle spasm. Nutmeg oil is reported to treat illness related to nervous system such as psychosis and anxiety. In addition to these, nutmeg oil also helps to dissolve kidney stones and also used to treat infections related to kidney. It is also useful in the treatment of diarrhea, nausea, vomiting, flatulence, and bacterial infections [49,50].

Besides these broad set of traditional uses, specific therapeutic benefits of nutmeg are explored on the basis of bioactive components (Table 17.5) such as flavonoids, sterols, phenolic compounds, essential oil, terpenes, lignans, etc. Functional role of M. fragrans which include antioxidant, antimicrobial, antidiabetic, antiinflammatory, anticarcinogenic, antiobesity, antiageing, and protective effects is also reported as shown in Fig. 17.6.

Table 17.5

Therapeutic activities explored for Myristica fragrans.

S.No. | Extract/Isolated phytoconstituents | Plant parts | Study findings | Therapeutic activity | References

1.

Ethanolic extract of nutmeg containing myristicin

Nutmeg seed

1. • Nutmeg extract at 0.7mg/mL exerted larvicidal activity against Anisakis simplex after 24h

Anthelmintic

[51]

2.

Nectandrin B

Nutmeg seed

1. • Suppresses the expression of lipogenic gene and lipid accumulation in hepatocytes through inhibitory LXRα-SREBP-1c pathway via activation of AMPK signaling in HepG2 cells

Fatty liver diseases

[52]

3.

Ethanolic nutmeg extract free from safrole and myristicin

Nutmeg seed

1. • Administration of nutmeg extract for 12 weeks increased the soleus muscle weight and increased gene expression for mycogenesis.

2. • Preserved the muscle mass via IGF-AKT-mTOR pathway and inhibited autophagy activity

Mycogenesis

[53]

4.

Ethanolic nutmeg extract

Nutmeg seed

1. • Oral dose of 200mg/kg has shown reduction in gastric lesion in ethanol induced ulcer model

2. • Histopathological analysis of stomach has shown only mild erosion in the superficial epithelium as well as few infiltrated inflammatory cells

Gastroprotective

[54]

5.

Aqueous and ethanolic mace extract

Mace

1. • Aqueous and ethanolic extract was effective against H.pylori ATCC strain with a MIC value of 500μg/mL and 125–250μg/mL, respectively

2. • Inhibitory effect of ethanolic extract against LPS-induced NO release and cytotoxicity was reported at IC_50 values of 82.19μg/mL and 26.06μg/mL

3. • EC_50 value for DPPH and ABTS antioxidant assay were 13.41μg/mL and 12.44μg/mL

Antimicrobial, antioxidant, anticancer (gastric cancer)

[55]

6.

Ethyl acetate and methanol mace extract (malabaricone C, 3-(3-methyl-5-pentyl-2-furanyl)-2(E)-propenoic acid, licarin A, maceneolignan B, elemicin

Mace

1. • Malabaricone C exhibited acetylcholinesterase inhibitory activity (IC_50 2.06±0.04μg/mL) and in vitro antioxidant activity (IC_50 6.56±0.02μg/mL)

2. • 3-(3-methyl-5-pentyl-2-furanyl)-2(E)-propenoic acid exhibited α-glucosidase inhibitory activity (IC_50 50.91±0.01μg/mL)

Antioxidant, inhibitory effect of anticholiesterase and α-glucosidase suggested for treatment of Alzheimer's disease

[56]

7.

Essential oil from nutmeg identified 25 compounds; sabinene as major component

Nutmeg seed

1. • In vitro antioxidant study by DPPH assay has reported good antioxidant potential with EC_50 of 1.35±0.003mg/mL

2. • Essential oil has showed high sensitivity to B. subtilis and C. albicans

Antioxidant and antimicrobial

[57]

8.

Essential oil (sabinene, α-pinene, α-copaene)

Nutmeg seed

1. • Oil components act rapidly and reversibly on connexin gene Cx43 which abundant in the Novikoff rat hepatoma cells.

2. • It down-regulates the Cx43 expression and reduces proliferation colony formation through a communication independent mechanism

Anticancer

[58]

9.

Acetone nutmeg extract (total phenolic content of 0.6217mg/mL)

Nutmeg seed

1. • Higher inhibition zone was observed against C. albicans (15mm)

2. • Higher phenolic content reported antioxidant effect

3. • Better inhibition of cyclooxygenase-2 activity compared to aspirin

Antimicrobial, antioxidant, antiinflammatory

[59]

10.

Essential oil (identified 33 compounds and major compounds are safrole, β-phellandrene, 3-p-menthene, L-terpinen-4-ol)

Nutmeg seed

1. • Free radical scavenging assay reported 21.95μg/mL and β-carotene in linoleic acid studies reported 67.9% inhibition of linoleic acid proved its antioxidant potential

2. • Aspergillus niger and Streptococcus mutans were found to be sensitive with MIC of 1.01 and 1.72mg/mL

3. • Cytotoxicity against nauplii of Artemia salina was recorded at LC_50 value of 31.05μg/mL with a minimum hemolytic activity of 5.21% at 0.5mg/mL

Antioxidant, antimicrobial, anticancer

[60]

11.

(2E)-5(2z.4E)-hexa-2,4-dio-zyl)-2-propylcyclohexanol

Bark

1. • DPPH assay and in vitro cell line study in MCF has reported IC_50 of 99.76 and 10.75ppm respectively

Antioxidant and anticancer

[61]

12.

Ethanolic nutmeg extract fractioned with hexane, n-butanol, ethyl acetate

Nutmeg seed

1. • Highest IC_50 value of ethyl acetate fraction (21.66μg/mL) and ethanolic extract (21.83μg/mL) have significantly induced apoptosis via caspase-3 therefore inhibited cell proliferation in melanoma malignant cell line (B16–F10)

Anticancer (skin cancer)

[62]

Abbreviation: LXRα-SREBP-1c: Liver X receptor alpha-sterol regulatory element-binding protein-1c; IGF-AKT-mTOR: Insulin-like growth factor—protein kinase-mammalian target of rapamycin; DPPH: 2,2-diphenyl-1-1-picrylhydrazyl; ABTS: 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonate); EC_50: half-maximal effective concentration; LC_50: lethal dose 50%: IC_50: half-maximal inhibitory concentration; MIC: Minimum inhibitory concentration; NO: nitric oxide.

Figure 17.6
Therapeutic effect of Myristica fragrans.

17.7.3.2. Prophylactic and protective effect

The hepatoprotective effect of isolated myristicin from hexane extract of nutmeg powder was evaluated on lipopolysaccharide (LPS)/D-galactosamine-induced liver damaged murine model. Three hours prior to the induction of liver damage, a single oral dose (200mg/kg) of myristicin was administrated and after 1h of liver damage, blood and serum samples were examined. Marked inhibition in the elevated serum level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was observed, followed by the suppression of elevated serum tumor necrosis factor α (TNF-α). During agarose gel electrophoresis, DNA fragmentation was suppressed in myristicin-pretreated group. This observation confirmed the inhibition of apoptosis in liver cells, thereby provided protection of the hepatocytes [63]. Similar to hepatoprotective activity, cardioprotective effect of aqueous nutmeg extract was demonstrated by Kareem et al. in 2009 [64] using isoproterenol (ISO)-induced myocardial infarction (MI) in rat model. The rats received single per oral dose of 100mg/kg of extract for 30 days, and then MI was induced using subcutaneous injection of ISO. Biochemical analysis of blood samples and histological examination of heart tissue were done. Rats pretreated with aqueous extract have maintained normal blood glucose level and plasma lipid profile in comparison with the untreated rats. Histological examination of heart tissues has shown a reduction in heart tissue damage in pretreated mice. These findings showed the prophylactic role of nutmeg extract against the complication of MI [64].

Similar study procedure was used to demonstrate the role of nutmeg extract in the reduction of oxidative stress-induced hepatic damages in a rat model. In this study, the aqueous nutmeg extracts were administered to the rats, and later oxidative stress induction was done using ISO. Histopathological results showed retention of normal liver architecture in pretreated rats, whereas ISO- administered rats have shown massive changes in the liver section such as massive fatty changes, infiltration of lymphocytes, loss of cellular boundary, and necrosis [65].

Nectandrin B has shown hepatocyte-protective effect against oxidative stress-induced injury by tert-butyl hydroperoxide. The protective effect was exerted through the activation of nuclear factor erythroid 2-related factor, which increased the antioxidant responsive element luciferase activity. It also promoted extracellular signal-regulated kinases activation but inactivated glycogen synthase kinase-3β (GSK-3β) through adenosine monophosphate-activated protein kinase (AMPK)-mediated inhibitory phosphorylation [66].

Nutmeg extract containing myrislignan alleviated thioacetamide-induced acute liver injury in mice model through the modulation of peroxisome proliferator-activated receptor. The reduction in hepatic inflammation and recovery of increased serum transaminases confirmed the hepatoprotective effect of myrislignan [67]. Prophylactic effect of myristicin was investigated in the acetic acid-induced colitis mice model. Pretreatment of mice with myristicin significantly increased the antioxidant enzyme activity and decreased lipid peroxidation. These findings were persistently observed in colitis-induced mice. The decrease in colonic tissue damage with marked decrease in inflammatory markers confirmed the hepatoprotective role of nutmeg through its antioxidant responses as well as nuclear factor kappa B inhibition [68].

17.7.3.3. Antidiabetic activity

Hypoglycemic and antidiabetic activities of petroleum ether extract of M. fragrans (200mg/kg) in normoglycemic and alloxan-induced diabetic rats were investigated by Somani et al. in 2008 [69]. The activity of petroleum ether extract of M. fragrans was compared with glibenclamide (sulphonyl ureas) standard treatment. Improved glucose tolerance and hypoglycemic activity of both normal and diabetic-induced rats confirmed the antidiabetic potential of petroleum ether extract, and the results obtained were comparable to that of glibenclamide-treated rats. Both glibenclamide and petroleum ether extracts exerted hypoglycemia through similar mode of action which led to increased insulin secretion from pancreatic beta-cells and increased glycogen deposition in the liver [69]. Hydroalcoholic extract of M. fragrans ameliorated hyperglycemia and abnormal lipid metabolism in chlorpromazine-induced metabolic abnormalities in Swiss albino mice model. A significant reduction (P
<.01) in glucose and triglyceride level was observed with oral administration of extract at 450mg/kg for seven days [70]. In another study, six days treatment with safrole-free concentrated ethanolic extract of the nutmeg seed in type 2 diabetic rat model has shown a 40% reduction in blood glucose level and a 50% reduction in blood triglyceride level. During luciferase assay, a dose-dependent effect of the extract was confirmed, and it was found to act through peroxisome proliferator-activated receptor-α/γ. This study’s findings substantiated the potential role of extract for antidiabetic and antidyslipidemic treatment [71]. Oral administration of 100mg/kg and 200mg/kg of petroleum ether extract of nutmeg has shown decreases in blood glucose level and an increase in serum insulin level in alloxan-induced diabetic rats. Extract-treated groups have shown noticeable changes in histopathological analysis in comparison with the untreated group. During morphological examination of the pancreas, a significant increase in pancreatic islets cells and protection of β cells was observed. The β cells protection confirmed the reduction in oxidative stress, and this antioxidant effect has shown significant effects in the exploration of antidiabetic activity [72].

17.7.3.4. Antiinflammatory, antioxidant, and analgesic activity

Essential oils of nutmeg are traditionally used in inflammatory conditions such as joint pain and rheumatism. Evidence-based experimental studies were carried out by researchers to find out the specific components and their mechanism of action involved in exerting antiinflammatory activity. The mode of action of nutmeg to exert antiinflammatory activity was similar to nonsteroidal antiinflammatory drugs such as aspirin and indomethacin [73]. These drugs inhibited the biosynthesis of prostaglandins [74]. Antiinflammatory activity of myristicin, an active component in the essential oil was evaluated on polyinosinic-polycytidylic acid-activated RAW 264.7 macrophages. It was observed that myristicin has inhibited excessive production of nitric oxide, cytokines, chemokines, and growth factors of double-stranded RNA-activated macrophages [75]. Similar to this finding, in another study, ethanolic nutmeg seed extract possesses antiinflammatory potential in a dose-dependent manner and showed inhibition of nitric oxide, TNF-α, IL-6, and IL-β production in LPS-stimulated murine macrophage cell line (RAW 264.7) [76].

Zhang et al. in 2015 [77] evaluated in vitro antioxidant and antiinflammatory activity of bioactive components present in various extracts of the M. fragrans pericarp. Bioassay-guided purification of extracts were performed and yielded neolignans, phenyl propanoids, phenolic aldehyde, and triterpenoids. The isolated neolignans such as licarin A, licarin B, 3′-methoxy-licarin B, threo-Δ8′,3,4,3′,5′-7- hydroxy-tetramethoxy-8-O-4′-neolignan, erythro-Δ8′-4,7-dihydroxy-3,3′,5′-trimethoxy-8-O-4′-neolignan reported 70% to 90% lipid peroxidation inhibition and compounds such as erythro-Δ8′-7-hydroxy-3,4,3′,5′-tetramethoxy-8-O-4′-neolignan, erythro-3,4,5-trimethoxy-7-hydroxy-1′-allyl-2′,6′ dimethoxy-8-O4′-neolignan, erythro-Δ8′-7-acetoxy-3,4,3′,5′-tetramethoxy8-O-4′-neolignan, myristicin, elemicin, 2,3 dihydroxy-5-(hydroxymethyl) benzaldehyde, and cycloartenol, 3′-methoxy-licarin B, threo-Δ8′,3,4,3′,5′-7-hydroxy-tetramethoxy-8-O-4′-neolignan, erythro-Δ8′ −4,7- dihydroxy-3,3′,5′-trimethoxy-8-O-4′-neolignan inhibited both cyclooxygenase 1 and 2 enzymes (37%–49%) [77]. Neolignan such as myrislignan and machilin D isolated from the seed of M. fragrans exhibited antiinflammatory activity by suppressing the expression of inducible nitric oxide synthase mRNA [78]. Antiinflammatory and analgesic effect of nutmeg oil was tested in complete Freund's adjuvant (CFA)-injected rat model. Reduction in joint swelling, allodynia, and heat hyperanalgesia was evaluated, and immunohistochemistry staining of the skin samples suggested decrease in cyclooxygenase 2 enzyme expression, which normally increased with the injection of CFA. These observations suggested antiinflammatory activity of nutmeg oil. In this experimental study, nutmeg oil at its high dose of 20mg/kg has shown no much elevation of substance P level in the blood confirming its potential as analgesic activity [79].

17.7.3.5. Antimicrobial activity

Various extracts and essential oil of M. fragrans have shown antimicrobial activity against various gram-positive bacteria, gram-negative bacteria and fungi which include S. aureus, B. subtilis, K. pneumonia, L. monocytogenes, and Aspergillus pneumonia. Takikawa et al. in 2002 [80] reported antibacterial activity of ethanolic nutmeg seed extract against E. coli O157 strain, which was found highly sensitive to β-pinene [80]. The differences in the spectrum of activity against various microorganism are based on the functional groups (-COOH, –COOR, –SH, –NH_2) present in the antibacterial agent. Antibacterial activity of trimyristin, myristic acid, myristicin obtained from nutmeg seed extract were evaluated against both gram-positive (S. aureus, B. subtilis, and Micrococcus luteus)
and gram-negative (P. aeruginosa and E. coli) bacteria. The presence of carbonyl group in trimyristin, myristic acid, and aromatic ring with unsaturated side chain in myristicin was responsible for antibacterial activity [81]. Acetone extract of nutmeg seeds has shown antioxidant and antimicrobial activity against S. aureus and Aspergillus niger. GC-MS analysis of the acetone extract reported highest concentration of sabinene (28.61%), followed by other components such as β-pinene, α-pinene, myristicin, isoeugenol, p-cymene, carvacrol, eugenol, and β-caryophellene [82]. Each of the above-mentioned components has reported to possess antimicrobial activity.

Dorman and Deans have reported spectrum activity of each isolated antimicrobial agents and their mode of action. β-pinene and α-pinene exerted antimicrobial activity through membrane disruption. Phenolic compounds present in the M. fragrans act on the outer cell membrane of the microorganisms resulted in membrane damage and its leakage. This alteration in the membrane integrity increased influx and efflux of protons into and out of the cell membrane resulted in antimicrobial activity. The relative position of the hydroxyl group and alkyl substitution in the phenolic nucleus determines the effectiveness of each component against microorganisms. Alkyl-substituted phenol nucleus in carvacrol exhibited enhanced antimicrobial activity than p-cymene [83]. However, a combination of carvacrol and cymene exhibited synergistic antimicrobial activity by expanding the membrane and finally caused destabilization of the membrane [84]. Methanolic extract of M. fragrans was reported to have antimicrobial activity against gram-positive and gram-negative bacteria. A zone of inhibition was measured at various concentrations (25mg/mL, 50mg/mL, 75mg/mL, and 100mg/mL) of the extract, and these concentrations were found to be effective against P. aeruginosa [85].

The presence of the methoxy group in some newly synthesized compound (2-phenyl quinolones) has shown efflux pump inhibition via NorA protein in resistant strains of S. aureus [86]. Similar to this mechanism of action, crude extract and essential oil from M. fragrans seeds have reported inhibition of bacterial efflux pumps of methicillin-resistant S. aureus (MRSA). GC-MS analysis confirmed the presence of methoxy-containing compounds such as myristicin, elemicin, and methoxyeugenol in crude extract whereas essential oil contained myristicin and cis-asarone. These compounds were found to act on NorA and MepA, chromosomal coding on the efflux pump in MRSA. This inhibitory mechanism confirmed the antimicrobial activity of the crude extract and essential oil [87].

17.7.3.6. Anticancer activity

GC-MS analysis of nutmeg oil reported 38 compounds with free radical scavenging activity and cytotoxic activity. Later MTT assay of the nutmeg oil was carried out on human colorectal carcinoma (HCT-116) and human breast carcinoma (MCF-7) cell lines. The oil has reported IC_50 value of 78.61μg/mL in HCT-116 cell line and 66.45μg/mL in MCF-7 cell line, which demonstrated its potential cytotoxic activity [88]. Methanolic extract of M. fragrans induced apoptosis and inhibited cell proliferation in Jurkat (immortalized human leukemia) cell line. A dose dependent inhibition occurred through down-regulation of sirtuin 1 (SIRT 1) mRNA gene expression [89]. Similarly, methanolic extract of M. fragrans was found to be effective against the growth of human osteosarcoma MG-63 cell lines [85]. During the MTT assay, various concentrations of mace extract ranging from 25 to 125μg/mL have revealed the cytotoxic effect on human oral epidermal carcinoma KB cell lines. Further studies were carried out by western blotting technique and real-time polymerase chain reaction and found that apoptosis of the cancer cells occurred due to the reduction of Bcl-2 protein and its gene expression suggesting its potential for the prevention of oral cancer [90].

17.7.3.7. Antiobesity

In obese conditions, excess energy gets accumulated in the adipocytes of white adipose tissues. The major physiological regulator involved in fatty acid and glucose metabolism is the AMPK. The changes in AMPK regulations might cause diseases such as cardiovascular diseases and type 2 diabetes [91]. Nectandrin A, nectandrin B, and tetrahydrofuran guaiacin, are the tetrahydrofuran lignans that strongly stimulate AMPK in differentiated muscle (C2C12) cells at a concentration of 5M [92]. Administration of nectandrin B fraction to high fat-induced obese animal model has shown a protective effect against weight gain and blood glucose elevation. This study suggested the potential role of nutmeg in obesity, diabetes mellitus, and metabolic diseases. Nectandrin B activates AMPK presumably through Ca2+/calmodulin kinase II activation as well as stimulates the endothelial nitric oxide synthase in human endothelial cells in a concentration-dependent manner. Along with these stimulations, enhanced expression of estrogen metabolic receptor α–dependent phosphatidyl inositol 3-kinase (PI3K) was also reported which resulted in antiobesity activity [93].

Alcoholic seed extract of M. fragrans was found to inhibit the lipid synthesis and inflammation in nonalcoholic fatty liver disease. The activity is mediated by the down-regulation of the expression of lipid synthesis-related fatty acid synthase and sterol regulatory element-binding protein 1c (SREBP-1c). This resulted in lowering of the lipid content in the hepatocytes. The extract also inhibited free fatty acids–mediated cytokines such as interleukin-6 and TNF-α expression in hepatocytes. During in vivo studies, high-fat diet has induced nonalcoholic fatty liver disease in mice. The extract-treated obese mice group has shown reduction in body weight and also reported no influence with food intake during treatment. Further, analysis revealed the reduction in inflammatory mediators’ level, cholesterol, lipid accumulation, blood glucose level, and other liver function tests that confirmed the ability of extract to down-regulate the lipid-gene expression in ameliorating nonalcoholic fatty liver disease [94].

17.7.3.8. Antiageing

Nectandrin B, a lignan specifically belongs to tetrahydrofuran guaiacin was isolated from the nutmeg seeds, which ameliorated cellular senescence in old human diploid fibroblast cell. This study suggested its role to extend the life span and age-related conditions. Nectandrin B induced protection through the activation of AMPK and mediated reactive oxygen species (ROS) scavenging activity, therefore decreased intracellular ROS level. It also reduced the activity of β-galactosidase and related senescence markers which include Ser^15 phosphorylated P53, caveolin-1, P21waf1, P16ink4a, P27kip1, and cyclin D1. A noticeable extension in S-phase of the cell cycle was observed as well as a reversal effect was reported in the activity of sirtuins (SIRT1-5) and other cell survival/death-related proteins [95].

17.8. Focus on future perspective of nutmeg

Researchers have worked a lot on identification and isolation of several phytoconstituents, and later several activity-based studies have been carried out. Nowadays, the focus is toward formulation development to evaluate the effectiveness of the formulation using in vitro or in vivo model. Some of the recent research work related to formulation development is discussed in this section.

Powdered nutmeg filler in polyurethane composite foam was found to be effective against gram-negative (E. coli) and gram-positive (S. aureus) bacteria [96]. The effectiveness of nutmeg oil against
S. aureus resulted in the development of nutmeg seed oil (10%) microemulsion for acne treatment [97]. Mohd et al. utilized high-pressure homogenization for the development of nutmeg oil–loaded nanoemulsion containing surfactant, glycerol, and water. The developed formulation was found to be effective against Aedes aegypti. The controlled release of the oil from nanoemulsion has provided prolonged mosquito repellant activity [98]. The larvicidal activity of cellulose nanocrystal-stabilized Pickering emulsion of nutmeg essential oil at 50μg/mL was found to be 100% effective against Aedes albopictus [99]. Nutmeg oil (25%) containing antiseptic gel was formulated to prevent the spread of antibacterial agent [100]. In vitro antidiabetic activity of silver nanoparticle containing hydroethanolic extract of M. fragrans seed has shown inhibitory effect on α-amylase and α-glucosidase activity. Further performed, glucose diffusion and uptake assay, which showed that the silver nanoparticles with extract have retarded the glucose transport across the membrane. This proves the therapeutic efficacy of the developed formulation for the treatment of diabetic mellitus [101].

In food industry, nutmeg powder, oleoresin, or oil are incorporated into food stuff to improve the physical, textural, and functional attributes of the food stuffs or to enhance shelf life of the product vis a vis antioxidant activity. The addition of 1% microcapsules of spray-dried nutmeg oleoresin with 1:1 ratio of gum Arabic and sorghum starch into donut formulation has improved the physical, textural, and functional attributes of deep-fried donuts. These improvements are contributed by the antioxidant activity of the higher phenolic content and flavonoid content of the total mixture [102].

17.9. Toxicity aspects of M. fragrans

Development of nutraceutical and pharmaceutical formulations of M. fragrans seed is limited due to the reported oral toxicity of myristicin and elemicin. It has reported toxicological effect on the nervous system (drowsiness, numbness, delirium, and paresthesia), digestive system (vomiting and ileus), and cardiovascular system (tachycardia and hypotension). Five grams of nutmeg powder contain 1 to 2mg of myristicin, which caused psychogenic effect, and above this dose, it can be lethal to cause death [103]. Rahman et al. in 2015 reported the hallucinogenic effect of M. fragrans seed containing myristicin [104]. After consumption of nutmeg, myristicin undergoes metabolic process, which resulted in the formation of 3-methoxy-4,5 methylenedioxyamphetamine, which belongs to amphetamine class of drug but have activity similar to lysergic acid diethylamide [105]. Recently dose- and duration-dependent hepatotoxic effect of M. fragrans seed was reported. The study was carried out in male mice with a low dose (1.0g/kg) and high dose (4.0g/kg) of nutmeg powder. The selected doses were administered in normal saline by intragastrical gavage to each mouse for 7 and 14 days. Both low and high dose-treated mice livers were examined histologically after 7 and 14 days. The dilation of the central hepatic vein of the hepatic lobules and venous lumen of the portal area were observed. Also, lymphocyte infiltration and hepatocyte necrosis were reported slightly in low dose treatment for seven days. However, higher dilation in hepatocytes, inflammatory infiltration, and necrosis were observed with low dose for 14 days and high dose for seven days. Routine liver function tests are based on serum AST and ALT level. Exposure of low- and high-dose level of nutmeg has shown significantly elevated level of AST and ALT. These findings therefore confirmed the toxic effect of nutmeg powder on hepatocytes. Moreover, metabolomics analysis of liver tissues was done to investigate the differential metabolites and their pathways involved in nutmeg exposure. Several differential metabolites were found to be involved in the up-regulation and down-regulation of the different metabolic pathways and resulted in hepatotoxic effects [106].

17.10. Conclusion

M. fragrans has wide acceptability among the population for its medicinal and therapeutic uses. Its traditional application for the treatment of various illnesses and ailments has resulted in extensive research with its whole extract, isolated fractions, and essential oils. The bioactive compounds in the nutmeg have contributed toward more therapeutic uses such as antioxidant, antimicrobial, antiinflammatory, protective effects, and anticancer activities. Even though it has been known for therapeutic applications, but its use as a spice and flavoring agent in cuisine is more popular. The volatile oil present in it provides antimicrobial and antioxidant activity; hence, it would be used as natural food preservative. When volatile oils are added into food stuff, it causes no loss of organoleptic properties and retard microbial contamination too, thereby reduces foodstuff spoilage. Hence, food industries give priorities for the preservation of food stuff with natural component, which combat free radical-mediated organoleptic deterioration and improve the shelf life of the products. The nutritive and pharmacological activities of bioactive components present in spices are utilized; however, the toxicity profile of them needs to be explored, before considering them for future clinical developments.

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Chapter 18: “Ashtawarga”: the wonder medicinal herbs of nature

Vikas Gupta
[1]
, Gunpreet Kaur
[1]
, Mukesh Maithani
[2]
, Renu Bansal
[3]
, and Parveen Bansal
[1]

1
University Centre of Excellence in Research, BFUHS, Faridkot, Punjab, India
2
MRU, VCSG Government Institute of Medical Sciences & Research, Srinagar, Uttarakhand, India
3
Government Medical College, Patiala, Punjab, India

Abstract

Due to role of nutraceuticals in health care, herbal products have become major concern for food industry and health-care regulatory bodies. The World Health Organization estimates that Ayurveda is gaining popularity due to its easy availability, low cost, congeniality, and higher safety. Ashtawarga group of plants used in a number of aphrodisiacs Ayurvedic formulations/functional foods have been included in the Red List of Threatened Species. To overcome this problem, permission to use official substitutes is being exploited by manufacturers resulting in serious problem of adulteration with spoiled, inferior, and spurious drugs. Plant dealers have discovered the “scientific” methods to adulterate a high-quality material, which is difficult to analyze without high-end equipment and chemical markers. Confirmation of text-based use of these plants may further throw light on medicinal and therapeutic potentials of these eight plants and could be a great step toward high-quality nutraceuticals in future.

Graphical abstract

Keywords

Adulterants; Ashtawarga plants; Herbal products; Nutraceutical; Substitution

18.1. Introduction

India is bestowed by nature with the wealth of fascinating and alluring medicinal plants from times immemorial. Oldest text repositories Rig Veda and Atharva Veda have clearly recognized plants and plant products in the treatment of diseases [48]. Ayurveda principles were not present in text form; however, these were practiced from generations only on mouth-to-mouth basis. This tradition passed on from several generations till the Ayurveda was compiled and found its rightful place at the end of the fourth Veda known as Atharva Veda. Ayurveda faced a downgrade due to a varied vision of rulers at different times. This caused the upliftment and introduction of other therapies including Unani and Tibbi, and volumes of good Ayurvedic literature was lost. This finally led to a decline in the glory of Indian medicine resulting in loss of number of existing effective remedies. It gave rise to a number of worthless drugs of doubtful origin lacking curative properties, and thus the good name of the Indian system of medicine got overshadowed [49]. The same happened with Ashtawarga group of medicines too. “Ashtawarga” is a group of eight drugs (Jivaka, Rishbhaka, Meda, Mahameda, Kakoli, Kshirakakoli, Riddhi, and Vriddhi) as listed in Table 18.1 that form an important component of a number of Ayurvedic preparations including Chywanprash [67]. Despite of a number of therapeutic potentials attributed to Ashtawarga group, till yet no protocols could be designed even for proper identification of all eight plants. Due to the lack of such scientific literature, a setback has been faced by such potential drugs. There is a strong need to develop scientific literature on these plants, and efforts have also been made by the authors to develop monographs on existing scientific studies related to that all sorts of plants are indiscriminately used as “Ashtawarga” group of plants.

18.2. Home of Ashtawarga

The Himalayas is a great reservoir of natural vegetation important from medicinal and nutritional point of view. As per estimates, about 35,000 to 70,000 plant species are used in the preparation of medicinal products worldwide [37,62,94]. According to a recent data from the last decade, about 1600 species of plants have been traditionally used in India for medicinal purposes [77]. The Himalayas is recognized as one of the storehouses for 8000 species of flowering plants [76,84,85]. Each one of these plants exists in a specific territory in the Himalayas, particularly the North-West. In addition to north-west Himalayas, some members of Ashtawarga have been reported to be occurring in adjoining Asian countries as shown in Fig. 18.1.

Table 18.1

Eight herbs of Ashtawarga group and part used for therapeutic purposes.

Common names | Botanical name/Family | Plant part
Kakoli | Roscoea purpurea wall/Zingiberaceae | Root
Kshirakakoli | Fritillaria roylei hook/Liliaceae | Root
Jeevaka | Microstylis muscifera ridley/Orchidaceae | Bulb
Rishbhaka | Malaxis acuminata D. Don/Orchidaceae | Pseudobulb
Meda | Polygonatum cirrhifolium (Wall.) royle/Liliaceae | Rhizome
Mahameda | Polygonatum verticillatum (L.) All/Liliaceae | Rhizome
Riddhi | Habenaria edgeworthii H.f./Zingiberaceae | Root
Vriddhi | Habenaria intermedia D. Don/Orchidaceae | Root

18.3. Lack of standardization of Ashtawarga plants

Keeping in view the varied benefits of Ashtawarga plants, the demand is not being compensated by market supplies of genuine drug. Red List of Threatened Species compiled by the International Union for Conservation of Nature (IUCN) and Natural Resources has included 560 plants in India, including 247 species are in the threatened category requiring immediate conservation strategies for protection. Ashtawarga plants also fall under this category due to indiscriminate use in Ayurvedic formulations and limited distribution in their natural habitats [14,26,51,90]. Majority of the Ashtawarga plants are facing different degrees of threats to their survival due to enumerable number factors like genetic drift and anthropogenic pressures, habitat loss, the fragmentation of populations, indiscriminate collections, and other human interventions (Table 18.2).

Commercial exploitation of orchids has been banned to protect these species in their natural habitats by the Government of India. Nowadays, due to rising demands and lack of adequate supply, substitutes of Ashtawarga plants are commonly used. Ministry of AYUSH has suggested the use of substitutes as shown in Table 18.3. However, provision of these substitutes has given a long rope to manufacturers to use spurious low-quality adulterants [9,54,55]. A survey reveals that about more than 60% pharmacological actions and Ayurvedic attributes of Ashtawarga plants do not match with their substitutes that may cause reduced efficacy final formulations giving a bad name to the pathy [26,51,90]. The manufacturers are exploiting the situation only because the regulatory bodies don't have stringent tools (marker compound) needed for identification/authentication of authentic plant. Till date, none of the Ashtawarga group of plants is supported by recognized chemical markers that could identify the authentic plant. This lack of information gives much liberty to herbal drugs manufacturers to use substitutes/cheap adulterants.

Figure 18.1
Distribution of Ashtawarga plants Himalayan region along with adjoining Asian counties.

Table 18.2

Status of Ashtawarga group of plants.

S. No. | Ashtawarga plants | Status of Ashtwarga group of plants as per red list
Nearly threatened | Threatened | Endangered | Critically endangered species | Vulnerable | Extinct
1. | Kakoli | | √ | | | |
2. | Kshirakakoli | | | | √ | |
3. | Jeevaka | | | | √ | |
4. | Rishbhaka | √ | √ | | √ | |
5. | Meda | | √ | | | |
6. | Mahameda | √ | | √ | | √ |
7. | Vriddhi | | √ | √ | | |
8. | Riddhi | | | √ | | |

Table 18.3

Substitutes of Ashtawarga plants recommended by AYUSH.

S. No. | Biological source (common name) | Substitutes | Single substitute for pair of plants
1. | Roscoea purpurea (Kakoli) | Withania somnifera (Aswagandha) and Curculigo orchioides (Kali musali) | Ashwagandha
2. | Fritillaria roylei (Kshirakakoli) | Withania somnifera (Aswagandha) and Chlorophytum arundinaceum (Safed musali).
3. | Microstylis muscifera (Jeevaka) | Pueraria tuberose (Vidarikand) and Centaurium roxburghii (Lal behmen) | Vidarikand
4. | Malaxis acuminata (Rishbhaka) | Pueraria tuberose (Vidarikand), Centaurea behen (Safed behmen), and Tinospora cordifolia (Guduchi)

5.

Polygonatum cirrhifolium (Meda)

Asparagus racemosus (Satavari), Sida veronicifolia (Nagbala), Polygonatum multiflorum (Shakakul mishri), and Paederia foetida (Prasarani).

Shatavari

6. | Polygonatum verticillatum (Mahameda) | Asparagus racemosus (Satavari) and Eulophia campestris (Salam mishri).
7. | Habenaria intermedia (Vriddhi) | Sida cordifolia, Bala (Varahikand), and Asparagus filicinus (Chiriya musali) | Varahikand

8.

Habenaria edgeworthii (Riddhi)

Tacca integrifolia (Varahikand), Dactylorhiza hatagirea (Salam panja), and Habenaria griffithii/Sida acuta Burm (Mahabala)

18.3.1. Steps taken toward standardization

A very low volume of work has been done to standardize these plants with respect to modern parameters of quality control. In India, there are very few laboratories outfitted with the knowledge and expertise that are required for quality control of medicinal plants. The problem of poor-quality assessment of herbal drugs may be due to the lack of certain intrinsic factors such as active principles, testing procedure for measuring their quantities, nonavailability of the active/marker compounds apart from the phytochemical variability of plant material in relation to geography, season, age, method of harvest, drying process, etc. It is well known that the quality of the finished product is directly proportional to the quality of the raw material used. Identification of plant material through microscopic and other chemical methods will not provide absolute information on the adequacy of the material. In order to ensure the quality, safety, and efficacy of herbal products/formulations, it is very much essential to estimate the active principle or the biological active marker compound by using suitable chromatographic techniques such as chromatographic fingerprinting, column chromatography, thin-layer chromatography, high-performance thin-layer chromatography, etc. Systematic investigations using chemical markers may lead to discoveries and the development of new drugs. Our research team has isolated 14 chemical marker compounds from the Ashtawarga group of plants as shown in Table 18.4 by using chromatographic techniques.

Table 18.4

Chemical marker compounds isolated from Ashtawarga plants—First Report.

S. No. | Name of the plant | Plant part | Isolated compounds | References
1. | Jeevaka (Microstylismuscifera) | Bulb | N-isopropyl-palmitamide | [65,69]
5-O-Caffeoylquinic Acid | [65,69]
Sitostanol Caffeate (stigmastanol Caffeate) | [66]
2. | Kakoli (Roscoeapurpurea) | Root | Lupenone | [16]
Epigallocatechin | [17]
Sitostanol Caffeate (stigmastanol Caffeate) | [18]
Epicatechin | [24]
3. | Kshirakakoli (Fritillariaroylei) | Root | Sitostanol Caffeate (stigmastanol Caffeate) | [35]
Stigmasterol | [39]
4. | Mahameda (Polygonatum verticillatum) | Rhizome | Ferulic acid | [115]
Quinine | [115]
Moupinamide | –
5. | Meda (Polygonatum cirrhifolium) | Rhizome | Maleamic acid | [113]
6. | Riddhi (Habenariaedgeworthii) | Root | 2, 4–Dimethoxy–10–Methyl Chromeno [2,3-b] indole | –
7. | Vriddhi (Habenaria intermedia) | Root | Sinapic acid | [112]
8. | Rishbhaka (Malaxis acuminata) | Pseudobulb | – |

18.3.2. Scientific basis of pairing between Ashtawarga plants

Ministry of AYUSH allowed the use of Pratinidhi dravyas, also known as the official substitutes in the preparation of formulations in absence of authentic plants [2,91,92]. Ancient text book Bhavaprakasha Nighantu by Bhavamishra has also advised the use of the substitutes of Ashtawarga plants and classified them into four pairs Jeevaka-Rishbhaka, Meda-Mahameda, Kakoli-Kshirakakoli, and Riddhi-Vriddhi [8]. Even the plants in the pairs have been suggested to be used as substitutes for each other, e.g., Riddhi can be substituted with Vriddhi and vice versa [80]. Similar is the case of plants in other pairs of Ashtawarga plants. There must be some reason for pairing of these plants; however, no text has explained the scientific basis of the pairing [56,61]. It is likely that different Ashtawarga plants in different pairs might be complementing or supplementing some therapeutic/pharmacological actions in the body. Hence it was of great importance to find out the scientific logic behind this pairing. It was found that, the pairing between Ashtawarga plants was done on the basis of their Rasa panchakas. The plants having similar Rasa panchakas are paired together. It has been found that Rasa panchakas are interrelated to each other and it includes Rasa, Guna, Virya, Vipaka, and Prabhava as shown in Table 18.5. Charaka stated that a drug is like a fabric material, and its properties and action are like the yarn of the fabric. As per Ayurveda, physiological and biochemical effects of drugs and their mechanism of action in the body can be illustrated by pharmacodynamic principles namely Pancha mahabhutas (penta elements), i.e., Rasa, Guna, Virya, Vipaka, and Prabhava (Table 18.6), in terms of bodily components such as Tridosha (Vata, Pitta, and Kapha), Sapta dhatu (different tissues), mala (morbid factors including urine, feces, etc.), srotas (channels both macrovessels and microvessels), and agni (enzymes andand hormones) [15].

As per Ayurvedic principles, Pratinidhi dravyas can be used on the basis of their Guna-Karma properties, whereas substitution of Ashtawarga plants do not seem to follow any such Ayurvedic principles except some similarities in Rasa, Guna, Virya, Vipaka, and Doshic action (Table 18.7). This is an era of evidence-based medicine, and the most important criterion for the selection of Pratinidhi drugs is pharmacological actions rather than similarities in Ayurvedic parameters. In case of defining substitutes for Ashtawarga plants, neither such pharmacological profile nor any Ayurvedic principles have been taken care of. This may spoil/deteriorate/downgrade the quality and efficacy of such formulations that could result into consumption of undesired drug materials and may finally create a bad impression toward efficacy and sanctity of Ashtawarga-based Ayurvedic/traditional medicine and the system on the whole (Virk et al., 2015).

Table 18.5

Scientific basis of existence of Ashtawarga plants in pairs as per Ayurveda and pharmacological parameters.

S. No. | Pairs of Ashtawarga plants | Similarity in
Rasa | Guna | Virya | Vipaka | Doshic action | Pharmacological action
1. | Kakoli and Kshirakakoli | √ | × | √ | √ | – | √
2. | Jeevaka and Rishbhaka | √ | × | √ | √ | √ | √
3. | Meda and Mahameda | √ | × | √ | √ | √ | √
4. | Vriddhi and Riddhi | √ | √ | √ | √ | √ | √

Table 18.6

Effects of drugs and their mechanism of action as per Pancha mahabhutas.

Rasa panchakas | Description | References

Rasa

It is the unique intelligence of the tongue or taste buds (Rasana or Rasanendriya) or is something experienced by an individual while consuming a drug and has a direct influence on Vata, Pitta, and Kapha. Rasa plays an important role in restoring the balance of the Doshas.

[114]

Guna

It is a physical property, which will remain in a drug with inherent relationship. Gunas can be detected through their action. Certain Gunas on contact with tongue are subjected to initial digestion and exert certain actions that are attributed to Rasa.

[86], [114]

Virya

It is considered as the most potent quality of a drug/active principle or a factor, which is responsible for drug action/pharmacological action. Virya refers to potency felt immediately or some time later after tasting a drug.

[114]

Vipaka

Vipaka is defined as the last stage of transformation of drug after digestion/metabolism through the action of Jataragni (digestive enzymes). Vipaka is of three types, i.e., Madhura (sweet-deeply nutritive and building), Amla (sour enhances digestive fire), and Katu (pungent increases elimination).

[114]

Prabhava

The peculiar action of a drug is known as Prabhava and is characterized by specific actions of substances that cannot be explained in terms of the pharmacological actions of various constituents of drug considered individually in relation to each other.

[96], [114]

Table 18.7

Ashtawarga plants and their substitutes.

Description | Ashtawarga plant | Name of Substitutes
Common name | Mahameda | Shatavari | Prasarani | Nagabala
Botanical name | Polygonatum cirrhifolium (Wall.) Royle | Asparagus racemosus willd. | Paederia foetida linn. | Sida himilis Car.
Rasa | Madhura | Madhura, Tikta | Tikta | Madhhura
Guna | Guru, Snigdha | Guru, Snigdha | Guru, Sara | Guru, Snigdha
Virya | Sheeta | Sheeta | Ushna | Sheeta
Vipaka | Madhura | Madhura | Katu | Madhura
Doshic action | Vata Pitta pacifying | Vata Pitta pacifying | Vata Pitta pacifying | –
Common name | Meda | Shatavari | Salam mishri |
Botanical name | Polygonatum verticillatum (linn.) allioni | Asparagus racemosus willd. | Eulophia campestris Wall. |
Rasa | Madhura | Madhura, Tikta | – |
Guna | Guru | Guru, Snigdha | – |
Virya | Sheeta | Sheeta | – |
Vipaka | Madhura | Madhura | – |
Doshic action | Vata pitta pacifying | Vata Pitta pacifying | – |
Common name | Riddhi | Varahi kand | Bala | Chiriya musli
Botanical name | Habenaria intermedia D. Don | Dioscorea bulbifera linn. | Sida cordifolia linn. | Asparagus filicinus
Rasa | Madhura | Madhura, Katu, Tikta | Madhura | Madhura
Guna | Guru, picchila, snigdha | Laghu | Guru, Snigdha, Picchila | Guru, Snigdha
Virya | Sheeta | Ushna | Sheeta | Sheeta
Vipaka | Madhura | Katu | Madhura | Madhura
Doshic action | Tridoshahar piitahar | Vata Pitta pacifying | Vata Pitta pacifying | Vata Pitta pacifying
Common name | Vriddhi | Varahikand | Salam panja | Maha bala
Botanical name | Habenaria edgeworthii hook. f.exCollet | Dioscorea bulbifera linn. | Dactylorhiza hatagirea (D. Don) Soo | Sida rhombifolia linn.
Rasa | Madhura | Madhura, Katu, Tikta | – | Madhura
Guna | Guru, Snigdha | Laghu | – | Guru, Snigdha, Picchila
Virya | Sheeta | Ushna | – | Sheeta
Vipaka | Madhura | Katu | – | Madhura
Doshic action | Tridoshahar Piitahar | Vata Pitta pacifying | – | –
Table Continued

Description | Ashtawarga plant | Name of Substitutes
Common name | Mahameda | Shatavari | Prasarani | Nagabala
Common name | Kakoli | Ashwagandha | Kali musli |
Botanical name | | Withania somnifera Dunal. | Curculigo orchioides Gaertn. |
Rasa | Madhura | Tikta, Kashaya | Madhura, Tikta |
Guna | Sheeta, Guru | Laghu | Guru, Picchila |
Virya | Sheeta | Ushna | Ushna |
Vipaka | Madhura | Madhura | Madhura |
Doshic action | Pitta-Vatahar Kaphakar | Vata Pitta pacifying | Vata Pitta pacifying—Kapha aggravating |
Common name | Kshirakakoli | Ashwagandha | Safed musli |
Botanical name | Fritillaria roylei hook. | Withania somnifera Dunal. | C hlorophytum horivilianum F. |
Rasa | Madhura | Tikta, Kashaya | Madhura, Tikta |
Guna | Guru, Snigdha | Laghu | Snigdha, Picchila, Guru |
Virya | Sheeta | Ushna | Ushna |
Vipaka | Madhura | Madhura | Madhura |
Doshic action | Vata Pitta pacifying | Vata Pitta pacifying | – |
Common name | Rishbhaka | Vidarikand | Lal behman |
Botanical name | Microstylis muscifera (lindl.) ridl. | Pueraria tuberose DC. | Centaurium roxburghii (D. Don) Druce. |
Rasa | Madhura | Madhura | – |
Guna | Guru, Snigdha | Guru, Snigdha | – |
Virya | Sheeta | Sheeta | – |
Vipaka | Madhura | Madhura | – |
Doshic action | Vata Pitta pacifying | Vata Pitta pacifying | – |
Common name | Jeevaka | Vidarikand | Safed behman | Giloe
Botanical name | Malaxis acuminata D. Don. syn. Microstylis wallichii lindl. | Pueraria tuberosa DC. | Centaurea behen linn. | Tinospora cordifolia (willd.) miers.
Rasa | Madhura | Madhura | – | Tikta, Kashaya
Guna | Picchila, Snigdha | Guru, Snigdha | – | Laghu
Virya | Sheeta | Sheeta | – | Ushna
Vipaka | Madhura | Madhura | – | Madhura
Doshic action | Vata Pitta pacifying | Vata Pitta pacifying | – | Tridoshic

18.4. Uses of Ashtawarga plants in traditional and modern system of medicine

Ashtawarga plants have been useful for the treatment of sexual disorders (65%), physical weakness (21%), immunity imbalance (8%), and body pain (6%) (Fig. 18.2) [10]. There has been a rapid decline in fertility in male and rapid increase in reproductive disorders. As per WHO data, a 50% decrease in reproductive capacity was reported in infertile couples. Today, even after 30 years, according to a recent report, it is apprehended that Japan and the European Union will soon record an appreciable decline in their populations due to persistently low total fertility rates [87]. A number of endocrine/congenital/immunological/iatrogenic factors may decrease conception rate in the females, whereas ejaculatory dysfunction, oligozoospermia, and sexual dysfunction could be main causes in men [106]. Despite the availability of a number of synthetic drug modules which are expensive and provoke serious adverse effects, the herbal products are very much in demand [6]. While drugs like Viagra enable the patient's system to respond, whereas the Ayurvedic medicines based on traditional knowledge use plants and other natural resources to nourish and rebuild body tissue as shown in Table 18.8. In Ayurveda, this part of human behavior had been attributed to the presence of Rasayana and Vajikarna drugs (Roscoea procera, Microstylis wallichii, Mucuna pruriens, Asparagus racemosus, and Polygonatum verticillatum) that increase sexual pleasure, libido, potency, and remove the disorders of sexual desire, ejaculation, orgasm, and erectile dysfunction drugs, which increase the quantity of semen or stimulate the production of semen [4]. In Charaka Samhita, this group of eight plants along with other plants has been described to exhibit spermatogenic activities [81].

In the modern system of medicine, Ashtawarga plants showed number of pharmacological activities such as antioxidant, antianxiety, cytotoxic, antiinflammatory, antifungal, antimalarial, antinociceptive, etc., well established by different scientists are listed in Table 18.9. The analysis of literature reveals that formulations containing Ashtawarga plants are used by trado-medical practitioners (TMPs), and no systematic clinical trial data/clinical reports or such studies exist in literature that could prove to be useful in highlighting the real therapeutic potentials of these formulations. Table 18.10 showed the lack of clinical studies/clinical trials of formulations contacting Ashtawarga plants as per the modern system of medicine for use indicated as per textual uses and TMPs. This template may help the scientists/researchers for further screening on these formulations that are useful in plethora of disorders.

Figure 18.2
Use (in %) of Ashtawarga plants in traditional system of medicine.

Table 18.8

Various Ashtawarga plants in important Ayurvedic formulations useful in reproductive disorders.

S. No. | Formulation and its uses | Excerpts from Ancient textual Charak Samhita | Name of Ashtawarga plant used | References

1.

Vrishya pooplika (potent aphrodisiac)

फलानां जीवनीयानो स्त्रिग्धान रूचिकारिणाम् ।

कुडवश्चूर्णितानां स्यात् स्वयड्गुमाफलस्य च॥१५॥

कुडवश्चैव माषाणां द्वौ च तिलमुद्र्यो:।

गोघूमशालिचूर्णानां कुडव: कुडवो भवेत्॥१६॥

सर्पिप: कुडवश्चैकरूतत् सवै क्षीरमर्दितम्।

पक्तवा पूपलिका: खादेद्वहृय: स्युर्यस्व योषित॥१७॥

Kakoli, kshirakakoli, Jeevaka, rishbhaka, meda, mahameda, vriddhi, riddhi

[32]

2.

Apatyakari shashtikadi Gutika (promotes male fertility)

आसिक्त क्षीरमापूर्णमशुष्कं शुद्ध षष्टिकम्|
उदूखले समापोथ्य पीडयेत् क्षीरमर्दितम्||३||
गृहीत्वा तं रसं पूतं गव्येन पयसा सह|
बीजानामात्मगुप्ताया धान्य माष रसेन च||४||
बलायाः शूर्पपर्ण्योश्च जीवन्त्या जीवकस्य च|
ऋद्ध्यर्षभक काकोली श्वदंष्ट्रा मधुकस्य च||५||
शतावर्या विदार्याश्च द्राक्षा खर्जूरयोरपि|

संयुक्तं मात्रया वैद्यः साधयेत्तत्र चावपेत्||६||

Kakoli, kshirakakoli, Jeevaka, rishbhaka, meda, mahameda, vriddhi, riddhi

[27]

3.

Jeevaneeya Gana (enlivening, antiaging group of herbs)

तद्यथा-जीवकर्षभकौ मेदा महामेदा
काकोली क्षीरकाकोली मुदपर्णीमाषपर्ण्यो
जीवन्ती मधुकमिति दशेमानि जीवनीयानि भवन्तिII

Jeevaka, rishbhaka, meda, mahameda, kakoli, kshirakakoli,

[28]

4.

ShukralaShukrajanana (improves quality of semen and ovum)

जीवकर्षभक काकोली क्षीरकाकोली

मुदपर्णी माषपर्णी मेदावृद्धरूहा जटिला कुलिड्गा

इति दशेमानि शुक्रजननानि भवन्तिII

Jeevaka, rishbhaka, kakoli, kshirakakoli, meda,

[28]

5.

Snehopaga (adjuvants of snehana/oleation therapy in various disorders)

मृद्वीका मधुक मधुपर्णी मेदाविदारी काकोली क्षीरकाकोलीII

जीवक जीवन्ती शालपर्ण्य इति दशेमानि स्नेहोपगानि भवन्तिII

Meda, kakoli, kshirakakoli

[28]

6.

Shukrala (improves quality of semen and sperm)

शुक्रलैर्जीवनीयैश्च बृंहणैर्बलवर्धनैः|
क्षीरसञ्जननैश्चैव पयः सिद्धं पृथक् पृथक्||६||
युक्तं गोधूमचूर्णेन सघृतक्षौद्र शर्करम्|
पर्यायेण प्रयोक्तव्यमिच्छता शुक्रमक्षयम्||७||

Kakoli, kshirakakoli, Jeevaka, rishbhaka, meda, mahameda, vriddhi, riddhi

[28]

Table Continued

S. No. | Formulation and its uses | Excerpts from Ancient textual Charak Samhita | Name of Ashtawarga plant used | References

7.

Amruta prasha Ghruta (cures cough, hiccup, fever, asthma, burning sensation, thirst, vomiting, fainting, female genital tract, urinary tract infections, and diseases of the heart.

It helps in the procreation of a male child also.

जीवकर्षभकौ वीरां जीवन्तीं नागरं शटीम्|

चतस्रः पर्णिनीर्मेदे काकोल्यौ द्वे निदिग्धिके||३५||

पुनर्नवे द्वे मधुकमात्मगुप्तां शतावरीम्|

ऋद्धिं परूषकं भार्गीं मृद्वीकां बृहतीं तथा||३६||

शृङ्गाटकं तामलकीं पयस्यां पिप्पलीं बलाम्|
बदराक्षोट खर्जूर वातामाभिषुकाण्यपि||३७||
फलानि चैवमादीनि कल्कान् कुर्वीत कार्षिकान्|
धात्री रस विदारीक्षुच्छाग मांस रसं पयः||३८||

Jeevaka, rishbhaka, meda, mahameda, kakoli

[29]

8.

Ksheerayoga (mmedicated milk andjiggery with kakoli cures seminal and menstrual diseases, depletion of body tissue, emaciation, and phthisis.

स्थिरा सिता पृश्निपर्णी श्रावणी बृहती युगैः|
जीवकर्षभ काकोली तामलक्यृद्धि जीवकैः||१०१||
शृतं पयः पिबेत् कासी ज्वरी दाही क्षतक्षयी|

तज्जं वा साधयेत् सर्पिः सक्षीरेक्षुरसं भिषक्||१०२||

जीवकाद्यैर्मधुरकैः फलैश्चाभिषुकादिभिः|

कल्कैस्त्रिकार्षिकैः सिद्धे पूतशीते प्रदापयेत्||१०३||

Jeevaka, rishbhaka, kakoli,

[33]

9.

Mahamayura Ghrita (used for inhalation therapy for serious types of coughs, asthma, enema, and massage.

Also used in diseases of the female genital tract and helps in the procreation of a male offspring.

एतेनैव कषायेण घृत प्रस्थं विपाचयेत्|
चतुर्गुणेन पयसा कल्कैरेभिश्च कार्षिकैः||१६६||
जीवन्ती त्रिफला मेदा मृद्वीकर्धि परूषकैः|
समङ्गा चविका भार्गी काश्मरी सुरदारुभिः||१६७||
आत्मगुप्ता महामेदा ताल खर्जूर मस्तकैः|
मृणाल बिस शालूक शृङ्गी जीवक पद्मकैः||१६८||
शतावरी विदारीक्षु बृहती सारिवा युगैः|

मूर्वा श्वदंष्ट्रर्षभक शृङ्गाटक कसेरुकैः||१६९||

रास्ना स्थिरा तामलकी सूक्ष्मैला शटि पौष्करैः|

पुनर्नवा तुगाक्षीरी काकोली धन्वयासकैः||१७०||

Meda, mahameda, Jeevaka, rishbhaka, kakoli,

[30]

Table Continued

S. No. | Formulation and its uses | Excerpts from Ancient textual Charak Samhita | Name of Ashtawarga plant used | References

10.

Brihat shatavari Ghrita (it promotes virility and helps the woman to get a male progeny.

It is a curative of phthisis, ordinary jaundice, gout including other forms of arthritis and erysipelas)

शतावरीमूल तुलाश्चतस्रः सम्प्रपीडयेत्||६४||
रसेन क्षीरतुल्येन पचेत्तेन घृताढकम्|
जीवनीयैः शतावर्या मृद्वीकाभिः परूषकैः||६५||

पिष्टैः प्रियालैश्चाक्षांशैर्द्वियष्टिमधुकैर्भिषक्|

सिद्धे शीते च मधुनः पिप्पल्याश्च पलाष्टकम्||६६||

सिता दशपलोन्मिश्राल्लिह्यात् पाणितलं ततः|

योन्यसृक्शुक्रदोषघ्नं वृष्यं पुंसवनं च तत्||६७||

Kakoli, kshirakakoli, Jeevaka, rishbhaka, meda, mahameda, vriddhi, riddhi

[31]

11.

Milk boiled with herbs belonging to Jivaniya group (cure aamenorrhea—uterine disease)

मृगाजाविवराहासृग्दध्यम्ल फल सर्पिषा ||१०१||

अरजस्का पिबेत् सिद्धं जीवनीयैः पयोऽपि वा|

Kakoli, kshirakakoli, Jeevaka, rishbhaka, meda, mahameda, vriddhi, riddhi

[31]

Table 18.9

Evidences for pharmacological Activities of Ashtawarga plants.

S. No. | Pharmacological activity | Name of Ashtawarga plant | References
1. | Antioxidant | Roscoea purpurea (kakoli) | [93]
Fritillaria roylei (kshirakakoli) | [78]
Habenaria intermedia (vriddhi) | [68]
Microstylis muscifera (Jeevaka) | [21]
Polygonatum cirrhifolium (meda) | [25]

2.

Immunomodulator

Roscoea purpurea (kakoli)
Fritillaria roylei (kshirakakoli)
Habenaria intermedia (vriddhi)

[108]
[75]
[74]

3. | Hepatoprotecitve | Habenaria intermedia (vriddhi) | [22]
4. | Antianxiety activity | Habenaria intermedia (vriddhi) | [50]

5.

Cytotoxic activity

Roscoea purpurea (kakoli)
Fritillaria roylei (kshirakakoli)
Polygonatum verticillatum (mahameda)

[89]
[19]
[71,72]

6. | Galactogogue activity | Roscoea purpurea (kakoli) | [88]

7.

Reproductive disorders

Roscoea purpurea (kakoli)

Fritillaria roylei (kshirakakoli)

[23]

8.

Antiinflammatory activity

Polygonatum cirrhifolium (meda)

Polygonatum verticillatum (mahameda)

[42]

9.

Antipyretic activity

Microstylis muscifera (Jeevaka)
Fritillaria roylei (kshirakakoli)
Polygonatum verticillatum (mahameda)

[1]
[43]
[110]

10.

Antifungal

Polygonatum cirrhifolium (meda)

Polygonatum verticillatum (mahameda)

[53,101]

11. | Antimalarial | Polygonatum verticillatum (mahameda) | [44,46]
12. | Anticonvulsant | Polygonatum verticillatum (mahameda) | [42]
13. | Phytotoxicity | Polygonatum verticillatum (mahameda) | [71,72]

14.

Antimicrobial

Polygonatum verticillatum (mahameda)

Malaxis acuminata (rishbhaka)

[45,47]

[79]

15.

Aphrodisiac

Roscoea purpurea (kakoli),

Fritillaria roylei (kshirakakoli)

[23]

[38]

16. | Antinociceptive | Polygonatum verticillatum (mahameda) | [41]

Table 18.10

Lack of clinical studies/clinical trials of Ashtawarga plants containing formulations as per modern system of medicine for use indicated as per textual uses and trado-medical practitioners.

S. No. | Formulations | Therapeutic uses as per texts | Therapeutic uses as mentioned by trado-medical practitioners (TMPs) | Preclinical/clinical trials/case studies as per modern system of medicine

1.

Amruta prasha Ghruta/AmritaprashaGhrita

Cough, hiccup, fever, asthma, burning sensation, morbid thirst, ailment characterized by bleeding from different parts of the body, vomiting, fainting, female genital tract, procreation of male child.

Yes

No

Joint problems and other diseases caused by unbalanced vata andand pitta doshas.

Yes

Yes

Improves the nonspecific immunity against rheumatoid arthritis [100].

Heart diseases and urinary tract problems | No | No
Table Continued

S. No. | Formulations | Therapeutic uses as per texts | Therapeutic uses as mentioned by trado-medical practitioners (TMPs) | Preclinical/clinical trials/case studies as per modern system of medicine
2. | Jeevaneeya Gana | Enlivening, antiaging, antioxidant | Yes | No
3. | Jivantyadi Ghrita | Cure female infertility | Yes | No

Cure cataract and glaucoma

Yes

Yes

Effective in treatment and controlling of Computer vision syndrome and helpful in curing myopia [59,60]

4. | Khuddaka Taila/khuddaka padmaka tailam | Gout and burning sensation | Yes | No
5. | Madhuparnyadi Taila | Cures vatarakta, pain in limbs, affliction of the whole body, promotes strength and complexion. | Yes | No

6.

Mahamayura Ghrita/Mahanarayan Tailm

Neuromuscular disorders, inflammation, diseases of the female genital tract, menstrual disorders,

Yes

Yes

Used to treat inflammation, pain and arthritis, useful in treatment of chronic back pain, useful in treatment of female infertility, provide strength to the local soft tissues, thus play significant role in the management of cervical spondylosis or osteoarthritis of cervical spine [40,57,64,102]

Cough and asthma | Yes | No
7. | Mahasneham ghrita | Gout, serious diseases caused by the aggravated vata, arthritis | Yes | No
Table Continued

S. No. | Formulations | Therapeutic uses as per texts | Therapeutic uses as mentioned by trado-medical practitioners (TMPs) | Preclinical/clinical trials/case studies as per modern system of medicine
8. | Paste prepared of drugs belonging to Jivaniya group, cow-milk and muscle fat/Dhanwantharam Thailam | Cures pain | No | No
Gout and kyphosis (sspinal disorder) | Yes | No
9. | Phalakalyan grita/Taila Uttara Basti | Uterine tonic | Yes | No

Gynecological disorders

Yes

Yes

Useful in treatment of female infertility [64].

10. | Shukrala/Kamdev Ghrita | Improves semen and sperm production | Yes | No
11. | Siva Gutika/Shiva gritham | Anemia, cardiac diseases, epilepsy, psychotic diseases, infertility in women/men | Yes | No

Liver diseases

Yes

Yes

Used in treatment of chronic liver and highly effective in treatment of HIV/AIDS and acute deep vein thrombosis [83,97,103]

Diabetes mellitus, bronchitis and skin diseases | No | No
12. | Sneha parisheka | Stiffness, convulsion, gout, burning sensation | Yes | No

13.

Sukumara taila/Sukumara kashaya

Lower abdominal pain in women, menstrual pain, inflammation, constipation, promotes robustness in body and positive health

Yes

Yes

Helps in improving the quality of the ovum by regularizing the menstrual cycle which enhanced rate of conception and also used in treatment of complete rectal prolapse [83,97].

Table Continued

S. No. | Formulations | Therapeutic uses as per texts | Therapeutic uses as mentioned by trado-medical practitioners (TMPs) | Preclinical/clinical trials/case studies as per modern system of medicine
14. | Upatyakari shashtikadi Gutika | Male fertility | Yes | No
15. | Vrishya pooplika | Aphrodisiac | Yes | No

16.

Bala Ghrita/Taila

Gout and jaundice

Anemia

Yes

No

Uterine diseases, provides strength to children

Yes

Yes

Relieves constipation in pregnant women [82].

Helps in improving females' reproductive disorders [58]

Message contributes positive effects to health of new born child [82].

17.

Parushaka Ghrita

Menstrual pain and menorrhagia

Yes

Yes

Act as good uterine tonic by providing strength to uterus [98].

Anemia, respiratory disorder and jaundice | Yes | No

18.

ShukralaShukrajanana/Shukra-Shodhana-Janana Granules

Improves quality and quantity of semen and sperm

Yes

Yes

Provides strength and increases sperm count [20].

19. | Snehopaga gana/(adjuvants of snehana/oleation treatment) | Dry skin, neuromuscular disorders, muscular dystrophy, wasting disorders. | Yes | No

20.

Jivaniya mahakashaya

Provides strength, boost immunity, improves quality of semen, aphrodisiac, hormonal disturbances in women, nervousness, dizziness.

Yes

No

Table Continued

S. No. | Formulations | Therapeutic uses as per texts | Therapeutic uses as mentioned by trado-medical practitioners (TMPs) | Preclinical/clinical trials/case studies as per modern system of medicine
21. | Brihat shatavari Ghrita | Virility, mmale progeny, ppulmonary tuberculosis, jaundice, gout, sskin disorders. | Yes | No

Sexual disorders

Yes

Yes

Useful in treatment of female infertility [52].

22. | Shatavaryadi Ghrita/shatavari tailam | Cough, fever, flatulence, constipation | Yes | No

Colic pain, rectal bleeding and heavy periods

Yes

Yes

Useful in treatment of female infertility and other UTI disorders [52].

18.5. Red list category and criteria

Red List or Red Data Book is the most authoritative guide prepared by the IUCN to reflect the status of biological diversity. The status of Ashtawarga plants as per IUCN Red List is shown in Fig. 18.3.

Fritillaria roylei distributed in India, Afghanistan, and Pakistan is an important medicinal plant traded at local, regional, and national markets for its bulbs. The majority (80%) of the geographical distribution is in India. High demands result into the indiscriminate collection of this plant leading to whole plant uprooting by collectors during collection. This further has a severe impact on natural regeneration. This results in declining of the natural habitat. The Himalayan Region of India lost more than 80% of the wild population in the past several years giving rise to critically endangered (EN) species. The status has not changed even now [73] and the status in India is therefore considered representative of the global population.

Roscoea purpurea is native of damp and dry habitats of Nepal. It occurs in a range. It is mostly found growing in alpine grassland, terraced walls, and rock faces exposed to the full sun. It is also found in the shade of other herbaceous plants, shrubs, and trees. Populations of the species have limited distribution in their natural habitats. The population of Roscoea purpurea is continuously disappearing from its natural habitats and has been included in the red list [3] because of a number of varied factors such as indiscriminate harvesting, destruction of rare plants, habitat loss, unsuitable climatic conditions, deforestation, overexploitation, etc.

Figure 18.3
Status of Ashtawarga plants as per IUCN Red List.

Wild populations of Meda are continuously disappearing from its natural habitats. Many years ago, it has been included in the red list [5]. The threatened status of Meda has also come up due to the explosion of human density, land-use disturbances for various commercial projects, habitat specificity, narrow range of habitat distribution, invasion by nonnatives, unspecified climatic changes, livestock grazing, degradation of plant density, and genetic drift. In Uttarakhand, women of studied areas collect the food and fodder for animals from the nearby forests due to lack of identification and is a major reason of threatened status of Meda [63].

Mahameda is continuously disappearing from its natural habitats. It finds place in red list under different categories of threat and considered as EN, vulnerable, and near to threaten in different study areas [70].

Due to limited resources and increased market pressure, the collection of Malaxis acuminata (Rishbhaka) from the wild has been banned in Uttarakhand. No efforts have been recorded for encouraging commercial cultivation. There is a great need of developing sustainable harvesting protocols [99]; [104] and protocols to screen the different chemotypes of M. acuminata that could be instrumental in conservation of the plant [105].

Major threats to terrestrial orchid such as Microstylis muscifera (Jeevaka) species are also more or less same as explained in earlier plant species.

Wild population of Vriddhi is continuously disappearing from its natural habitats. IUCN has been included it in the list of threatened species as EN plant [7]. It has been categorized as EN in Himachal Pradesh (2003 and 2010), Uttarakhand (2003), and Jammu and Kashmir (2003) [109]. In Uttarakhand and Uttaranchal, Vriddhi is reported as highly EN plant and it may be due to overexploitation and habitat destruction by collection of plant [13,34]. At local level, very few studies have been carried out following the IUCN criteria. The local level threat categorization of the species has been considered as the best approach for developing appropriate strategies and management plan [111].

The major causes of EN status of Habenaria edgeworthii (Riddhi) are the overexploitation and habitat degradation. Therefore, habitat monitoring, bulk multiplication of the species, improved in vitro propagation protocols, creation of educational and awareness centers for the inhabitants on conservation, and participation of local natives for conservation management are suggested in order to regenerate their population.

18.6. Conservation actions and strategies for future prospects

- i. As the Ashtawarga plants undergo threatened condition due to the destruction of habitats and unsustainable collection practices, the regulatory authorities are required to develop steps for in situ and ex situ conservation of such plants along with formulating stringent trade practices. A number of workers have already worked on the development of protocols for the propagation of these plants through in vitro as well as traditional/conventional practices [11]. It has been summarized that domestic growing of such plant species may be successful in decreasing the intensity of pressure plant population in wild habitats [12].

- ii. Traditional knowledge system and systematic approach are the most important aspect of conservation. As per the Convention on Biological Diversity, environment can be managed through systematic conservation of http://www.scialert.net/asci/result.php?searchin=Keywords&cat=&ascicat=ALL&Submit=Search&keyword=medicinal+plant (medicinal plant) medicinal plants using good cultivation practices along with inclusion of new scientific methods accompanied by traditional practices [95]. This can be combined with http://www.scialert.net/asci/result.php?searchin=Keywords&cat=&ascicat=ALL&Submit=Search&keyword=in+vitro (in vitro) in vitro bulblet method and seed [36] for the restoration of population significantly.

- iii. Data collection about interfering factors, best seasons for growth, adaptable conditions, natural habitat marking for such plants under threat along with reproductive as well as vegetative phenology.

- iv. Formulation of byelaws and regulations for proper economic gains from cultivation to encourage good cultivation practices.

- v. Increase in earmarked cultivable land area for the development of national parks, medicinal plants conservation area networks, and sanctuaries for in situ conservation.

- vi. Development of herbal/botanical gardens in suitable climatic conditions for e x situ conservation.

- vii. Invention of agrotechnology tools for the convenience of farmers for bulk commercial cultivation.

- viii. Proper training on scientific methods of conservation, development, harvesting, management, etc., to the persons engaged in the harvesting [107].

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Further reading

[1]. Kaur J.V, Bansal P, Gupta V, Kumar S, Singh R, Rawal R.K. First report of isolation of maleamic acid from natural source Polygonatum cirrhifolium—a potential chemical marker for identification.
J. Liq. Chromatogr. Relat. Technol.
2017;40(20):1031–1036.

[2]. Kaur J.V, Gupta V, Kumar S, Singh R, Bansal P. Lack of pharmacological basis of substitution of an endangered plant group “ashtawarga”- A significant ingredient of polyherbal formulations.
Am. J. Phytomed. Clin. Ther.
2015;3(12):690–712.

[3]. Kaur J.V, Gupta V, Kumar S, Singh R, Rawal R.K, Bansal P. Isolation and quantification of chemical marker of Polygonatum verticillatum: First report.
Curr. Tradit. Med.
2016;2(3):216–224.

[4]. Kaur J.V, Gupta V, Kumar S, Singh R, Rawal R.K, Tripathi A.C, Saraf S.K, Bansal P. Isolation and characterization of quinine from Polygonatum verticillatum: a new marker approach to identify substitution and adulteration.
J. Adv. Pharm. Technol. Res.
2016;7(4):153–158.

[5]. Lekurwale P.S, Pandey K, Yadaiah P. Management of amavata wit ‘Amrita Ghrita’: a clinical study.
Ayurveda
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[6].
Charak Samhita, Vol. II
(Chapter 2), Qtr 1, Section 6. Varanasi, India: Chikitsasthanam. Chaukhambha Orientalia; 1998:37.

[7]. Bladt S.
Plant Drug Analysis: A Thin Layer Chromatography Atlas
. Springer Science & Business Media; 2009.

Chapter 19: Regulatory aspects of nutraceuticals

Karthik Varma A.C., Sasikumar Kuttappan, and Augustine Amalraj
R&D Centre, Aurea Biolabs (P) Ltd, Cochin, Kerala, India

Abstract

Nutraceutical supplements have always gained the accolades when it was launched into the market on account of their diverse range of pharmacological activities and clinical backgrounds. The country-wise regulations covering the manufacturing practices, use of ingredients, serving dosages, etc., can help to create an awareness among the manufacturers to set up products that meet global standards. The chapter tries to cover the regulatory aspects of nutraceuticals/food supplements in the United States, Association of South East Asian Nation, Japan, China, Australia/New Zealand, and India and the requirements of the regulatory bodies.

Keywords

FDA; Food supplements; FSSAI; Nutraceuticals; Regulations

19.1. Introduction

The use of natural supplements as an alternative to many lifestyle disorders has led researchers to develop phytochemical extracts and based formulations for treating illnesses. This concept has directed to the development of dietary supplements, which are classified as nutraceuticals to treat health disorders. The term “nutraceuticals” was introduced by Dr. Stephen DeFelice and defined as “food or part of a food that provides health or medical benefits, together with the prevention and/or treatment of a disease” [1,2]. Even though this definition was coined by scientific communities, the definition was not approved by any regulatory agency. In order to have more insights into the concept of nutraceuticals, people used to link the definition of nutraceuticals with Hippocratic principle “let food be thy medicine, and medicine be thy food,” which have directed to a misinterpretation that food and medicine are the most similar.

From the time of introduction of the concept, there was a huge demand for nutraceutical supplements on a global scenario. Dietary supplements and herbal/natural products are the two primary areas of the nutraceutical industry. Most of the nutraceuticals have substantiated the clinical efficacy with either preclinical or clinical studies [3]. Dietary supplements (including botanicals), functional foods, and medical foods are different classes of nutraceutical supplements [4]. The wide use of nutraceutical supplements has paved the way for two major requirements in the field viz: (1) individualization of their use and (2) setting of a global standard for the quality of use. The trend for nutraceutical supplements for treating lifestyle disorders have also led to the formulation of nutraceutical supplements. Diverse marketed variants can also help inhibition of lifestyle disorders into fatal diseases. Even though there is an increasing demand of nutraceutical supplements, there are no globally accepted regulatory criteria for ensuring the supplementation of the products. Country-wise regulations have been in practice to ensure the quality of goods coming to their specific nations.

The chapter tries to present an overview on various global regulatory bodies, which has the authority to implement the rules regarding the marketing and manufacturing practices of supplements.

19.2. Need for regulatory standards

Most of the countries do not regulate dietary supplements/nutraceuticals as a stand-alone category, instead, they divide it as a subcategory in the existing regulation [5]. The need for global regulatory standards emphasizes its focus on major three reasons, which include:

- 1. Promising legal security and more predictable environment.

- 2. Supports researchers and industrialists for the development of innovative products.

- 3. Prevents the promotion of false marketing claims.

- 4. Ensuring quality of the product.

- 5. Ensure use of permitted additives.

There is a clear overlapping between labeling of conventional food and food supplements including energy bars/liquids. This overlap still becomes complex when we take into consideration on functional foods and nutraceuticals. Moreover, the current definition of nutraceuticals is also overlapping with the regulatory description for food supplements [6]. The initial step in determining the effectiveness of nutraceuticals is to separate them from the other food‐derived products, particularly food supplements, which does not fall under the regular diet of individuals [7]. These all lead to a criterion for setting up of a common regulatory framework for nutraceutical supplements, which can set a global standard to ensure the proper claims and quality of the product.

19.3. Overview on different regulatory bodies

There are different region/country-wise regulatory bodies looking for providing the insights into various parameters in region wise. The regulatory bodies are mainly concentrating on various aspects such as the use of specific raw materials, impurity limits, residual solvent, mycotoxins, and various food allergens.

19.3.1. Regulation in the United States of America

In the United States of America, the US Food and Drug Administration (US FDA) holds the regulation on the supplements, which are marketed as dietary supplements under the Federal Food, Drug, and Cosmetic Act as amended by the Dietary Supplement Health and Education Act of 1994 [8]. The FDA defines a dietary supplement as any product, other than tobacco that are intended to supplement the diet and that contain one or more of the following dietary ingredients: a vitamin; a mineral; a herb or other botanical; an amino acid; a dietary substance, for use by man to supplement the diet by increasing the total dietary intake; or a concentrate, metabolite, constituent, extract, or combination of any ingredient described previously FDA 2019. FDA also specifies the mode of administration for dietary supplements as for ingestion, and not by other means such as inhalable, injectables, etc., and also should not contain any ingredients which were previously classified as drugs. The US FDA allows the launch of dietary supplements by the regulation of postmarket surveillance of those ingredients which were in use before 1994. In the case of new dietary ingredients that were introduced after 1994, it has to go through a safety review through New Dietary Ingredient Notification process, even though there are no exceptions and as per the law, the supplements are not indented to diagnose, prevent, cure, mitigate, and treat diseases [9]. FDA regulates the use of botanical ingredients which are intended to diagnose, cure, or prevention of any clinical disorders in a different manner. In the case of such ingredients, it should follow appropriate premarket approval process [10]. All the supplements which are marketed in the United States should carry a label carrying the disclaimer that claims have not been reviewed by the US FDA. Office of Dietary Supplements (ODS) and National Institutes of Health also relates to support, conduct, and overview scientific support and intellectual leadership to strengthen the use of dietary supplements. The initiative by ODS on the use of omega-3 fatty acids [11], iodine [12], vitamin D [13], and iron [14] has laid an awareness among the public on the usage of the ingredients.

19.3.2. Regulation in Canada

In the Canadian region, most of the natural products are classified as a subclass of medicine, “Classified as Medicines” are defined as natural health products (NHPs) and is governed by Health Canada (HC). In Canada, NHPs are regulated under the governing body of Natural Health Products Regulations (NHPR), which came into force in 2004 [15]. The NHPR guidelines provide the need for product and site licensing requirements, including good manufacturing practices (GMPs), adverse reactions, labeling, product recalls, etc. [16]. HC also points on the utilization of healthiness claims for ensuring safety and efficacy by a clinical trial, scientific literature, and pharmacopeia [17]. Health products need to apply for a premarket authorization and submit the necessary information for evaluation by HC before launching the product in the Canada market, thereby allowing the authorities to have a close monitoring to authorities to have manufacturing and in addition to the postmarketing process. Manufacturers should also report on the dose, potency, ingredients, and the recommended use. HC also indicates the requirement of meeting the label requirements and adverse reactions [18]. There also exists a massive overlay among the herbs and the food. In Canada, herbal products can be vended with prevention and healing claims, with the exemption being which are marketed for serious disorders. HC also has started developing a framework for supplemented foods. The present Temporary Marketing Authorization framework for supplemented foods is aimed to gather data for the development of evidence-based regulations on food supplements on a future perspective [19].

19.3.3. Regulatory aspects in Australia and New Zealand

In Australia, many of the natural products such as herbals, vitamins, minerals, and nutritional supplements are treated as “complementary medicine” and termed as “Therapeutic Goods” [20]. These products are regulated under the Therapeutic Goods Act (TGA), which was enacted in 2004. The Australian authorities provide the Food—Medicine Interface guidance to characterize a product. The products that come under TGA are regulated by the TGA at the Federal level; however, the claims are regulated by state authority and region-wise food governing bodies. The TGA also has a list of “low-risk medicines” and “high-risk medicines.” As per TGA guidelines, low-risk medicine needs to be listed and high-risk medicines need to be registered. For registered medicines, they must be subjected to quality, safety, and efficacy evaluations [21]. Most of the products come under the category of listed medicines, while approximately 40% come under registered products. For products which are “classified as supplements,” therapeutic claims can be made but should label that these are not evaluated by the TGA.

In New Zealand also, there are two categories of products viz, Classified as Supplements or Medicines. In the regulatory point of view, dietary supplements are regulated by the Dietary Supplements Regulations of 1985. Dietary supplements are not required to submit for premarket approval and are supposed to be taken orally with the recommended dosage mentioned on the label and should not be intended for therapeutic purposes [22]. The class of dietary supplements also contains ingredients derived from animal/animal product origin, while these required to follow either the Animal Product Act 1999 or Food Act of 2014. The country also has a supplemented food standard that sets out the need for supplements that are marked as foods but have a modification in the additional ingredients added into it, so as to perform a physiological role as defined in the New Zealand Food Safety (2019) as “Supplemented Food” [23]. The country has also set up a national bill that will supersede the Dietary Supplements Regulations 1985 once approved [24].

19.3.4. Regulations in China

In the year 1995, the Chinese government introduced the Food Hygiene Law and established that the Ministry of Health is the statutory body for implementing the law and also declared that National Center for Health Inspection and Supervision as responsible for the approval of health foods licensing [25]. The law was divided into 104 articles. In the year 2003, a new regulatory body named State Food and Drug Administration was formed and consequently more than 4000 functional foods were approved, out of which some products are imported from other countries [26]. In 2009, the Food Hygiene Law was replaced with Food Safety Law [27]. In addition to the functional food, dietary supplements are also considered as nutraceuticals, which include vitamins, minerals, amino acids, fungi, probiotics, nucleic acids, and extracts of animal and plant origin [28]. In China, the food law was enacted in the year 2015 and established a new centralized system under the National Medical Products Administration [29]. Under the Chinese food safety law, the herbal products fall under the class of either “Classified as supplements” or “Classified as Medicines.” The category which is referred to as supplements has its roots in the Traditional Chinese Medicine [30].

19.3.5. Regulations in Japan

The recent 2015 regulation in Japan is mainly adopted from the United States with a slight exception. In Japan, the health claim systems came into practice as a result of developments in the scientific knowledge, global trends in the identification of health claims, and the occurrence of health hazards. The evolution of regulation of health claims in Japan started in the year 1952 with the establishment of special nutritive foods, which consisted of two categories: “foods that can supply certain nutrients” and “foods that are suitable for a particular purpose, which was followed by the proposal of the concept of functional foods by the Japanese academia in 1984. In the year 1991, Japan introduced the establishment of foods for specified health uses (FoSHU) [31]. FoSHU are foods which include functional ingredients and can affect the structure/function of the body. These foods are supposed to maintain or regulate specific health conditions such as gastrointestinal health, metabolic disorders, mineral absorption, bone health, and tooth health; however, claims of such products are approved on a regular basis [32]. For registration of new products, overseas applications are to be submitted directly in the portal of Ministry of Health, Labor, and Welfare (MHLW) Japan. The applications are invited in four different quarters, which should include the necessary details such as safety, efficacy, processing, formulation, analytical method, and chemical and physical analysis, as well as other specifics. There are four different categories of FOSHU which include: Regular FOSHU, Standardized FOSHU, Reduction of Disease Risk FOSHU, and Qualified FOSHU. In the Regular FOSHU, it includes consumer products, whose safety and efficacy regarding health claims have been proven by a series of safety/stability tests and clinical trials and have been approved by the MHLW. The Standardized FOSHU represents foods that contain certain active ingredients that are proven to meet the standards and specifications for a specific health claim, ingredient, and/or quality standards. If the efficacy of the product has been medically and nutritionally established, these supplements can be classified under Reduction of Disease Risk FOSHU. If the scientific data of the ingredients and products are not well known and are not clinically evaluated, the products can be classified under Qualified FOSHU [33]. Another category of functional food in Japan is the Food with Nutrient Function Claims (FNFC), which was enacted in April 2001. The FNFC includes all foods that have been labeled with nutrient function claims following the standards of the MHLW. The FNFC includes 21 ingredients including 13 vitamins, six minerals, and three fatty acids. The major difference between the FNFC and the FOSHU is that the former does not require any permission from the authorities as long as the manufacturer guarantees that the product meets the standards and specifications, unlike the accreditation and validation process under FOSHU. The FNFC rule does not require the need for having a specific logo on the label but should include the warning indications [34]. Japan also started a new regulation, which was enacted in the year 2015, focusing on the Food with Functional Claims. In this category, the manufacturers are allowed to label the food, if the business has submitted a notification to the governing bodies and indicating that the products meet national requirements, conform to the safety standards, and show scientific evidence for the functions claimed, obtained from clinical trials with humans to test their effectiveness or from the result of systematic reviews of human studies [35].

19.3.6. Regulation in the European Union

Of the global regulation applicable to the functional foods and nutraceutical supplements, the most stringent rules are applicable in the European Union (EU) region. European Food Safety Authority (EFSA) is an independent agency funded by the EU, which provides independent scientific support and communicates on existing and emerging risks associated with the food chain [36]. It came into existence in the year 2002 and was established in Parma, Italy. In the year 1997, Green Paper on Food Law preceding the food law of 1990 gave a new input to the foundation of European Food Law. EC No 178/2002 defines “food” or “food stuff” as “any substance or product, whether processed, partially or unprocessed, intended to be, reasonably expected to be ingested by human. The definition of food/food stuff includes beverages such as drinks and water, chewing gum, and any substances intentionally incorporated during different stages of processing. It excludes the category of feed, live animals, cosmetics (as defined by the Council Directive 76/768/EEC), tobacco and tobacco products, narcotic or psychotropic substances, residues and contaminants, etc.” [37]. Article 2A of Directive 2002/46/EC amended by the commission directive 2006/37/EC defines Food Supplements as “concentrated sources of nutrients or other substances with a nutritional or physiological effect, whose purpose is to supplement the normal diet, used alone or in combination” [38]. These are marketed in “dose” form, for example, as capsules, pills, pastilles, tablets, or liquids in measured doses, or released by drop-dispensing bottles, ampoules, or sachets of powder, among others and may be used to correct nutritional deficiencies. The list includes vitamins, minerals, amino acids, enzymes, pre- and probiotics, essential fatty acids, fibers, botanicals and botanical extracts, and miscellaneous bioactive substances [39]. Companies wishing to launch a nutrient source not included in the permitted list have to submit an application to the European Commission and as per EFSA directive 2002/46/EC, EFSA then prepares a scientific opinion to support the European Commission's evaluation of the request and based on the inputs from EFSA, the European commission reviews and updates the list of ingredients which can be used in food supplements [40]. If any substance, intended to be used in food supplements does not have the history of safe use in the EU before 1997, EFSA is requested to provide a scientific opinion on its safety according to Regulation (EC) No 2015/2283 on novel foods [41]. The European commission also has the right to list ingredients which are not allowed to use or its use is banned in foods. This procedure is listed in Article 8 of Regulation (EC) No 1925/2006, and the list of substances is listed in Annexure III [42]. EC 2002/46/EC also had laid down the mandatory labeling requirements, including the names of the categories of nutrients or substances that characterize the product or an indication of the nature of those nutrients or substances, the portion of the product, a warning note on not to exceed the permitted dosage level, etc. [43].

19.3.7. Regulation in the Association of South East Asian Nation region

All the 10 countries which constitute the Association of South East Asian Nation (ASEAN) countries have almost the same cultural and regional diversity in food, health status, and nutritional status. Regulatory agencies are stepping forward to set up several guidelines and process harmonization to have a uniform regulation throughout the nations [44]. All the regulatory requirements in ASEAN countries are handled by Traditional Medicines and Health Supplements Product working groups, ASEAN Economic Committee, and regional or state body of the country [45]. All the products which need to be marketed under ASEAN region need prior approval form marketing authorization by regulatory authorities in the respective member states [46]. There are some limitations also in setting a uniform regulatory platform throughout the nations. In Indonesia, these hurdles include the complex registration process, the presence of local manufacturers, and price sensitivity of the products [47], while when it comes to the Thailand market, the exporter needs to have a tie up with local distributor [48]. The common data requirement for registration of the products in these countries include [49]:

- a. Application form for registration.

- b. Copy of manufacturing license/import license.

- c. Power of attorney.

- d. GMP certificate for the manufacturing facility in the country of origin.

- e. Free sale certificate issued in the country of origin (Need to be the same as the certificate available in the country of origin).

- f. Product information, including drug interactions, use indications, dosage, overdose management, storage conditions, and shelf life.

- g. Regional language inserts including English language.

- h. A detailed description regarding the manufacturing process, as well as in-process quality control procedures.

- i. Stability data (in real time) from three batches.

- j. Quality specifications, including the relevant analytical methods for finished products, raw materials, and excipients.

- k. Three samples of the product, including Certificates of Analysis, active ingredients, and excipients.

19.3.8. Regulation in India

The Indian nutraceutical market is greatly dominated by pharmaceutical and food manufacturing companies with some exceptions, where companies focus on delivering value-added nutraceutical products. In 2006, the Government of India passed the Food Safety and Standard Act (FSSA) to integrate and make a guideline covering nutraceuticals, foods, and dietary supplements. The rule also signifies the importance of systems checks and balances, product recall procedures, and penalties to be implemented [50]. Under the FSSA, various food safety laws such as the Prevention of Food Adulteration Act (1954), Fruit Products Order (1955), Meat Food Products Order (1973), Vegetable Oil Products Control Order (1947), Edible oils Packaging Regulation Order (1988), etc. [51]. To make the FSSA more effective, an independent statutory body named Food Safety and Standard Authority of India (FSSAI) was enacted in the year 2008. After the implementation of FSSAI, regulations were set up for licensing and registration of food business, packing and labeling, food products standard and business, etc. [52,53]. This rule laid down a single legislation and specified authorities to regulate, manufacture, sell and distribute nutraceuticals, functional foods, and dietary supplements [54]. For registration of nutraceuticals in India, the business operator has to submit the below documents to the central licensing authority:

- a. Form B duly signed by the authorized signatory body.

- b. Blueprint/layouts plan of the processing unit.

- c. List of directors with the full address and contact detail.

- d. Name and list of equipment and machinery.

- e. Photo ID with address proof issued by government.

- f. List of food category desired to be licensed.

- g. Water analysis report.

- h. Proof of possession premises.

- i. Food Safety Management System Plan.

- j. No objection certificate.

- k. Food Recall Plan.

- l. Directorate General of Foreign Trade certificate of importer/exporter

- m. No objection certificate from the local body.

FSSAI regulations will cover eight categories of functional foods, namely, Health Supplements, Nutraceuticals, Food for Special Dietary Use, Food for Special Medical Purpose, Specialty food containing plant or botanicals, Foods containing Probiotics, Foods containing Prebiotics, and Novel Foods [55]. The FSSAI List also covers the list of ingredients and additives, which are permitted to be used in India.

19.4. Conclusion

Setting up guidelines for the nutraceuticals in the global scenario will allow the regulatory bodies to set up an international standard for launching the products and maintaining the qualities which adheres to the country regulations. These governing bodies can set up the guidelines on region-wise to meet the country regions. Going forward and taking into account on the market potential of nutraceuticals, there needs to set up a uniform guideline for launching the nutraceutical supplements on a global scenario, which will also ensure the manufactures to set up a uniform product on the global market.

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Chapter 20: Current trends and technologies in nutraceutical industry

J.A.A.S. Jayaweera
Department of Microbiology, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Colombo, Sri Lanka

Abstract

Because poor nutrition is a key risk factor for chronic diseases, many consumers desire to play an active part in disease prevention. Nutraceuticals, as opposed to pharmaceuticals, are food-based compounds utilized for illness prevention and therapy. Nutraceuticals are used to promote overall health and to prevent diseases. Nutraceuticals are getting more popular around the world as they form a part of people's everyday diet. A rise in noncommunicable diseases, high health-care cost, high personnel income, change of lifestyle, thinking pattern of the people and use of nutraceuticals as medicines, misbelief toward conventional medicines, and demand for the convenience are current trends in global nutraceutical industry. The development of functionally enriched foods necessitates the integration of several variables under consideration, and the final food products are influenced by the processing. The most recent techniques are now being used in both research institutes and the food industry, with the promise of reducing processing times, controlling Maillard reactions, improving product quality and increasing functionality. The use of technologies, in conjunction with other food industry trends and practices such as nanoencapsulation, food waste recovery, rising need for innovations, has resulted in new developments, data, and state-of-the-art in the sector. However, in depth studies are required to assess the benefits and harms of nutraceuticals.

Keywords

Current trends; Modern technologies; Nutraceutical; Safety and benefits

20.1. Introduction

The poor nutrition is a major contributor for many chronic diseases; many consumers desire to play an active part in disease prevention and the management. And that is where nutraceuticals come in between markets and the scientific domain of pharmaceutical companies. The term nutraceutical refers to natural sources that have health advantages in addition to the fundamental nutritional content found in food. Use of nutraceuticals would promote the overall health and are capable to prevent diseases. Nutraceuticals are divided into two categories: dietary supplements and functional foods [1]. Minerals, herbs, amino acids, enzymes, and a variety of other components can be found in dietary supplements in addition to vitamins. They are available in different forms, including gummies, beverages, energy bars, powder, and pills. Magnesium pills, multivitamin gummies, and protein powder are a few examples [2]. Functional foods include whole foods as well as fortified, enriched, or augmented dietary components. Traditional functional foods are nutrient-dense foods such as legumes and whole grains. Modified functional foods include juice with added calcium, cereal with added iron, and milk with added zinc and folic acid [3]. Nutraceuticals, as opposed to pharmaceuticals, are food-based compounds utilized for illness prevention and therapy. Nutraceuticals are often classed as a subset of food and, as such, are not as tightly controlled by the country's local regulatory body as pharmaceuticals [4].

20.2. Current trends to drive nutraceutical industry: global scenario

Nutraceuticals are getting more popular around the world as they form a part of people's everyday diet. The nutraceuticals global market is estimated to be worth around US$233.9 billion in 2020, with a projected value of US$358.5 billion by 2027. Japan and China are the second and third major users of nutraceuticals after North America. They account for around 78% of the total market in the Asia-Pacific area [1–3,5].

Nutraceuticals are becoming increasingly popular because of their potential nutritional and safety characteristics, as well as their therapeutic power. The food and pharmaceutical companies know well about the shifting patterns brought on by the benefits of these chemicals. Moreover, the majority of nutraceuticals have several therapeutic advantages [6].

20.2.1. Rise of noncommunicable diseases

When considering the trends toward (Table 20.1) the popularity and improvement of the nutraceutical industry, the rise of noncommunicable diseases in the world population would have significantly influenced. Globalization and urbanization have increased in developing countries, leading to an increase in noncommunicable diseases such as cardiovascular disease (CVD), cancer, diabetes, and lung disorders. Obesity, allergies, and Alzheimer's disease are also becoming increasingly common [5–7].

CVD is a group of heart and blood vessel illnesses that includes coronary heart disease, peripheral vascular disease, cerebrovascular disease (stroke), hypertension, and heart failure. A significant proportion of CVDs are avoidable, and numerous studies have emphasized the protective role of a vegetable and fruit-rich diet against CVD [8]. The regular exercise in conjunction with nutraceuticals such as vitamins, minerals, antioxidants, dietary fiber, and omega-3 polyunsaturated fatty acids is indicated for CVD prevention and therapy. Polyphenols would prevent coronary artery disease by altering the metabolism and communication in the cells [6–8]. Flavonoids in the forms of flavanones and flavanols are abundant in vegetables, onions, endives, grapefruits, apples, and red wine and have an important role in CVD prevention and treatment. Flavonoids inhibit angiotensin-converting enzymes and cyclooxygenase enzymes, which involve in the metabolism of prostaglandins and aggregation of platelets. Also, they safeguard the body circulation, which transports oxygen and essential elements to cells [9]. Zingiber officinalis rhizome is a common condiment for a variety of dishes and beverages. It has a long history of therapeutic use and has been shown to reduce the risk of CVD. Phytosterols prevent the absorption of dietary cholesterol and increase the elimination from the body. As a result, they have the potential to lower CVD morbidity and death. Phytosterols are found in most plant seeds, and green and yellow vegetables contain substantial amounts of sterols. The omega-3 series found in fish influences the plasma lipid concentration and prevention of CVD [4,5,10].

Cancer is a significant public health issue in most countries. The world cancer report reflects that the rates of cancer are rising, with 15 million additional cases expected by 2020, representing a 50% increase. Cancer prevention can be aided by a healthy lifestyle and nutrition. The phytochemicals such as carotenoids are responsible for the color of foods. They pose antioxidant properties and can help to prevent neoplasms. The relevance of lycopene in cancer disease has sparked renewed interest in carotenoids [4,6,10]. Lycopene-rich vegetables and fruits protect against cancer by lowering oxidative stress in turn minimizing the damage of cellular DNA. Citrus fruit flavonoids can help prevent cancer by acting as antioxidants. The compounds in tea epigallocatechin gallate, curcumin from curry, and isoflavones in soya are unique sources in cancer prevention [11]. Saponins pose antimutagenic and antitumor properties, and they may reduce the risk of human cancers by preventing cancer cells from proliferating. Saponins are present in peas, soybeans, and some herbs with names that allude to their foaming qualities, such as soapberry, soapwort, and soapbark. Tomatoes, potatoes, alfalfa, spinach, and clover also contain them. The great amount of saponins for food industry are derived from Yucca schidigera and Quillaja saponaria [9,11]. Sulfur obtained from garlic would stimulate the immune system, lower atherogenesis, platelet aggregation, and cancer risk. Broccoli contains a lot of sulforaphane, which induces the phase-2 enzymes. It creates D-glucarolactone, a powerful antibreast cancer agent, and the antioxidant sulforaphane would stimulate the activity of endogenous detoxification enzymes [12]. Large-scale clinical trials indicate that to prevent prostate cancer, medicines such as green tea, vitamins D and E, selenium, lycopene, soy, and inhibitors of 5a-reductase are useful. However, in smokers, carotene, N-acetylcysteine, tocopherol, retinol, retinyl palmitate, or isotretinoin would not prevent cancer [13].

Table 20.1

Current trends to drive global nutraceutical industry.

Rise in noncommunicable diseases
High health-care cost
High personnel income, change of lifestyle, thinking pattern of the people, and use of nutraceuticals as medicines
Misbelief toward conventional medicines
Demand for the convenience

In recent years, multiple nutraceuticals have been scientifically established in preclinical research to improve type-2 diabetes mellitus. Isoflavones are phytoestrogens that are structurally and functionally comparable to human estrogen [11–13]. The antioxidant lipoic acid is useful for patients with diabetic neuropathy and in long run it would be an effective dietary supplement to prevent further complications. To reduce body weight, attain glycemic control in patients with diabetes and reduce blood cholesterol, the dietary fibers obtained from psyllium are used. The extracts of Teucrium polium, cinnamon, and bitter melon have been demonstrated to prevent and treat diabetes [14].

The significant increase of viral infections, as well as the accompanying death rate, has created a need for effective and safe antiviral medications. Over the years, the antiviral pipeline has been dried following the emergence of multiple drug-resistant pathways, availability of a limited number of viral targets, the rapid viral gene evolution, and the mutations leading to the emergence of new viral strains [11–13]. Nontraditional medicines, nutraceuticals, can assist in combating the threat. These agents are not only having virucidal capabilities but also they can promote natural immunity and health by replenishing glutathione levels and reducing the amount of free radicals in the cells [14]. As a result, the viruses find it more difficult to divide within the host body, and the severity of the clinical presentation reduces, leading to a low fatality rate and a cure. In addition to eliciting an effective immune response, probiotics would bind to viruses in the gut, restricting those from adhering to the host cell via a variety of mechanisms, including steric hindrance, receptor-specific competitive and noncompetitive binding, and the bonding of certain other compounds to avoid binding [15]. Nutraceuticals with antiviral and immunomodulatory effects include resveratrol, quercetin, curcumin, epigallocatechin gallate, N-acetyl cysteine, and palmitoylethanolamide. The natural agents provide a protection against coronavirus by producing antivirals such as nitric oxide, free radicals, and stimulating CD8+ T lymphocytes, CD4+ T lymphocytes, phagocytosis, and B lymphocyte proliferation [2,3,11–14].

Diets from plant-based products are demonstrated to reduce the risk of contracting influenza and pneumonia as they are rich in dietary fiber, antioxidants, and vitamins, which aid to lower the body weight and enhance the immunity. However, the benefits associated with nutraceuticals must be researched in order to generate persuasive scientific evidence of their safety and efficacy [1,3,15]. Lactobacillus rhamnosus is safe for premature newborns to consume, demonstrating that probiotics are safer alternatives for interventions-related therapy in all age ranges. Immunocompromised people as a result of chemotherapy or a critical disease, on the other hand, should use probiotics with caution. Probiotics have been proven in studies to lower the severity and duration of acute gastroenteritis following Rota virus and acute viral respiratory tract infections [16].

Obesity is a risk factor for a variety of diseases, including hypertension, congestive cardiac failure, unstable angina, dyslipidemia, respiratory diseases, osteoarthritis, malignancies, thrombosis in renal vein, and subfertility. Obesity is a worldwide problem; thus, proper diet and regular physical activities are crucial in its prevention and control. Nutraceutical therapies are currently being researched extensively as potential treatments for obesity. Capsaicin, conjugated linoleic acid, Momordica charantia, and psyllium fiber are examples of antiobesity nutraceuticals [14]. An effective nutraceutical that can increase metabolism or decrease the intake of calorie is desired for weight loss. Herbal stimulants that aid in weight loss include caffeine, ephedrine, chitosan, ma huang-guarana, and green tea. However, due to the possibility of adverse outcomes, its use is debatable. The extract from green tea and 5-hydroxytryptophan would be useful for weight loss; the former boosts metabolism, while the latter suppresses hunger [3,4,15].

Although there is not yet enough scientific evidence to prescribe nutraceuticals for Parkinson's disease, exploratory trials have indicated some promising outcomes. Regular use of vitamin E, glutathione, and creatine appears to be preventive of Parkinson's disease [14–16]. Among elders, the Alzheimer's disease is the commonest form of dementia and it is characterized by a gradual loss of cognitive abilities. Chemical formulations usually work by blocking the activities of the enzymes that cause Alzheimer's disease. However, with time, these pharmacotherapies develop a variety of side effects, including toxicity in several organs. Curcumin, resveratrol, and genistein are multipotent natural substances that can inhibit the development and deposition of amyloid plaque peptides. These natural chemicals have also been discovered to modulate several intracellular signaling molecules and enzymes such as β-secretase and γ-secretase [17,18].

The linoleic acid and omega-6 essential fatty acid are the elements of gamma linolenic acid (GLA). GLA is a nutraceutical that is useful to alleviate inflammation and autoimmunity. GLA is converted to dihomo-gamma-linolenic acid, which is then oxidized by lipoxygenase and cyclooxygenase enzymes to create antiinflammatory eicosanoids. Trace amounts of preformed GLA can be found in nuts, green leafy vegetables, and vegetable oils such as seed oil, borage oil, Oenothera biennis oil, black currant oil, and hemp seed oil [17,18]. There are also herbal nutraceuticals with antiinflammatory properties. Gentianine, found in Gentian root, is a powerful antiinflammatory substance. Bromelain, enzyme with proteolytic properties is found in stinging nettle, turmeric, pineapple, teas, and curcumin preparations, poses an antiinflammatory action [16,17].

Probiotics are one alternative medicine being researched for the treatment of allergies, particularly food allergies. Probiotics can be found in foods such as yogurt, milk, and supplements. They are supposed to boost the immune system by introducing helpful bacteria into the digestive tract. While it is unclear whether probiotics can aid with allergy treatment, there is probably no harm in taking them [17,18].

In healthy individuals, angiogenesis, an enzymatic process is normally suppressed. Antiangiogenic medications may not generate detrimental effects even after prolonged durations of exposure, since they selectively act on newly formed blood vessels while sparing older ones. Antiangiogenic substances may help to avoid degenerative diseases such as multiple sclerosis, arthritis, and osteoporosis. Also, they are effective against diabetes mellitus, malignancies, Alzheimer's, and Parkinson's. Curcumin, flavins, isoflavones, catechins, resveratrol, proanthocyanidins, flavonoids, saponins, terpenes, chitin, chitosan, vitamins B3 and D3, fatty acids, peptides, and amino acids are examples of bioactive chemicals that may be effective angiogenic agents [1–4,16,17].

The drug toxicology is complex; there is strong evidence that oxidative stress plays a role related to adverse effects of a wide range of medications. The majority of plants and its extracts have antioxidant activity, and in addition to numerous methods of combating poisons and synthetic medications, they can generally lessen their toxic potential by minimizing the oxidative stress. The renal and hepatic activities are more crucial than other organs in the process of alleviation of harmful effects of medications and poisons. In this regard, numerous researches have been conducted to investigate the protective properties of nutraceuticals, particularly herbal plants, against harmful compounds and other medications, with encouraging results [6,9,14].

20.2.2. High health-care cost

Health spending is growing faster than expected of the world economy, amounting for 10% percent global gross domestic product. A new report of the World Health Organization on global health expenditure finds a rapid upward trajectory of global health expenditures, notably in low- and middle-income nations, where health spending is expanding at a rate of 6% per year on average compared to 4% in high-income countries [4,9,18].

Out-of-pocket expenses, as well as resources such as insurance schemes, employer-provided health-care programs, and nongovernmental agency operations, all contribute to government spending on health. Governments fund an average of 51% of a nation's health-care expenditures, while out-of-pocket costs account for more than 35% of overall health-care spending. As a result, one billion people are driven into abject poverty each and every year [16–18].

Government health spending per capita in middle-income countries has more than doubled since 2000. In lower-middle income countries, governments spend a total of US$60 per person on health, while in upper-middle income countries, they spend about US$270 per person. People are less likely to fall into poverty while pursuing medical services when the government invests more on wellness. Government spending, on the other hand, can only decrease access gaps if expenditures are correctly tailored to guarantee that the entire population has access to care [14–16].

Consumers seeking to enhance their own health without incurring the costs of medical treatments are turning to the nutraceutical business for assistance. Fortified foods and beverages are fashionable in health-conscious individuals looking at other options to over-the-counter and prescription medications [18–20].

20.2.3. High personnel income, change of lifestyle, thinking pattern of the people, and use of nutraceuticals as medicines

Because of the rise in the number of individuals with high incomes, the emphasis has turned to holistic health, and in the search for immunity, the prescription of such pharmaceuticals has increased several folds [14–16]. Changing lifestyles and sedentary work environments, along with poor food habits, have necessitated the use of dietary supplements to replace micronutrients and maintain optimum health. Some customers feel that an unhealthy diet will result in an increase in spending on regular pharmaceuticals, which could be one of the factors driving up demand for nutraceuticals. In fiscal year 2019, India's net disposable income was approximately 1.7 trillion Indian rupees. When compared to prior generations, the younger generation may spend more on health products [21]. The growing concern about nutrition, as well as increased understanding and easy access to information, has resulted in a growth in the usage of health supplements and nutraceuticals. The growing awareness of the importance of immunity and dietary supplements has resulted in a huge increase in demand. Even after the COVID-19 pandemic has subsided, behaviors such as using immune-boosting nutraceutical supplements may persist [21,22]. The key consumer-related behaviors that are transforming the nutraceutical market include changing lifestyle and increased awareness. The epidemic has given the nutraceutical business a boost, with a focus on prevention and general well-being [9].

20.2.4. Misbelief toward conventional medicines

Pharmaceutical medications and their negative effects are causing increasing concern. With nutraceuticals viewed as a more natural option, the use of nutraceuticals is likely to rise in the coming years [9–11].

20.2.5. Demand for the convenience

Customers prefer ease. Boomer generation does not want to keep swallowing pills, and youngsters prefer products that can be utilized when out and about. Nutraceuticals can also be found in the form of gummies and effervescent tablets for those who dislike taking pills. Protein bars, dry nuts, grains, and bars are examples of nutraceutical snacks that are ideal for on-the-go munching [12]. The nutraceutical industry has shown potential for growth, particularly in the context of elders and continuous incline of cost of health care. Nutraceutical producers, retailers, and exporters must grasp the drivers of nutraceutical market growth in order to thrive [22,23].

20.3. Technologies in nutraceutical industry

The development of functionally enriched foods necessitates the integration of several variables under consideration. Separation methods, toxicity evaluations, and tests of stabilization and function are examples of these. The final food products are influenced by the processing. The technologies used can have an impact on the composition and efficacy of nutrients; for example, as nutraceuticals are processed, stored, and transported, the elimination of active chemical molecules or the decrease of their activity generally increased [24].

Nonthermal advancements such as ultrasound, high hydrostatic pressure, pulsed electric field, high voltage electrical discharge, and cold plasma have the potential to treat foods without demolishing the important nutrients and multisensory attributes that are generally disrupted during thermal processing. New techniques are increasingly being applied in both research facilities and the food processing industry, with the promise of reducing production time, managing Maillard events, improving the quality of products, and improving usefulness [22–24].

The technology in the field, in combination with other food industry systems and development, such as nanoencapsulation, food waste recovery, and the increasing need for innovations, has resulted in new breakthroughs, information, and state-of-the-art in the sector. Despite substantial improvements in the detection, separation, and descriptions of these constituents, there is still a demand for prime and near-market platform technologies for converting those constituents into marketable real worth food products and nutraceutical formulations [24,25].

A range of factors influence the effectiveness of dietary bioactive, including metabolomics, nutrigenomics, bioavailability, and stability within food material. Excess pH, the mucous membrane, the epithelium, and enzymes in the digestive tract, for example, impede food component distribution via the oral route. Functional foods are made by combining old or innovative food products with compounds that change their properties such as by changing the structure, binding, or interface and provide health benefits [3,4,24–26].

Food preparation necessitates the assessment of various parameters such as the examination and interpretation of sources, the recognition of biomolecules, the selection of recovery approaches and technologies, the performance of toxicological tests when necessary, and finally the assessment of bioaccessibility, bioactivity, and stability. Without the requirement for bioactivity tests, bioaccessibility assays evaluate nutritive benefits of foods, when contrasted to health claims of foods. Two of the most essential factors determining dietary component bioavailability are interactions between active molecules and the source of origin. Conjugated or free carbohydrates are typically found in materials derived from plants. Proteins, dietary fiber, and fats, on the other hand, are typically associated with antioxidant in bunches inside the fruit tissue [26,27].

Bioaccessibility and bioactivity are not usually well understood when studying the effects of bioactive on human health. The most abundant bioactive in vegetables, fruits, and other foods, in particular, do not have to result in the most active constituents in tissues. Furthermore, food processors frequently assert that some of their products have a good attribute, but these assertions must be validated by legitimate study. Many scientific papers, for example, remark that the majority of the beneficial effects of food components' bioactivity have been shown in laboratory tests rather than in vivo studies, which are often difficult to conduct [27–29].

The food business concentrates primarily on keeping the product's sensory properties such as taste, flavor, smell, and color during bioactive chemical fortification. However, it is equally critical to keep these molecules effective. The first step is to determine the appropriate amount of bioactive component to be included to the food in order to prevent some illnesses following eating [16].

The next step is to look into the systems needed to safeguard bioactive molecules not just in the food matrix but also during consumption. Bioactive delivery systems in the food industry are designed similarly to pharmaceutical delivery systems, with the goal of increasing solubility, so that they can hit their target absorption area via the circulatory system, trying to control the dissolution rate, and masking any unpleasant organoleptic properties [30–32].

It is also crucial to address bioactive material stability during manufacturing, treatment, encapsulating, and shelf life against physiological and biochemical degradation. Solubility and bioavailability of food components can be improved by reducing particle size and using appropriate compositions that protect bioactive from enzyme-catalyzed breakdown until they reach the intended site in the gastrointestinal system. Emulsions made from peptides, biodegradable polymers, fats, or hydrogels are frequently used in these formulations. Low molecular weight surfactants in micro, nano, and micellar frameworks, egg or sunflower made liposomes, PEGylated liposomes, organogels, or tea saponin have all been presented as appropriate carriers [17,19,32].

Processing technologies can have an impact on the functional qualities such as biological activity and its accessibility of nutraceuticals and, as a result, their therapeutic effects. The loss of food bioactive is proportional to the extent of food processing and the length of storage and transit. Controlling the degree of disintegration of fruit and plant tissues is so critical, since it influences component functionality as well as the degradation and quality of foods. Since the demand for nutraceuticals has grown, researchers and the nutraceuticals platform have increased their efforts to improve the processes of processing that preserve goods' qualitative, active, and nutritional properties [21–23,32].

Pressing mechanically, milling in wet, and filtration using membranes are three common food processing processes for removing water, softening tissue, and removing particles and fats, respectively. Ultrafiltration, isoelectric, and alcohol precipitation are all standard techniques for separating macromolecular components like pectin and polypeptides from wines and various juices. Concurrently, acid hydrolysis is often used as a preparatory method, while solvent extraction is used to recover antioxidants and glucosinolates [24–26,32]. The technologies, in hand with heat-derived production such as spray drying and pasteurization, are commonly used to manufacture the fortified foods and novel nutraceuticals. All of these traditional approaches are well utilized in the food industry, despite several issues that limit their uses and present practical issues in many circumstances. Solvents of organic are typically neither green nor food-grade, boosting consumer understanding about the adoption of a safe food chain [17,19,23–25,32].

Food sterilization by heating and conventional cooking methods such as parboiling, steaming, and searing has long been employed in the food industry to soften plant tissues and then for food safe operation by inactivating microbes, fungi that ferment and limit food half-life. Similarly, many of the deleterious changes that results while storing food are accelerated by additional enzymes, especially pectin methylesterase, polyphenol oxidase, and peroxidize, all of which are inactivated by heat. Nonetheless, heat processing of food causes food constituents to be destroyed. Because of these oxidizing and isomerization processes, food quality degrades less and has a shorter shelf life. Furthermore, overheating the food matrix frequently increases undesirable Maillard processes, which reduce the sensory qualities of foods [15,17].

Food bioactives' bioaccessibility, functioning, and stability are also impacted. Other typical nonthermal processes that require high pressures such as nanofiltration and freeze-drying have higher energy demands and operational costs. Furthermore, traditional procedures for encapsulating dietary bioactives may result in dangerous products. Because of these constraints and disadvantages, food processors are looking for novel processing technologies that will allow them to preserve the functionality of bioactive chemicals as well as the qualitative qualities of foods. Consumers desire more custom-made foods with a longer half-life and a sense of “freshness,”as well as less kitchen preparation time and labor [12–14,23,33].

The most common developed techniques in the nutraceutical sector include high hydrostatic pressure, ultrasound-assisted and microwave-assisted extraction, pulsed electric field, radio frequency drying, high voltage electrical discharge, and supercritical fluids. Cold plasma treatment, electroosmotic dewatering, and nanoencapsulation are some of the other emerging technologies being researched by food manufacturers (Table 20.2). New technologies are not based on high temperatures, but rather on the production of heat via intrinsic energy transfer such as resistive and heat release with pulsed electric fields and high hydrostatic pressure, respectively, which minimizes the loss of sensory, nutritious, and health beneficial characteristics [33–35].

For example, processing by means of high pressure (200MPa) is widely used to destroy microorganisms by breaking their membranes or modifying the solubility of whey proteins. It improves plant permeation, allowing for more bioactive mass transfer and diffusion during phase transition. This approach enables to solubilize food materials in a rapid manner while preserving appearance, flavor, and consistency [23,26,32,34].

Supercritical carbon dioxide extraction is another process that involves the utilization of gas above its critical pressure. Despite being a fast and selective process, it has been largely advised to recover value-added compounds due to its high production costs (perfumes from natural sources and specific flavonoids extraction). Ultrasound is another nonthermal approach that has been demonstrated to expedite mass transfer by producing bubbles in food structures when employed in high-intensity (>10W/cm^2, 100kHz) mode [17,19,33–35].

Table 20.2

Novel and emerging technologies for nutraceutical production.

Novel technologies (nonthermal)
Extraction methods
High hydrostatic pressure
Ultrasound-assisted extraction
Microwave-assisted extraction
Drying methods
Pulsed electric field
Radio frequency
Emerging technologies
Cold plasma treatment
Electroosmotic dewatering
Nanoencapsulation

Low-intensity (1W/cm^2, 100kHz) modes are used throughout the food manufacturing to observe the procedures and acquire data on various properties, such as milk coagulation, the formation of bubbles in carbonated foods, pitting in cheese, quality of eggs, proportion of lipids in meats, snack creaminess, beverage fermentation control, fruit, and dough morphology. Ultrasound has been demonstrated to be particularly efficient against pathogens contaminating fruit juices, resulting in a 5-log decrease (FDA criteria) of Escherichia coli in fruit juices. In vitro bioavailability of lycopene is decreased during ultrasonic processing of tomato pulp due to the release of pectin during the incomplete deesterification [17,19,33,35].

In novel encapsulation, 10–100nm multiphase colloidal nanoemulsions are prepared using ultrasounds with the goal of delivering nutrients and bioactives in the dimension of nano, thus in turn increasing the bioavailability, stability, and energy, as well as controlled release to desired organ of the body [19,23].

Radiofrequency drying has been presented as an alternative choice among contemporary drying processes. With radiofrequency drying, evaporation of water occurs around 80°C with a faster production time, and the foodstuff matrix is treated uniformly. These techniques, however, are inefficient in terms of energy use. When compared to standard hot air drying, electroosmotic dewatering saves energy consumption by two-thirds [17,19,33]. It operates by exerting mechanical pressure to the interface of colloidal particles and aqueous suspensions while concurrently creating double layers. Expedited mass transfer has also been proposed using pulsed electric fields, which may destroy the cellular membrane, soften plant tissue texture, and electroporation but also increase the recovery of key compounds from diverse vegetables and fruits. Pulsed electric fields have been discovered to preserve nutrients in milk, eggs, and liquids and improve the packaging of bioactive compounds by increasing fluid bed clumping of soy protein isolate [17,19,33].

Another method that has been tested to extract compounds, which can use value addition from the seeds of grapes is high-voltage electric discharge. The extraction is put in a container between two electrodes that deliver 40–60kV/cm, 2–5s short pulses to disrupt food components. The food is similarly put between two electrodes in ohmic heating before an oscillating electric current is sent across the circuit [17,19,33]. Electrical resistance in food generates exceptionally quick and homogeneous thermogenesis. This method is comparable to heating using microwave, but without the intermediate step of transforming electricity into microwaves [34]. Ohmic heating is used for pasteurization, sterilization, and defrost the turbulence sensitive fluids and foods such chicken, fruits, fish, vegetables, and instant meals. The cold plasma is a quasineutral particle system made up of a mixture of highly active semigas and semifluid. Electric discharges utilized at various pressure levels in high-moisture foods generate cold plasma, while a vacuum helps the conversion of liquid into gas. This unique approach for inactivation of pathogens in food has been tested [17,19,33–35].

The technologies which are used during food processing effect the efficacy and quantity of active compounds. For example, when foods are prepared, preserved, and delivered, the functional characteristics of the active compounds would diminish. The use of growing technologies in the food business, the immediate need for sustainability practices, food scraps reclamation, have resulted in new information and biodistribution goals [4,7].

Fermentation of foods using bacteria is a common technique around, and it is regarded as an essential food preservation strategy. Fermented foods are offering pleasing flavors, smells, and textures and capable to improve human nutrition greatly [7,11]. Fermented foods and beverages are thought to account for roughly one-third of the human diet [3,6,13]. In addition, removal of glycol-side residues and toxins, inhibition of microbial pathogens in the foods, and the delivery of probiotics are outcomes of fermentation [9,14,21]. Traditional fermentation processes vary due to cultural and socioeconomic factors and technically would be more susceptible to contaminate with food pathogens. The concerns on the safety of foods are raised due to inappropriate conditions of fermentation or processes. As a result, there is a growing interest on standardization of fermentation techniques and microbes to increase the safety of food and benefits [12,32].

The awareness on the medical benefits of milk products following fermentation is an emerging area of nutrition and biomedical science research. The tolerance of lactose, reduced length of episodes of diarrhea, immune enhancement, serum cholesterol reduction, protection from malignancies, and mineral absorption of minerals are possible benefits of fermented milk products [34,35]. The effects of several strains of bacteria on Camellia sinensis (tea) fermentation provide a protection against Salmonella and Staphylococcus microbes. Tea fermentation would alter the quantity of organic acids and polyphenols in tea. The heath value addition of black tea following fermentation of yeast would reveal an accumulation of vitamin, caffeine decrease, excess tannins, and improved bronchodilator therapeutic efficacy. Toba et al. developed two patents describing an antioxidation of food product made from strains of Lactobacillus plantarum and natural material, which contains magnesium. A dry extract was obtained that would be mixed with dairy and consumed, and this would promote its natural nutritional and therapeutic benefits of fermented tea products [14,26,32]. Izvekova et al. developed Lactobacillus spp. cultures from the Lactobacillus acidophilus species, allowing for different variants of liquid milk infant formulas, solid for yogurt or ice cream, and powder additives for pharmaceuticals [22,28].

Consumption of beneficial microbes in the form of fermented foods has been around for millennia. Metchnikoff's longevity theory, developed in the early 1990s, paved the way for the present notion of probiotics and its commercialization. Over the years, our understanding of probiotics, prebiotics, and their combined synergistic activity, known as symbiotic, has expanded dramatically, and many potentials and even verified mechanisms of action for human health have been recorded [14]. Recent findings in human microbiota metagenomic research have convincingly revealed their potential to regulate the pathophysiology of numerous diseases. The inclusion of probiotics and prebiotics in the human diet benefits the intestinal microbiota, the unseen controller of human health. Interestingly, the use of probiotics and prebiotics in the development of novel functional foods is quickly expanding [32].

Recent developments in gut microbiome research have also led to the discovery of the therapeutic potential of additional probiotic candidates beyond the regularly used genera, primarily from adult microbiota dominating individuals. These microorganisms are known as next-generation probiotics, and several unique strains are thought to have next-generation probiotic properties. Their safety has yet to be properly proven, although they are more likely to be suitable for a pharmaceutical delivery route than delivering with a carrier food. Because probiotic distribution was originally related with food substrates, excluding carrier food matrices may have an effect on the potential synergistic effect. Post-biotics are bacterial metabolites or metabolic byproducts of gut microbes that try to imitate the favorable therapeutic effects of probiotics. The use of these bacterial metabolites in health promotion is a fascinating, but mostly unexplored, area. As a result, the goal of this special issue is to provide a venue for showcasing recent probiotic and prebiotic food innovations [17,19,33].

Bread, as a fermented food, depending on the types of cereals included and the manufacturing processes would provide various health benefits. For example, research has suggested that certain breads following in vitro fermentation may have chemopreventive (colon cancer prevention) effects. Several studies have been conducted to investigate the health benefits of nutraceutical-infused bread items and its physical and chemical aspects. There have been studies on breads with the addition of phenolics such as caffeic, gallic acids, and black currant. The effect of bread with prebiotics and antioxidants with tomato paste as a lycopene source on immunological parameters was investigated. The findings revealed a considerable increase in antioxidant properties. Bread supplementation with nutraceuticals such as beta-carotene has been investigated. It has also been reported that beta-carotene can be added to muffins [14–16,21,32].

The plant products such as roots, leaves, branches, blossoms, and seeds are employed in numerous formulations that are said to beneficial for human health by regulating the neurological system, heart, and circulation, as well as healing the immune, digestive, and respiratory systems [17]. Soda cracker biscuits have been made using a 0.5% basil fine powder and 0.01, 0.02, or 0.03% of BHA by weight. The findings demonstrated fine powders or extracts of plant could be utilized as a replacement for conventional antioxidants in soda cracker biscuits without compromising taste and the palatability [35,36]. A commercial bakery created a fenugreek bread recipe by mixing flour of fenugreek into a formula of wheat bread. The findings demonstrated that acceptable baked goods may be created with additional fenugreek, which could reduce resistance of insulin and best for type 2-diabetes [11,14,17].

Various cereal family species such as wheat, rice, corn, barley, and oats occur in a variety of geoclimatic zones and have evolved to a wide range of obstacles. Cereals serve an important role in feeding the world population because the endosperm is primarily made up of carbohydrates, making them the major source of energy. Most cereal germs are heavy in oil and lipid-soluble vitamins, whereas the outer layers are high in cellulose, hemicellulose, and minerals. Cereals include dietary fibers, phenolic compounds, carotenoids, anthocyanins, vitamin B and E, and micro- and macroelements of nutrition. Because of these health-beneficial elements, in the food industry, byproducts such as bran are recognized for their nutritional value [17,19,33].

The nonthermal technologies are efficient, rapid, and credible way to preserve the bioavailability of nutraceuticals, improve the protein gelling capacity, and increase the recovery of compounds with medicinal values. However, if used for an extended period of time, with a high intensity may cause proteins and carbohydrates breakdown and oxidation of labile lipids and glucosinolates. Therefore, monitoring and optimizing the parameters are vital.

Several of these mechanisms now have been proved industrially; however, more work in this regard is required. Novel techniques and nutraceutical processing trends have resulted in discoveries in the inclusion of components, fortification of foods, and, eventually, food consumption [9,11,17,21]. By means of value-added products, the nutraceuticals reach to consumers as sachet or ready to use formulations. Also, various capsules have been developed using technologies similar to the pharmaceutical industry [19,37]. Many individuals feel that nutraceuticals, particularly medicinal plants, are significant solutions for addressing health conditions that have no adverse effects. This idea stems as they used those for a long time without major toxicity. Perhaps, it is applicable for a wide range of nutraceuticals and normally they have fewer adverse effects than drugs. Those who practiced and consumed the conventional medicine believe that drug to be effective; it must unavoidably have adverse effects [14–16,22].

Herbal medications are considered pharmaceuticals by the medical system and they must pose adverse effects. As a result, those must be processed using the best components and used with prudence. Surprisingly mankind consumes a vast number of plant products by means of various products of nutraceuticals to meet their day-to-day nutritional requirements; however, only a limited amount of products have undergone extensive risk assessment [21]. Many are still little investigated and underdeveloped, and their wild counterparts are on the verge of extinction and in desperate requirement for conservation. The valuable plant resource stewardship would necessitate in-depth scientific studies [17,30–32]. The list of nutraceuticals under investigation is constantly evolving to reflect market trends, scientific evaluation, and interest of consumers. With the growing interest in nutraceutical intake, extensive study is required to ensure that the use of nutraceuticals is harmless and beneficial. Nutraceuticals could be beneficial to health, retards the process of aging, involve in chronic diseases prevention, and regulate structure and function of human body [19,32,37]. The mechanisms of the activities of nutraceuticals are not completely understood. They may, however, be engaged in a wide range of biological processes, such as signal transduction pathway activation, antioxidants-mediated protection, genetic expression, cellular proliferation, differentiation, and mitochondrial integrity [4–6,17,19,32–37].

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Chapter 21: Future prospective of nutraceutical and functional food with herbs and spices

Parveen Bansal
[1]
, Mukesh Maithani
[2]
, Vikas Gupta
[1]
, Gunpreet Kaur
[1]
, and Renu Bansal
[3]

1
University Centre of Excellence in Research, BFUHS, Faridkot, Punjab, India
2
MRU, VCSG Government Institute of Medical Sciences & Research, Srinagar, Uttarakhand, India
3
Government Medical College, Patiala, Punjab, India

Abstract

The health and well-being of human beings are highly dependent on the consumption of nutraceuticals and functional foods. They have acquired considerable importance for their alleged safety and possible nutritional and medicinal effects. Several studies have linked nutraceuticals and functional foods as useful in fighting a range of degenerative diseases. Therefore, a great deal of research has been observed in recent years on functional attributes directly related to the health benefits of various medicinal plants, plant extracts, phytochemicals, herbs, and spices. Although a large number of naturally derived health substances are plant-based, there are also a number of physiologically active components in herbs and spices that deserve attention for their potential role in optimal health. The consumption of biologically active ingredients in nutraceutical/functional foods has been linked to herbs and spices to help fight many diseases such as cancer, cardiovascular disease, obesity, osteoporosis, allergies, gastrointestinal disorders, and dental problems. Much research is still needed to validate the possible health benefits of herbal nutraceuticals/functional foods for which the relationships between diet and health are not sufficiently validated and to create a solid scientific information base for suitable application of naturally present foods in contesting several diseases. Meanwhile, country-specific regulations and justification for health claims are some of the challenges herbal nutraceuticals and functional foods market continue to face. Present chapter focused on the nutraceuticals and functional foods with herbs and spices and provides an overview of the herbal nutraceuticals/functional foods market, challenges, motivations, and regulations. This chapter also focused on current status as well as the gaps along with future prospective and developments on nutraceuticals and functional foods with herbs and spices.

Keywords

Functional food; Future prospective; Herbs; Nutraceutical; Regulation; Spices

21.1. Introduction

In recent years, there is an increasing attention in nutraceuticals and functional foods that provide health benefits and are an alternative to modern medicine. Nutrients, herbs (especially herbs and spices), food supplements, and phytochemicals are the main components of nutraceuticals and functional foods that make them essential for maintaining health, acting against various diseases and thus promoting the quality of life. The word nutraceutical is a hybrid of nutrition and pharmaceuticals. It was reportedly coined in 1989 by DeFelice and the Foundation for Innovation in Medicine. Reformulated and clarified in a 1994 press release, its definition was “any substance that can be considered a food or part of a food and that provides health or medical benefits, including the prevention and management of several illnesses.” These products can range from isolated nutrients, diets, herbal products, supplements, and diets to genetically modified designer foods and processed foods such as beverages, grains, and soups [1,2]. According to the International Food Information Council (IFIC), functional foods are “dietary components or foods that may deliver a health advantage beyond basic nutrition.” Health Canada describes functional foods as “similar in appearance to a conventional food, consumed as part of the usual diet, with demonstrated physiological benefits, and/or to reduce the risk of chronic disease beyond basic nutritional functions.” The International Life Sciences Institute of North America has defined functional foods as “foods that by virtue of physiologically active food components provide health benefits afar basic nutrition.” The Nutrition Business Journal classified functional food as “foods fortified with added ingredients or concentrated at functional levels, which improve health or performance. Functional foods include fortified cereals, breads, baby food, bars, sports drinks, ready meals, fortified snacks, and more.” The Nutrition Business Journal claims to use the word nutraceutical for anything that is taken mainly or specifically for health reasons. According to this definition, a functional food would be a kind of nutraceutical. On another side, Health Canada states that nutraceuticals are a product “prepared from food, but sold in the form of pills or powders (potions), or in other medicinal forms that are not generally associated with food” [3,4]. Herbs and spices are used in various forms of nutraceuticals and functional foods. Herbs are plants that have no woody texture and can be treated in many ways depending on individual preferences. Herbs can be dried; however, the drying process leads to a reduction in the effectiveness of the properties of the herbs. Herbs rich in antioxidants have been used for flavor and aroma for over 2000 years [5,6]. Garlic extracts, ginger root, and aloe gel are herbs that have health benefits such as wound healing, reducing cholesterol, antiulcer, and antioxidant activities. Additionally, spices are esoteric food supplements that have been used throughout history to improve the sensual value of food [7]. Spices and herbs, even in trace amounts, have an enormous influence on human health due to their chemopreventive, antioxidant, antiinflammatory, antimutagenic, and immunomodulatory effects on cells and a wide range of beneficial effects on human health. As the potential of spice-derived nutraceuticals against various neurodegenerative diseases becomes more evident, some of these nutraceuticals may be developed as novel nutraceuticals against brain tumors, Parkinson's disease, and other neurodegenerative diseases [8,9].

India and China are home to a large number of medicinal herbs, spices, and tree species, which have a substantially large domestic market without significant foreign competition at the moment. In India, the utmost common forms of nutraceuticals and functional foods are available as traditional Indian Ayurvedic Medicines; these are marketed under different brands. However, it is essential to note that there are no strict pharmaceutical rules and regulations on ayurvedic health products and nutraceuticals in India and China [4]. Additionally, both countries have huge populations, particularly in remote, rural, and inaccessible areas, which are totally dependent on herbal remedies and other naturally available biological resources they use to treat common ailments and as general protective and preventive drugs. In the global marketplace, nutraceuticals and functional foods have become a multibillion-dollar industry, and estimates in Canada indicate that the Canadian nutraceutical and functional foods industry could grow to $50billion (US dollars). Japan is the second largest nutraceutical market in the world after the United States. The nutraceutical market is showing a constant average growth rate of 9.6% per year. The global functional food market was valued at USD 176,518.97 million in 2020 and is expected to grow at a compound annual growth rate (CAGR) of 2.71% over the forecast period 2021–26. According to Precedence Research, the global nutraceutical market will be approximately USD 314.2 billion by 2030, and the global nutraceutical market will grow at a CAGR of 7.5% in 2026. The market was positively obstructed by COVID-19 in 2020. The global pandemic has been shown to affect the proper functioning of herbal functional foods in various regions [10,11]. COVID-19 has also significantly increased consumer interest in herbs and spices, especially nutraceuticals and functional foods, with a significant increase in the number of people expecting functional benefits including immunity from the foods they eat [9,12,13]. The pandemic has created opportunities for many private players in the herbal industries to enter markets to meet the growing demand for nutraceuticals and functional foods, which in turn has benefited emerging growers. In addition, consumers are now ready to experiment with new herbal products, which have prompted companies to look at interesting and affordable products [14].

On the other side, the current state of nutraceuticals and functional foods with herbs and spices in the global market is alarming. The chaotic scene begins with categorization and terminology in which these foods are not considered drugs. There are no uniform, consistent, or standardized rules governing the manufacture, sale, or marketing of these products. All of these products are marketed with strong therapeutic claims as safer natural drug substitutes (Maheshwari et al., 2021). Most of these items can be easily purchased from online retail stores such as eBay, Flipkart, or Amazon without any actual regulation or control. Many sellers are using new transportation technologies as an acquisition strategy to increase sales. In addition, the term “nutraceuticals” is poorly defined worldwide and is not clearly classified as a food or pharmaceutical category from a regulatory perspective. Hence, this is a daunting task for regulators in different parts of the world. However, in the near future, clear and common rules for nutraceuticals and functional foods are urgently required to cope with the rapidly emerging trends and needs in the global marketplace [15]. Therefore, the evidence for the health benefits of nutraceutical and functional foods with herbs and spices in the fields of metabolic health, healthy aging, mental health, and cognition is discussed in this chapter. It's also taken into account to incorporate regulation information of different countries about nutraceutical and functional foods into public health recommendations. The chapter discusses the various ways in which the functionality of herbal-based nutraceutical and functional foods can be considered, laying the groundwork for future studies and cultivating an understanding of herbs and spices' possible contributions to fitness and welfare. In addition, the current status along with future prospective and developments on nutraceuticals and functional foods with herbs and spices is also concise in this chapter.

21.2. Traditional nutraceuticals and products

Food is known to contain nutrients that help nourish our bodies and keep our systems in proper working order. However, since the beginning of human history, it has also been known that certain foods provide additional health benefits, such as preventing and treating several types of diseases. “Let food be medicine and medicine food” is a popular Hippocratic quote that emphasizes the role of food in disease prevention and recognizes the separate role of food, as well as simply the role of the supplier of nutrients (Sohaimy, 2012) [7]. Recently, scientists have focused on the health-promoting effects of medicinal plants/herbs/spices, and there is now abundant evidence supporting the role of various foods/nutritional components in promoting human health [1,9]. Such health-promoting foods or compounds from medicinal plants/herbs/spices have generally fallen into two main categories, namely nutraceuticals and functional foods.

21.2.1. Nutraceuticals

Nutraceuticals are health-promoting compounds or foods that have been isolated or refined from food sources and are usually sold in dosage form (usually tablets, capsules, and liquids) (Fig. 21.1). A good example is a group of compounds called isoflavones, which are isolated from soybeans and packaged in tablets that women can use as an alternative to synthetic compounds during therapy of hormone replacement [9,16]. Other examples of nutraceuticals include herbal extracts, glucosamine, fish oil capsules, chondroitin sulfate tablets, multivitamin tablets containing lutein, and antihypertensive tablets containing peptides derived from fish protein. It is essential to note that the effectiveness of some of these products remains the subject of ongoing research and debate in the scientific community; therefore, consumers should consult their physicians or health-care professionals before using such products, especially if they are already taking medications prescribed by a physician. Herbs and spices play a role in this scenario in a variety of ways [17]. The emphasis of this supplement is on their function in the diet rather than on their use as medications. Identifying specific bioactive compounds to aid in the identification of target benefits will be a part of establishing this position [1]. The list of common herbals with healthy profits as nutraceuticals is presented in Table 21.1 [12].

Figure 21.1
Nutraceuticals (blend of pharmaceuticals and foods).

21.2.2. Functional foods

Functional foods are usually regular foods that are consumed as part of a regular diet, but in addition to providing nutrients, they can also minimize the risk of chronic diseases such as hypertension, cancer, kidney dysfunction, etc. A classic example of a functional food is tomato; this is rich in antioxidant lycopene and reliefs to eliminate toxic compounds from the body and preventing loss to major organs [18,19]. Other examples of functional foods include bran cereals (wheat, rice), soy, oatmeal, fish, and tea (black and green). In addition to traditional foods, there are also functional foods that are made through food processing, such as sour milk, which has antihypertensive effects that have been shown to lower blood pressure (BP) in humans. Different types of functional foods are given in Table 21.2 [17].

With respect to herbal, the functional foods’ approach to herbs and spices could start with how they are used in the meal. For functional foods, there is not any single definition, but the term is used in a variety of ways, such as scientific research, food standards regulation, food marketing, and technical innovation. Functional foods are described as “foods that deliver benefits afar simple nutrition” from a scientific standpoint. However, today's research on food components is expanding beyond principles like preventing clinical deficiency and preserving homeostasis to provide an increasing understanding of how food constituents keenly communicate with the body to promote health and avoid abnormality and overt disease [5,20]. Many clues to this creation can be found in common food consumption across cultures. For example, some traditional Thai food has a cultural history of promoting health through the combination of herbs, spices, and other ingredients, so dietary guidelines should be applied to meals and not to individual foods, as in Western societies (which tend to focus on certain nutrients provided by major food groups). The real difficulty arises when it comes to identifying the benefits and presenting supporting scientific evidence [7,21]. There is ample evidence for the benefits of herbs and spices for cardiovascular and metabolic health, healthy aging, cancer, and mental health. In addition, different ways of looking at the functionality of herbs and spices provide a foundation for future research and contribute to understanding the potential contributions of herbs and spices to health and well-being. Examples of functional foods and their potential health benefits are presented in Table 21.3 [3,22]. The versatility of herbs and spices is demonstrated by considering their nutritional properties from a dietary perspective. The true benefits of including them in the diet, like most foods, are likely to come from a deeper understanding of the health properties that foods best support and from statistical advances in the evidence base for their effects. These advances are well under way, thanks to evidence- based mechanisms to support food-related health claims. Recommendations for the consumption of foods rich in biologically active ingredients such as herbs and spices are now needed. Over time, we should expect more clinical evidence to support the benefits of herbs and spices in maintaining overall health and preventing disease [23,24]. The experimentally proven potential health benefits of the spice are shown in Table 21.4 [9].

Table 21.1

Common herbals with health benefits as Nutraceuticals.

Common name | Botanical name | Active constituent | Health benefits
Garlic | Dried bulbs of Allium sativum (Liliaceae) | Allicin and alliin | Antiinflammatory, antigout, antibacterial, nervine tonic
Ginger | Rhizomes of Zingiber officinale (Zingiberaceae) | Zingiberene and gingerols | Stimulant, hyperglycemia, chronic bronchitis, and throat ache
Maiden hair tree | Leaves of Ginkgo biloba (Ginkgoaceae) | Bilobalide and ginkgolide | PAF antagonist, antioxidant, memory enhancer
Turmeric | Rhizome of Curcuma Longa (zingiberacae) | Curcumin | Anticancer, antiseptic, antiinflammatory, and antiarthritic
Aloes | Dried juice of leaves Aloe barbadensis Mill. (Liliaceae) | Aloesin and aloins | Dilates capillaries, emollient, antiinflammatory, wound-healing properties
Ginseng | Dried root of Panax ginseng (araliaceae) | Panaxosides and ginsenosides | Stimulating nervous and immune system and adaptogenic properties
Liquorice | Dried root of Glycyrrhiza glabra (leguminosae) | Liquirtin and glycyrrhizin | Antiallergic and antiinflammatory, expectorant
Senna | Dried leaves of Cassia angustifolia (Leguminosae) | Sennosides | Purgative
Onion | Dried bulb of Allium cepa Linn. (Liliaceae) | Alliin and allicin | Hypoglycemic activity, antiatherosclerosis, and antibiotic
Brahmi | Herbs of Centella asiatica (Umbelliferae) | Madecassoside and asiaticoside | Nervine tonic, antianxiety, spasmolytic
Echinacea | Dried herb of Echinacea purpurea (Asteraceae) | Echinacoside and alkylamide | Antiinflammatory, antiviral, immunomodulator
Bael | Unripe fruits of Aeglemarmelos Corr. (Rutaceae) | Marmelosin | Digestive, treatment of diarrhea, appetizer, and dysentery
Asafoetida | Oleo gum resin of ferula assafoetida L. (umbelliferae) | Umbellic acid and ferulic acid | Stimulant, expectorant, carminative
Valeriana | Dried root of Valeriana officinalis Linn. (Valerianaceae) | Valerate and valerenic acid | Tranquillizer, intestinal cramps, bronchial spasm, menstrual and migraine pain.
Goldenseal | Dried root of Hydrastis canadensis. (Ranunculaceae) | Berberine and hydrastine | Antimicrobial, treatment of mucosal inflammation, astringent, antihemorrhagic

Adapted from B. Chauhan, G. Kumar, N. Kalam, S.H. Ansari, Current concepts and prospects of herbal nutraceutical: a review, J. Adv. Pharm. Technol. Res. 4 (1) (2013) 4.

Table 21.2

Different types of functional foods.

Type | Description | Example
Enriched food products | Adding new ingredients or nutrients that are not usually found in food | Fruit juice enriched with calcium, foods with prebiotics and probiotics
Fortified food products | Increase the content of nutrients present | Grain products fortified with folic acid, fruit juice with extra VC
Enhanced food commodities | Changes in raw materials that altered the nutrient composition | Carotenoid-containing high-lysine corn, potatoes
Altered food products | Change existing components with beneficial components | Low-fat foods with fat replacers

Adapted from A.T. Khalaf, Y. Wei, S.J. Alneamah, S.G. Al-Shawi, S.Y. Kadir, J. Zainol, X. Liu, What is new in the preventive and therapeutic role of dairy products as nutraceuticals and functional foods? BioMed Res. Int. (2021) 8823222.

Table 21.3

Examples of functional foods and their potential health benefit.

Functional foods | Potential health benefits
Whole foods
Vegetables and fruits | Reduced risk of heart diseases and cancers
Flaxseed | Reduced risk of certain cancers and heart diseases, increases blood glucose control, reduces triglycerides
Garlic | Reduced risk of cancers and heart diseases, reduces cholesterol
Soybean | Regulates menopause systems, reduced cholesterol, prevents osteoporosis and heart diseases
Fish | Reduces triglycerides and cholesterol, reduced risk of heart diseases
Enhanced foods
Fish oil with omega-3 fatty acids | Reduced risk of heart diseases
Dairy products with probiotics | Controls eczema and diarrheal disorders, reduced risk of colon cancer
Fortified foods
Grains with folic acid | Reduced risk of heart diseases and neural tube defects
Milk with vitamin D | Reduces risk of osteoporosis and osteomalacia
Juices with calcium | Reduce risk of hypertension and osteoporosis
Grains with added fiber | Increase blood glucose control, reduced risk of heart diseases and cancers, reduce constipation and cholesterol

Adapted from K. Gul, A.K. Singh, R. Jabeen, Nutraceuticals and functional foods: the foods for the future world, Crit. Rev. Food Sci. Nutr. 56 (16) (2016) 2617–2627.

Table 21.4

Experimental documented potential health benefit of some spices.

Potential health benefits | Spices observed to exert
Hypoglycemic potential | Turmeric, garlic, fenugreek, onion, cumin
Cancer/antimutagenic effect preventing | Curcumin/turmeric, mustard, gingerol/ginger
Antioxidant effect | Curcumin/turmeric, eugenol, capsaicin
Prevention and dissolution of cholesterol gallstones | Capsaicin, curcumin
Antidepressant | Cloves, ginger, black pepper, all-spice
Lowering of blood cholesterol | Red pepper/capsaicin, garlic, fenugreek, onion, curcumin/turmeric
Antiarthritic effect and antiinflammatory | Curcumin/turmeric, eugenol, capsaicin
Amelioration of diabetic nephropathy | Onion, curcumin
Meningitis | Garlic
Antimicrobial | Curcumin/turmeric, aasafoetida, garlic
Protection of erythrocyte integrity in hypercholesterol-emic condition | Garlic, curcumin, capsaicin
Schizophrenia | Onion
Digestive stimulant action | Coriander, capsaicin, aajowan, piperine, ginger, curcumin, onion, cumin, fennel, mint
Spongiform encephalopathy | Turmeric

Adapted from E.B. Daliri, B.H. Lee, Current trends and future perspectives on functional foods and nutraceuticals. In Beneficial Microorganisms in Food and Nutraceuticals, Springer, Cham, 2015, pp. 221–244.

21.2.3. Difference between nutraceuticals and functional foods

The key difference between nutraceuticals and functional foods is that nutraceuticals are bioactive compounds found in fortified foods, nutritional supplements, or herbal products, while functional foods are foods with bioactive compounds such as beta-carotene, ferulic acid, resveratrol, lycopene, and many others (Fig. 21.2). In addition, functional foods contain naturally occurring bioactive compounds, and nutraceuticals can be in the form of capsules, pills, or liquids. Today, the use and study of disease-fighting properties of a variety of phytochemicals found in both nonfood and edible plants have led to a resurgence of research in human health and nutrition [25,26]. The difference between nutraceuticals and functional foods is summarized in Table 21.5 [17].

Figure 21.2
Difference between nutraceuticals and functional foods.

Table 21.5

Difference between nutraceuticals and functional foods.

Nutraceuticals | Functional foods

The foodstuff (as a fortified food or a dietary supplement) held to provide health or medical benefits in addition to its basic nutritional value

The foods with ingredients which give that food health-promoting properties over and above its usual nutritional value

Natural or synthetic and available as pills, capsules, or liquids | Natural
The bioactive compounds found in fortified foods, dietary supplement, or herbal products | Naturally contain bioactive compounds found in foods
Include traditional nutrients | The bioactive compounds in them are different from the traditional nutrients
Beta-carotene, resveratrol, ferulic acid, and lycopene are examples | Grapes, apple, and strawberries are examples

Adapted from A.T. Khalaf, Y. Wei, S.J. Alneamah, S.G. Al-Shawi, S.Y. Kadir, J. Zainol, X. Liu, What is new in the preventive and therapeutic role of dairy products as nutraceuticals and functional foods? BioMed Res. Int. (2021) 8823222.

21.3. Health benefits of nutraceuticals and functional foods

Human health and well-being are largely determined by the consumption of nutritious food. The emergence of products that are beneficial to health provides an excellent opportunity to improve the health of the population; therefore, in recent years, these compounds have attracted a lot of attention from the scientific community, food manufacturers, and consumers. Several epidemiological studies over the past 50 years have clearly shown that diets dominated by fruits, vegetables, and dietary fiber (plant/herbal foods) prevent and reduce the risk of chronic diseases (e.g., cardiovascular disease, obesity, diabetes, etc.), and they also strengthen human health. A generation of scientific research around the world linking plant foods to health has led to the recognition that bioactive compounds in plants have health benefits [23,27 ]. High dietary intake of plant extracts, herbs, spices, phytochemicals, fruits, vegetables, and whole grains is closely associated with a reduced risk of chronic diseases such as cancer and cardiovascular disease, which are the leading causes of death in Europe and the United States. It is projected that one-third of all cancer deaths in industrialized nations can be prevented with adequate dietary formula. This suggests that dietary behavioral changes, such as increased intake of plant extracts, herbs, spices, phytochemicals, fruits, vegetables, and whole grains, and associated lifestyle changes, are practical strategies for reducing significant cancer incidence [12,28]. There is extensive scientific literature that links the consumption of diets rich in herbs and spices with a reduced risk of cancer, especially epithelial cancer of the digestive and respiratory systems. According to the results of 206 epidemiological studies in humans and 22 studies in animals, similar effects of herbal diets correspond to cancers of the stomach, esophagus, lungs, oral cavity and pharynx, endometrium, pancreas, and colon. The types of vegetables and fruits that most often protect against cancer are raw vegetables, followed by onions, carrots, herbs, crucifers, and tomatoes. These include, but are not limited to, the treatment of cancer, atherosclerosis and cardiovascular disease, the aging process, enhancing the immune response, and diabetes [3,29].

21.4. The global nutraceuticals and functional foods market

The global market for nutraceuticals and functional foods has been categorized by type, benefit, ingredient, origin, and consumer. Foods, supplements, and beverages are classifications of types of nutraceuticals and functional foods. Based on the advantages of this market, it has been divided into health and disease prevention, wellness, beauty, fitness, and other benefits. Nutraceuticals and functional foods are classified as animal or plant based on origin. The key ingredients of these goods are phytochemicals, medicinal plants, herbs, spices, carotenoids, minerals, fatty acids, and other vitamins. End users are divided into the elderly, adults, and children [12,30]. The growth (in billions of dollars) of the global nutraceutical and functional foods market is shown in Fig. 21.3 [10,11]. The main factors driving the market are the rapid growth of food science and technology, people's interest in well-being through diet planning, rising health-care costs, and an increasing aging population. The entire market was also segmented by region: America, Asia-Pacific (APAC), Europe, and Rest of the World. The data show that America will become the market leader in functional foods and nutraceuticals in the near future, followed by APAC and Europe. These regions are also encouraging consumers to root out the growing health problems in the market. APAC enters the market with the packaged food industry and new growth opportunities in the food industry. As per precedence research, the size of the global nutraceutical market is expected to be about $314.2 billion by 2030, and the global nutraceutical market will grow at a CAGR of 7.5% over the forecast period (2021–26). The global functional food market was valued at $176,518.97 million in 2020, and the market is expected to grow at a CAGR of 2.71% over the forecast period 2021–26 [10,11]. The main factors contributing to the growth of functional foods are: the trend toward rapidly resolving health problems, rising health-care costs, physical inactivity, challenging lifestyles, heightened health awareness, ease of consumption, prevention rather than response to health problems, and increasing aging of the population. Quantitative and qualitative data for the global market of nutraceutical and functional foods are obtained from various primary and secondary bases. Number of questionnaires and interviews were conducted with vice presidents, executives, and deputy managers associated with the various companies that manufactured these products [9]. Extensive research was conducted to cover all key developments and technology trends for market analysis. The data were collected by the Association of Agricultural Equipment Manufacturers, the Association of Agricultural Engineers, the Association of Equipment Manufacturers, etc. [14].

Figure 21.3
The growth (in USD billion) of global nutraceuticals and functional foods market.

21.5. Motivators of nutraceutical and functional foods market

Consumer awareness in the relationship between diet and health has skyrocketed around the world. It is much more widely recognized today that people can reduce their risk of disease and maintain health and well-being through healthy living and diet. Continuous support for the main role of foods such as fruits, vegetables, and whole grains in disease prevention, as well as dietary antioxidants and combinations of protective substances such as plant extracts, herbs, and spices have helped stimulate new developments in functional aspects in the global food market [29,31]. Changes in socioeconomic and demographic shifts in the population have increased the need for herbal foods with additional health benefits. As per a report from the US Department of State and the United Nations Department of Economic and Social Affairs, approximately 8% of the world's people are over 65 (Fig. 21.4), and this age group is expected to be around 13% of the population by 2030, and most of them live in developing countries [9,32]. Thus, an increase in life expectancy necessitates an improvement in the quality of life. In addition, rising health-care costs have prompted governments, researchers, health-care professionals, and the food industry to look for ways to effectively manage these changes [33,34]. These are important determinants of the functional food market. The popularity of nutraceuticals/functional foods with herbs and spices is due to rising rates of diabetes and obesity, rising eye health problems, rising rates of cardiovascular disease, and growing preference for preventive medicine. Other ingredients such as multivitamins, omega-3s, and astaxanthin are the other driving forces behind the growth of nutraceutical and functional food market. Herbal and spice-based nutraceuticals are gaining popularity, as are functional foods that require weight management, immunity, cardiac health, and digestive health [35]. Product innovations have also propelled consumers to promote functional foods and nutraceuticals. Consumers are looking for convenience as well as products with greater bioavailability of the main ingredients [9,36].

Figure 21.4
Percent of world population over 65 years according to US state department and united nations department of economic and social affairs.

21.6. Present status of nutraceutical and functional foods

User interest in the relationship between health and diet has improved the demand of nutraceuticals and functional foods with herbs and spices. Fast developments in technology and science, rising health-care costs, an aging population, and changes in food laws affecting product labeling and indications, and a growing interest in achieving wellness with herbs and spices are factors that are increasing interest in nutraceuticals and functional foods. Reliable scientific studies point many possible health benefits of herbs and spices [7,37]. The growth in demand for herbal nutraceuticals and functional foods around the world is driven by several factors, including the following.

21.6.1. Affluence

Population growth with lifestyle changes due to increased labor force participation, sedentary lifestyles, an increase in the number of working women, changing dietary habits, and higher disposable income indicates a demand for herbal and spices origin nutraceuticals and functional foods. The population of young people is growing rapidly, with completely different nutritional and nutritional needs. Likewise, the number of older adults who need to introduce certain specialized herbal foods is also increasing [38].

21.6.2. Affordability

More than 50% of household spending goes on food alone, so with additional disposable income from rising per capita incomes, consumers may be spending on nutraceuticals and functional foods containing herbs and spices. In addition, out-of-pocket spending, which accounts for 64% of health-care spending, and the rapidly rising cost of health care are also driving consumers to seek healthy and functional foods.

21.6.3. Awareness

The popularization of new natural herbal products and nutraceuticals through print, electronics, and other media has led to an increased awareness of the general public about the health problems associated with diet and the possible health components present in them. Doctors have begun prescribing herbal functional foods/nutraceuticals in addition to conventional medications. In addition, the availability of diagnostic equipment at an affordable cost allows people to monitor important health parameters such as BP, lipid profile, fasting glucose, etc., to ensure good health and take corrective action if desired.

21.6.4. Accessibility

More and more functional foods and nutraceuticals from herbs and spices are appearing on the market day by day. Every year, hundreds or many nutraceuticals and functional products are introduced around the world, using various herbs and spices as their main components. The growth of the retail sector has further increased the availability of these products to consumers. Many multinational companies have opened their stores to sell these herbal nutraceuticals and functional foods. Depending on basic needs, these herbal products can be divided into three parts.

21.6.4.1. Foundation needs

To support normal growth and well-being, these herbal products contribute to overall well-being and healthy lifestyles. Products include raw materials fortified with macro- and micronutrients such as herbal extracts, raw herbs, chyawanprash, phytochemicals, processed spices, juices, flour, and probiotic products.

21.6.4.2. Condition-specific needs

This requires nutraceuticals/functional foods under certain conditions, especially when they are in short supply. The segment includes sick, elderly, pregnant, lactating, lactating people, etc. Most of the population suffering from disorders associated with micronutrient or macronutrient deficiencies, obesity, heart and osteoporotic diseases may require special herbal nutraceuticals and functional foods. The market for such goods is estimated to grow to $50–99.5billion from the present value of $10billion.

21.6.4.3. Enhancement needs

These herbs as well as spices nutraceuticals and functional foods are designed to meet the increased nutritional needs of certain groups, including athletes. Foods include high energy powders, electrolytes, protein powders or drinks, amino acids, and other ergogenic supplements. The market is in its infancy and is estimated to reach $1.5billion.

21.7. Challenges in the nutraceuticals and functional foods market

Although the market for nutraceuticals and functional foods has been around for several years, but its future is difficult to predict due to issues such as country regulations, difficulties in justifying health claims, and lack of innovation from food companies. The world has seen mixed acceptance of nutraceuticals and functional foods with herbs and spices. Although several countries have legislation allowing the use and regulation of health claims, the process has not resulted in the authorization for the use of these claims. Validation of health claims for functional foods is difficult in the United States and Japan. Although the European Food Safety Authority (EFSA) has offered guidelines for justifying claims of health benefits in the European community, this still seems impossible there. The type of indication and factors such as period of clinical trials, validation of biological markers, dose-response curves for defining effective dose, and side effects that would support a health claim are not defined in these regions. In addition, for companies that are considering a health claim petition and get approval, the competing companies can use the claim. While functional products and nutraceuticals with exclusive ingredients can be patented, the vast majorities of products contain “free” ingredients and therefore can be easily copied [32]. This situation gives the parent company a slight competitive advantage. Again, as functional foods and nutraceuticals are turning to health care, health claims must be supported by scientific evidence and this is not an easy assignment. Biomarkers for improved health or reduced risk are not easy to recognize. In addition, clinical trials are often time-consuming and provide little conclusive evidence for dosing efficacy, while side effects often require follow-up studies. Aside from the difficulty of validating health claims, finding a good product is not easy [39,40]. Moreover, the nutraceutical industry continues to exist more as a concept than a reality because it includes nutritional supplements and functional foods. The immense majority of food houses are not very innovative because they simply “functionalize” conventional foods by adding some vitamins, minerals, or herbal extracts, rather than creating new food concepts of ingredients that have overall health benefits [7,41].

21.8. Regulatory and perception issues

Nutraceuticals and functional foods are controlled as food or dietary supplements, depending on how they are branded. Nutraceuticals and functional foods may contain health claims, usually preapproved by the Food and Drug Administration (FDA), or structure/function statements based on the most recent regulations issued by the FDA in accordance with the Dietary Supplement Health and Education Act (DSHEA) 1994. The FDA vaguely regulates medical food products within its own regulatory category. Health claims define the relationship between diet and disease, and only 11 health claims are approved by the FDA [42]. Example of this is “a healthy diet with adequate folate can reduce a woman's risk of having a baby with a brain or spinal cord weakness.” Health claims have traditionally been based on the concept of “significant scientific agreement,” which the FDA recently defined in a guidance document as an agreement between qualified scientific experts that there a substance/disease relationship occurs based on a sound form of technical evidence. Evidence must be strong enough so that it cannot be reversed in subsequent studies [12,43,44]. Scientific associations such as the Institute of Food Technologists of Chicago and the American Dietetic Association in Washington, DC support the potential profits of functional foods when supported by scientific evidence. However, a leading customer organization, the Center for Science in the Public Interest (CSPI) in Washington, DC, recently released a report criticizing functional foods. This has been driven by the growing market entry of many functional foods that are high in fat, calories, and sodium. The CSPI noted that “unless governments require functional ingredients to be proven to be effective (and safe) before they are added to food, unless properly informed claims are required, if functional ingredients are simply added to foods high in fat, cholesterol, sodium, or sugar, so dubious functional foods may just be 21st century quackery” [26,42,44].

21.9. A snapshot of global regulations

21.9.1. United States

In the United States, nutraceuticals and functional foods are primarily categorized as vitamins and nutritional supplements. A number of regulatory bodies are constituted there for the regulation of such products. One of them is the DSHEA 1994, which deals with food additives. Another one is the Federal Food, Drug, and Cosmetic Act, which applies to all food and nutritional supplements. Last one is Good Manufacturing Practice regulations since 2007, which regulates manufacturing process. As per these guidelines, the manufacturer: is accountable for ensuring the safety of the additives whatever they manufacture or distribute in market, must not make wrong or confusing claims, cannot claim that the product will diagnose, treat, alleviate, cure, or prevent a disease, must promise the purity, concentration, identity, and composition of its supplement (the deadline for adherence by all manufacturers is June 2010). In 1988, the FDA granted medical food to “orphan drug” status to stimulate development, and some US health plans are reimbursing the cost of medical products [26,45].

21.9.2. European Union

In the European Union (EU), nutraceuticals and functional foods are subject to a complex set of rules, depending on their type. They are mainly controlled by the EFSA, which also evaluates claims and maintains a public register of health and nutrition claims. If a product is not on this certified list, it should not be used until it has been approved for inclusion for extensive scientific testing by the manufacturer and subsequent evaluation by EFSA. The EFSA also fixed minimum and maximum concentrations for constituents to be added to food additives. The labels of the products cannot claim that it will help to mitigate, diagnose, cure, or prevent disease, while claims-related nutrition and health must be approved at EU level. Between 2011 and 2008, EFSA assessed 2758 food-related health claims to ensure they were scientifically supported. About 10% of claims were substantiated only [26,45].

21.9.3. India

The production, packing, supply, sale, storage, and import of nutraceuticals and functional foods in India are controlled by the Food Safety and Standards Act. The necessary manufacturing oversight is less stringent than pharmaceutical oversight, which could affect the capability of Indian manufacturers to production nutraceuticals and functional foods for export [45].

21.9.4. Japan

In Japan, foods with nutritional requirements are not strictly regulated and simply must meet minimum and maximum daily concentrations of five minerals and 12 vitamins. In Japan, the regulatory body “Food for Specified Health Uses” needs approval before being placed on the market and refers to foods containing dietary ingredients that have positive physiological effects and promote health. No indication of disease risk reduction is permitted in Japan [26,45].

21.9.5. China

In China, the registration procedure is long and expensive, and such food industries are regulated by three main regulatory bodies. The State Food and Drug Administration (SFDA) is responsible for the registration and dietary supplements. The Ministry of Health supervises the SFDA and gives approval of new food materials. Lastly, the Administration of Quality Supervision Inspection and Quarantine regulates all exports and imports [26,45].

21.10. Toxicity potential of nutraceuticals and functional foods

Many people trust that nutraceuticals and functional foods, especially medicinal plants/herbs/spices, are significant preparations for solving health problems without side effects. This trust stems from the fact that they have been used for a long time without serious toxic effects, while this is true of a wide variety of herbal nutraceuticals/functional foods that have fewer side effects than pharmaceuticals. In addition, conventional medicine believes that if a drug is effective, it will inevitably have toxic or side effects. Medical institutions believe that medicinal herbs are drugs and therefore must have side effects [46,47]. So, they must be prepared with the accurate constituents and used with care. People use many of different types of herbs and spices as nutraceutical/functional foods to fulfill their basic need of nutrition, but only a limited number of them have undergone serious safety studies. Many of them remain poorly studied and mostly underdeveloped, and their wild relatives are endangered and in need of protection. The management of these valuable plant resources will require rigorous scientific research combined with an approach that respects and values traditional knowledge systems [48,49].

21.11. Future prospects of nutraceuticals and functional foods

Nutraceuticals/functional foods products derived from herbs and spices have been accepted for having several biological characteristics that contribute to their positive properties on the health of human being. Classes of phytochemicals such as alkaloids, saponins, glycosides, flavonoid, steroids, stannins, and terpenoids are often isolated from various plant parts that play an important role in maintaining human health, thus can be used in nutraceuticals as well as functional foods. Nutraceuticals/functional foods derived from herbs and spices could be further used to treat chronic disease, enhance human health, and care different types of metabolic disorders [45,50]. Such kind of nutraceutical/functional food research is attracting more and more attention to therapeutic effects, especially in the time of a pandemic. The manufacturers of supplement show interest in substituting the synthetic constituent with natural plant substances due to the high demand for natural products. Companies of nutraceuticals and functional foods, especially herbal origin, are well aware of production costs and margins. Nutraceutical and functional foods manufacturers are also frequently launching new herbal products to the market to grow nutraceutical and functional food industries [7]; (Maheshwari et al., 2021). In addition, the use of nutraceuticals for the management and inhibition of chronic illnesses is encouraged. For example, to prevent cancer, different types of green tea and soy foods are used. Furthermore, increased research into the safety and efficacy of these new products is driving increased investment in applications and technology. Emerging molecular biology technologies such as nutrigenomics, nutriproteomics, and nutrigenetics are highly recommended in nutraceutical research. Innovations in plant technology can also play a role in further advances in the development of foods that improve human health [4,51]. Furthermore, the identification of single nucleotide polymorphisms in the human population has made it possible to predict variations in individual drug responses and materialize a new concept of personalized drugs. Consequently, nutritional genomics emerged, which comprises the interaction of dietary components with genomes and determines proteomic and metabolic changes, called “nutrigenomics.” In addition, an understanding of the genetic differences between humans has developed; people react differently to the same nutraceutical called nutrigenetics. The accessibility of genomic information is accelerating the treatment of disease, and pharmaceutical industries are using genotyping to envisage the efficacy, toxicity, and safety of preparations throughout clinical trials. Pharmacogenomics studies a patient's response to medicines, while “nutrigenomics” was designed to study the effects of nutraceuticals and dietary components on the health of an individual. Nutrigenomics uses the genetic information of an individual to predict the intake of nutraceuticals to keep health and inhibit disease [1,9]. In the meantime, government funding of clinical trials has increased and supports have also increased for research on herbals/plants/phytochemicals-based functional foods and nutraceuticals. Regulatory authorities regulate the supply of nutraceuticals/functional foods and also prepare systematic analytical monographs for routine quality control for these types of nutraceuticals/functional foods. The analytical profiles of these products are published by consumer organizations, and it enables the customer to select the highest quality goods. The available data related to nutraceuticals and functional foods against certain life-threatening illnesses are still not enough to exhibit the use of nutraceuticals/functional foods from herbs and spices [49,52]. Consequently, government support is imperative to increase or reinforce research in these fields.

21.12. Nutraceuticals/functional foods: the future of intelligent food

Diseases due to poor and imbalance lifestyle potentially costing the National Health Service more than £10billion a year in the United Kingdom. There is a huge incentive for governments to focus on preventive care and consumer awareness of their health. Food and pharmaceutical companies are well positioned to generate profits. Food companies have more knowledge than pharmaceutical nutrition and food formulation, and decades of customer research and affairs with mass market traders, but their research experience is not as extensive as in the pharmaceutical industry. Nutraceutical food cycles are much longer than food cycles, which should reflect on the strengths of the pharmaceutical industry. However, pharmaceutical companies face a different kind of challenges [1,45,50]. They have well-established research industries, regulatory professionals, and shareholder relationships that empower them to create new molecules, as well as broader connections with the scientific community. However, they have great resources for sales and marketing, but most of their efforts are intended at health-care and pharmaceutical professionals rather than customers. Only a few numbers of pharmaceutical industries have a customer arm. By bringing together the strengths (and weaknesses) of each, a merger of the nutraceutical portfolios of a large food company and a large pharmaceutical company might make sense, but not much has happened yet. Instead, the line between the two industries and an active market for corporate transactions, involving relatively small acquisitions and liquidations, was blurred: preliminary skirmishes in a long-term battle [7,9].

21.13. Characteristics of success: daring to win

21.13.1. Technology

Technology will be critical. The market for nutraceuticals and functional foods with herbs and spices is probable to profit from the personalized medicine trend, and the development of technologies to facilitate this movement is critical, including through the huge amount of data that will drive more and more health-care connected system. Consumer companies have the intelligence to personalize dietary advice as well as with smart partnerships, pharmaceutical companies can tailor over-the-counter nutraceuticals to fit consumers' lifestyles [41,45].

21.13.2. Compliant marketing

The market for herbal nutraceuticals and functional foods is heterogeneous globally: countries differ in how their regulatory bodies distinguish nutraceuticals as pharmaceuticals rather than food, while customers themselves cannot agree on which category of these items are best suited. Successful manufacturer will have to grow a detailed global marketing campaign that can exploit economies of scale by acclimating to national preferences [45].

21.13.3. Regulation

The rules and regulations for nutraceutical/functional foods based on herbs and spices vary from country to country and are constantly changing. In the EU, products that claim to be nutraceutical/functional foods must be certified by the European Food Standards Authority, and the same regulations work in Canada too. But this contrasts with the Japan and the United States. Markets of these countries currently account for more than half of the world's demand for herbal nutraceuticals and functional foods, where products do not have to clear rigorous government parameters until they pass to cure or prevent a certain disease. Manufacturers of these types of products will need to work closely with governments to anticipate market opportunities that might arise when regulations change [41,45].

21.13.4. Supply chain

With growing customer worries related to the safety of foods, the quality of the herbs and spices used to produce nutraceutical and functional foods has received close scrutiny, so companies need to carefully consider how to access and supply key constituents or flavors. Several recent attainments of food and beverage manufacturers aim to ensure a stable and high-quality supply of raw materials for these herbal products. For example, vertically integrating supply chains provide greater assurance of supply and set the stage for developing new uses for existing materials or for producing new related ingredients [41,45].

21.13.5. Product strategy

There may be few blockbusters in this industry, and some industries have recently rebalanced their product portfolios by ditching product lines that do not align with their policies. Companies with a superior product strategy will gain a competitive edge: combining ingredients, technologies, and labeling to adjust product portfolios in every country [45,53].

21.13.6. Deal making

Herbal and ayurvedic manufacturers can compensate for their shortcomings in certain ranges by being rapid and critical in business negotiations. It will only take a large acquisition to change the dynamics of the industry, and other large manufacturers in this field will be forced to respond. Well-established pharmaceutical and herbal companies are not known for being entrepreneurial, so they will have to do many things that sometimes go beside their corporate principles such as: be bold, sometime flexible, and tolerant at the same time [53]. Ayurvedic, herbal, and pharmaceutical manufacturers will have their competitive edge in the battle for the market for functional foods and nutraceuticals based on medicinal and herbal plants. The champions are likely to be those performers who can have a long-term vision, as they skillfully and decisively seize short-term opportunities as they arise [41,45].

21.14. Conclusion

The lifestyle of people in the modern era has faced many challenges, especially many health problems such as diabetes, obesity, cancer, osteoporosis, and various chronic intestinal problems. These problems in life can develop from eating more unhealthy food, which leads to health issues and is caused by nutritional deficiencies in the food. Due to the increased awareness of people about food, synthetic food ingredients have been replaced with nutraceutical/functional foods from herbs and spices origin. Global consumption of nutraceuticals and functional herbal foods is growing rapidly due to their perceived stability, accuracy, safety, and efficacy. However, these nutraceutical/functional products pose many challenges with regard to their effectiveness, purity, and regulatory requirements. In addition, it is challenging to determine the corresponding biologically active substances and to establish their role in the fight or treatment of various diseases. More research is needed to confirm the possible health benefits of these plant foods, for which the relationship between diet and health has not been scientifically proven enough. No regulatory body has been established to regulate, standardize, and control herbal nutraceuticals and functional foods across the world. Hence, stringent and internationally agreed guidelines will be required to ensure that adequate information is used to support any health claims on product labeling and will be detailed in the safety recommendations/guides to be incorporated with such type of marketing items. There is a vast gap of knowledge existing on the part of both the producers and customers. Because of ancient valuable knowledge along with progressive research on nutraceuticals/functional foods with herbs and spices, their future may be assumed to be bright in the world in coming years.

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46. Nicoletti M. Nutraceuticals and botanicals: overview and perspectives.
Int. J. Food Sci. Nutr.
2012;63(Suppl. 1):2–6.

47. Subedi L, Tchen S, Gaire B.P, Hu B, Hu K. Adjunctive nutraceutical therapies for COVID-19.
Int. J. Mol. Sci.
2021;22(4):1963.

48. Jędrusek G.A, Górecka D, Buchowski M, Wieczorowska‐Tobis K, Gramza‐Michałowska A, Szymandera‐Buszka K. Recent progress in the use of functional foods for older adults: a narrative review.
Compr. Rev. Food Sci. Food Saf.
2020;19(2):835–856.

49. Muhammad D.R, Dewettinck K. Cinnamon and its derivatives as potential ingredient in functional food—a review.
Int. J. Food Prop.
2017;20(Suppl. 2):2237–2263.

50. Escalante F.M, Pérez‐Rico D.A. Advancements in algae in nutraceutical and functional food. Recent Advances in Micro and Macroalgal Processing: Food and Health Perspectives. . 1. 2021:506–536.

51. Birch C.S, Bonwick G.A. Ensuring the future of functional foods.
Int. J. Food Sci. Technol.
2019;54(5):1467–1485.

52. Misra S, Pandey P, Mishra H.N. Novel approaches for co-encapsulation of probiotic bacteria with bioactive compounds, their health benefits and functional food product development: a review.
Trends Food Sci. Technol.
2021;109:340–351.

53. Reque P.M, Brandelli A. Encapsulation of probiotics and nutraceuticals: applications in functional food industry.
Trends Food Sci. Technol.
2021;114:1–10.

54. Kandylis P.
Grapes and Their Derivatives in Functional Foods
. 2021.

Further reading

[1]. Granato D, Barba F.J, BursaćKovačević D, Lorenzo J.M, Cruz A.G, Putnik P. Functional foods: product development, technological trends, efficacy testing, and safety.
Annu. Rev. Food Sci. Technol.
2020;11:93–118.

[2]. Maheshwari R, Kuche K.N, Advankar A, Soni N, Raval N, Sharma P.A, Tekade M, Tekade R.K.
Natural Ingredients/Botanical Extracts for the Nutraceutical Industry. In Flavors for Nutraceutical and Functional Foods
. CRC Press; 2018:75–122.

Index

Note: Page numbers followed by ‘f’ indicate figures those followed by ‘t’ indicate tables.

A

Acetylcholinesterase (AChE) inhibition, galangal, 103

Advanced glycation end products (AGEs)

Boswellia serrata gum resin extract, 37

cinnamon, 268

Aglycones, 112–113
,
112f

Air pollution, Ocimum sanctum, 64
Ajasrika, 4
Alkaloid, fenugreek, 135–136
Alpinia officinarum, 95
Alzheimer’s disease

cinnamon, 267

curcumin, 22

Ocimum sanctum, 63–64

Analgesic activity, Myristica fragrans fruit, 298

Animal Product Act 1999, 339–340
Anthelmintic activity, fenugreek, 141–142
Anthocyanins, Garcinia cambogia, 214–215
Antiacnes activity, pomegranate, 189

Antiadiposity properties, Garcinia cambogia, 214–215

Antiaging effect

asafoetida, 239

Myristica fragrans fruit, 300

Antibacterial activity

Camellia sinensis (tea), 223

cinnamon, 265–266

fenugreek, 140

pomegranate, 185

Anticancer activity

asafoetida, 234–235

Boswellia serrata gum resin extract, 36

Camellia sinensis (tea), 222–223

cinnamon, 266–267

fenugreek, 137–138

galangal, 100–101

Moringa oleifera, 80–81

Myristica fragrans fruit, 299

piper species and P. nigrum, 46–47
,
48t–50t

pomegranate, 181–182

ursolic acid (UA), 162

Anticoccidial activity, pomegranate, 186
Anticonvulsant effects, asafoetida, 236
Antidiabetic activity

asafoetida, 238

Asparagus racemosus, 250

cinnamon, 263–264

fenugreek, 138

Moringa oleifera, 81–82

Myristica fragrans fruit, 297

pomegranate, 188–189

ursolic acid (UA), 163

Antidiarrheal effects, Asparagus racemosus, 248–249

Antiepileptic effects, Asparagus racemosus, 250

Antifertility potential, Ocimum sanctum, 65

Antifungal activity, fenugreek, 140

Antihyperglycemic activity, Moringa oleifera, 82

Antihypertensive activity, Camellia sinensis (tea), 225

Antiinflammatory activity

asafoetida, 236

cinnamon, 265

galangal, 101

Moringa oleifera, 77–78

Myristica fragrans fruit, 298

piper species and P. nigrum, 47

pomegranate, 186–188

Antileishmanial activity, pomegranate, 186

Antilice and antidandruff activity, pomegranate, 189

Antimicrobial activity

asafoetida, 237–238

Asparagus racemosus, 249

Boswellia serrata gum resin extract, 37

Camellia sinensis (tea), 224–225

galangal, 103–104

Moringa oleifera, 75–77

Myristica fragrans fruit, 298–299

piper species and P. nigrum, 47–51

ursolic acid (UA), 162

Antinociceptive effects, asafoetida, 236

Antiobesity activity

asafoetida, 238

fenugreek, 139

galangal, 101–102

Garcinia cambogia, 215

Myristica fragrans fruit, 300

piper species and P. nigrum, 51

ursolic acid (UA), 163

Antioxidant activity

Boswellia serrata gum resin extract, 37

Camellia sinensis (tea), 221–222

cinnamon, 262–263

fenugreek, 141

galangal, 102

Moringa oleifera, 78–79

Myristica fragrans fruit, 298

Ocimum sanctum, 64–65

piper species and P. nigrum, 45–46

pomegranate, 183–185

Antiparasitic activity, asafoetida, 238
Antispasmodic therapy, asafoetida, 236–237
Antistress effects, Ocimum sanctum, 64
Antiulcer effects

Asparagus racemosus, 248–249

cinnamon, 267

fenugreek, 142

Antiviral activity

Camellia sinensis (tea), 223–224

galangal, 103

pomegranate, 185–186

Anxiolytic effect, asafoetida, 236
Arjuna Ksheerpaka, 4
Asafoetida

antiaging effect, 239

anticancer activity, 234–235

antidiabetic and antiobesity effects, 238

antimicrobial activity, 237–238

antiparasitic activity, 238

blood and blood vessels, 239

chemical constituents, 233–234
,
234t

exudates, 233

fractions, 234t

gastrointestinal tract, 236

hepatoprotective properties, 237

metabolic activity, 239

neuroprotective effects, 235–236

odor, 233

relaxant and antispasmodic effects, 236–237

reproductive effects, 239

toxic effects, 239–240

Ashtawarga plants

ayurvedic formulations , reproductive disorders, 320t–322t

chemical marker compounds isolation, 314
,
314t

drugs and mechanism of action, 316t

herbs of, 310t

home, 309–310

lack of standardization, 310–319

modern system of medicine, 319–322

pharmacological activities, 322t–324t

plants distribution, 311f

red list category and criteria, 328–330

scientific basis of pairing, 315–319
,
315t

standardization steps, 314

substitutes, 313t
,
315–319
,
317t–318t

traditional system of medicine, 319f

Asparagus racemosus

adverse effect and precaution, 252

chemical constituents, 251–252

clinical studies, 252

cultivation, 247–248

habit and habitat, 247

macroscopic characterization, 247

microscopic characterization, 247

pharmacological activities, 248–250

traditional uses, 248

Asthma treatment

fenugreek, 143

Moringa oleifera, 82

Ayurvedic literature, 4
B
Bacognize, 120
BacoMind, 120–123
Bacopa monnieri

aglycone, 112–113

ayurvedic prescriptions, 111

bacosides, 112

bioactive phytochemicals, 111

biological activities, 118

chemical constituents, 112–113

bacopasides, 112–113

bacosaponins, 112–113

bacosides, 112
,
115

extraction methods, 113

preliminary phytochemical analysis, 113

commercial food products, 124

herbal formulations

Bacognize, 120

BacoMind, 120–123

CDRI-08 extract, 119–120
,
121t–122t
,
123–124

cognitive enhancers, 120

neuropharmacological effects, 111

neuroprotection, 118–119

plant morphology, 112

qualitative and quantitative analysis, 113–115

saponins

infrared (IR) spectroscopy, 116

NMR spectroscopy, 116–117

Bacopasaponin C, 117

Bacosides

bacoside A and B, 112–113

biological activities, 118

chemical constituents, 112

herbal formulations, 119–124

memory-enhancing effect, 118–119

neuroprotection, 118–119

quantitative determination, 113–115

Bioactive compounds, 4

Bacopa monnieri, 111

fenugreek, 135t

galangal, 96f

Moringa oleifera, 69–70

Myristica fragrans, 287–288

Piper nigrum (black pepper), 43

Rosmarinus officinalis, 150f
,
150t

turmeric, 15–16

Black pepper, See
Piper nigrum (black pepper)

Boswellia serrata gum resin extract

antiinflammatory properties, 31

benefits, 32

composition, 31

location, 31

as nutraceutical/functional food

anticancer properties, 36

antimicrobial properties, 37

antioxidant properties, 37

diabetes treatment, 36–37

gastrointestinal diseases, 35–36

inflammatory diseases, 34–35

neurological diseases, 32–34

turpentine source, 31

Boswellic acids, 31

Bovine coronavirus (BCV), 224

Brain-derived neurotrophic factor (BDNF), 32–33

Brain function and effect, ursolic acid (UA), 163

Bread, 357

Breast cancer treatment

galangal, 100

pomegranate, 181–182

Brief visuospatial memory test-revised (BVMT-R), 196–197

C

Camellia sinensis (tea)

antibacterial activity, 223

anticancer activity, 222–223

antihypertensive and cardiovascular activity, 225

antimicrobial activity, 224–225

antioxidant activity, 221–222

antiviral activity, 223–224

chemical constituents, 219

neuroprotective effects, 225

pharmacokinetics, 221

pharmacological activities, 221–225

phytochemicals, 219–221

weight management, 224

cAMP-response element-binding protein (CREB), 32–33

Cancer treatment, 348–349

Boswellia serrata gum resin extract, 36

curcumin/turmeric extract supplementation, 21–22

Cardioprotective activity

galangal, 101–102

pomegranate, 190

Cardiovascular disease (CVD), 348

Camellia sinensis (tea), 225

cinnamon, 267

ursolic acid (UA), 163

Carnosic acid

carotenoids like paprika stabilization, 158–159

cognition, 161–162

meat and meat products

cooked and smoked pork sausages, 158

cured cooked chicken sausage, 158

nitrite replacement, 157

nuggets, 158

oxidative reactions, 156

raw beef patties, 157

raw pork burger preservation, 157

sausages, 157

synthetic antioxidants, 156

vacuum-packed pork patties, 157

oil stability and deep frying process, 152–153

Center for Science in the Public Interest (CSPI), 373–374

Cereal-based foods, 18–19

Cereals, 357

Chemotherapy-induced peripheral neuropathy (CINP), Boswellia serrata gum resin extract, 33–34

Chinese food safety law, 340

Chronic variable stress (CVS) paradigm, Ocimum sanctum, 63

Chyawanprash, 4

Cinnamaldehyde, 260–261
,
264

Cinnamon

advanced glycation end products, 268

Alzheimer’s disease, 267

antibacterial activity, 265–266

anticancer activity, 266–267

antidiabetic activity, 263–264

antiinflammatory activity, 265

antioxidant activity, 262–263

antiulcer and gastric effect, 267

cardiovascular disease, 267

ceylon zeylanicum and cinnamon cassia, 259f

composition, 259–260

culinary uses, 268–269

leaves and barks, 257–258

natural compounds, 260–267

Parkinson’s disease, 267

species, 258–259

therapeutic uses, 260–267

Cinnamon cassia, 259

Cinnamon zeylanicum

advanced glycation end (AGE) products, 268

antidiabetic activity, 264

antioxidant activity, 262

chemical compounds, 260t

compositions, 259–260

Cognition

carnosic acid and rosmarinic acid, 161–162

Ocimum sanctum, 62–63

Colorectal cancer, curcumin/turmeric extract supplementation, 21–22

Compliant marketing, 377

Curcuma longa
, See
Turmeric

Curcumin

anticancer activities, 21–22

bioavailability, 16–17

cereal-based foods, 18–19

in dairy-based product, 19

inflammatory disorder treatment, 20–21

metabolic disorder control, 22

pharmacological indications, 23

vitiligo treatment, 23

Curcuminoids, 16f

bioavailability, 16–17

cancer treatment, 21–22

formulations, 20–23

functional foods

cereal-based foods, 18–19

dairy-based product, 19

phytochemicals, 19

inflammatory disorder treatment, 20–21

metabolic disorder treatment, 22

molecular targets, 17–18
,
18f

neurodegenerative disorder treatment, 22

solvent extraction, 16

D
Dairy-based product, curcumin, 19
Dammarane triterpenoid glycosides, 112
Dejerine–Sottas disease, curcumin, 22
Dementia, Ocimum sanctum, 62–63
Deoxyribonucleic acid (DNA) barcoding

Garcinia cambogia, 214–215

Depression, Piper nigrum (black pepper), 51–52

Diabetes mellitus (DM)

See also
Antidiabetic activity

Boswellia serrata gum resin extract, 36–37

curcumin supplementation, 22

Ocimum sanctum, 64–65

Dietary fibers, 8
,
136

Dietary Supplement Health and Education Act (DSHEA) 1994, 338–339
,
373–374

Dietary supplements, 6–8
,
7t

Dietary Supplements Regulations of 1985, 339–340

Diosgenin, 135
Diphenyl alkanes, 286f
Dyslipidemia, galangal, 101–102
E
Electrical resistance, 355–356
Ellagic acid, 173–174
Epicatechin (EC)

antimicrobial activity, 224–225

antioxidant activity, 222

Epicatechin gallate (ECG)

antibacterial activity, 223

antioxidant activity, 222

Epigallocatechin (EGC), 222

Epigallocatechin gallate (EGCG)

antibacterial activity, 223

anticancer activity, 222–223

antimicrobial activity, 224–225

antioxidant activity, 222

antiviral activity, 224

weight management, 224

Essential oil

Piper nigrum (black pepper), 52–53

Rosemary oil, See
Rosemary oil

European Food Safety Authority (EFSA), 341–342

Exercise activity and effect, ursolic acid (UA), 163

F
Fatty acids and esters, fenugreek, 136
Fenugreek

anthelmintic activity, 141–142

antibacterial and antifungal activity, 140

anticancer property, 137–138

antidiabetic activity, 138

antiobesity activity, 139

antioxidant property, 141

antiulcer property, 142

asthma treatment, 143

bioactive compounds, 135t

chemical constituents, 135t

functional food, 143–144

immunological activity, 139

lactation aid, 138–139

liver’s hepatotoxicity and genotoxicity, 141

morphology, 134

rheumatoid arthritis, 142–143

seeds and leaves, 133

yield, 133

Fermented foods, 356

Ferula asafoetida

See also
Asafoetida

main constituents, 234t

main fractions, 234t

Flavonoids, fenugreek, 136
Food Act of 2014, 339–340
Food herbs, 4–6
Food Hygiene Law, 340
Food preparation, 353
Food Safety and Standard Act (FSSA), 343

Food Safety and Standard Authority of India (FSSAI), 343

Food Safety Law, 340

Foods for specified health uses (FoSHU), 340–341

Food sterilization, 354

Food with Nutrient Function Claims (FNFC), 340–341

Fritillaria roylei, 328
Furostanol, 135
G

Galactagogue effects, Asparagus racemosus, 249

Galactomannan, 136

Galangal

anticancer activity, 100–101

antiinflammatory agent, 101

antimicrobial activity, 103–104

antioxidant potential, 102

antiviral effect, 103

bioactive components, 96f

biological activities, 100–104

cardioprotective agent, 101–102

formulations, 98–100

functional foods

flavorings, 98–99

stimulant and carminative bioactivities, 99

therapeutic effects, 98–99

immune stimulant action, 104

liver health, 102

neuroprotective and memory enhancing activity, 103

packaging material, 99–100

phenolic substances, 95

phytochemicals, 97

phytochemistry, 95–97

reproductive agent, 104

rhizome, 96–97

traditional medicine, 95–96

Gamma linolenic acid (GLA), 350

Garcinia cambogia, 212f

adulterants, 212f

antiadiposity properties, 214–215

hydroxycitric acid (HCA), 211–213

mechanism of action, 213–214

organic acids, 212f

pharmacological activities, 213–214

phytochemical ingredients, 211–212

Gastrointestinal health

asafoetida, 236

Boswellia serrata gum resin extract, 35–36

cinnamon, 267

Piper nigrum (black pepper), 51

Genotoxicity, fenugreek, 141

Global nutraceutical industry

high health-care cost, 351

high personnel income, 351–352

lifestyles and sedentary work environments, 351–352

noncommunicable diseases

antiangiogenic medications, 350

cancer, 348–349

cardiovascular disease (CVD), 348

diabetes, 349

drug toxicology, 350–351

gamma linolenic acid (GLA), 350

obesity, 349–350

probiotics, 349

viral infections, 349

technologies, 352–358
,
355t

Global nutraceutical market, 2
,
2f

Glutamate-induced neuronal cell damage, asafoetida, 235

Green pepper, 41

Green tea

antibacterial activity, 223

anticancer activity, 222–223

antihypertensive and cardiovascular activity, 225

antimicrobial activity, 224–225

antioxidant activity, 221–222

antiviral activity, 223–224

pharmacokinetics, 221

weight management, 224

Gut microbiome research, 357
H
Habenaria edgeworthii (Riddhi), 329–330
Health Claim, 9
Hepatoprotective activity

asafoetida, 237

Moringa oleifera, 83

Myristica fragrans, 296

pomegranate, 190

Hepatotoxicity, fenugreek, 141
Herbal medications, 358
Herbals, 4–6

Holy basil, See
Ocimum sanctum

Hydroxycitric acid (HCA), 212–213

Hydroxycitric acid Super CitriMax (HCA-SX), 212–213

I

Immune stimulant action, galangal, 104

Immunoadjuvant effects, Asparagus racemosus, 250

Immunological activity, fenugreek, 139

Immunomodulatory activity

Asparagus racemosus, 250

ursolic acid (UA), 162

Inflammatory diseases

Boswellia serrata gum resin extract, 34–35

curcuminoids, 20–21

Insoluble dietary fibers, 8
Isoflavones, 363–364
J
Jujubogenin, 112f
K
Kamya, 4
L
Lactation aid, fenugreek, 138–139

Lignans, 283
,
285f

Lipid-lowering effect, pomegranate, 185
Lipid metabolism illnesses, galangal, 101–102
Lipids, fenugreek, 136
Lithiatic activity, Asparagus racemosus, 250
Liver health, galangal, 102
Lupus nephritis, curcuminoids, 23
M
Mahameda, 329

Marketed nutraceuticals, 2
,
2f

Medhya rasayana, 111
Medicinal herbs, 4–6
Medicinal plants, 5t–6t
Memory-enhancing activity, asafoetida, 235
Metabolic activity, asafoetida, 239

Metabolic disorders, curcumin supplementation, 22

Metchnikoff’s longevity theory, 356–357
Microangiopathy, curcumin supplementation, 22
Microstylis muscifera, 329
Milk products, 356
Minerals

fenugreek, 136

resources, plants, 7t

Molecular biology technologies, 375–376

Moringa oleifera

amino acid composition, 72t

antiasthmatic properties, 82

anticancer properties, 80–81

antidiabetic properties, 81–82

antiinflammatory properties, 77–78

antimicrobial properties, 75–77

antioxidant properties, 78–79

bioactive compounds, 69–70

composition, 70–72
,
71t

fatty acid composition, 73t

hepatoprotective properties, 83

mineral composition, 72
,
74t

neuroprotective properties, 83–84

nutritional value, 70–72

phytochemicals, 69–70

species, 69

traditional food and uses, 76t

fresh leaves, 75

plant, 75

seeds, 75

tree parts, 69–70

Myristica fragrans

applications, 288–300

bioactive components, 287–288

composition, 281
,
282t

culinary and HORECA products, 290–291

fruit

morphology and pharmacognostical details, 280–281
,
280f

phytochemistry, 281–286
,
284t

rind, 280–281
,
282t

morphology, 280–281

nutrition source, 289–290

pharmacognostical details, 280–281

taxonomy, 280

therapeutic benefits, 291–300
,
292t–295t

toxicity aspects, 301–302

N
Naimittika, 4
Nanotechnology, Punica granatum, 192

Natural Health Products Regulations (NHPR), 339

Nectandrin B, 297

antiageing, 300

antiobesity, 300

hepatocyte-protective effect, 297

therapeutic activities, 292t–295t

Neolignans, 283
,
285f

Nephroprotective activity, pomegranate, 190

Neurodegenerative disorders, curcuminoids, 22

Neurological diseases, Boswellia serrata gum resin extract, 32–34

Neuromodulatory potential, Ocimum sanctum, 64

Neuroprotective effects

asafoetida, 235–236

Asparagus racemosus, 250

Bacopa monnieri, 118–119

Moringa oleifera, 83–84

pomegranate, 182–183

New Dietary Ingredient Notification process, 338–339

Nonthermal technologies, 352
,
357

Nutmeg

antiageing, 300

antibacterial activity, 290
,
298–299

anticancer activity, 299

antidiabetic activity, 297

antiinflammatory activity, 298

antiobesity, 300

composition, 281
,
282t

hepatoprotective effect, 296

morphology, 281

nutrient value, 289t
,
290

nutrition source, 289

oleoresin, 290–291

phytoconstituents, 284t

secondary metabolites, 287

source, 289

terpenes, 283–286

toxicity aspects, 301–302

traditional uses, 291–292
,
292t–295t

Nutraceuticals, definition, 1
Nutraceuticals industries and challenges, 10
Nutrigenomics, 375–376
O
Obesity, 349–350

See also
Antiobesity activity

Ocimum sanctum

antifertility potential, 65

beneficial effects, 62–66

chemical composition, 61

composite extracts of leaves (CLEs), 64–65

essential oil extract, 62

flowers and fruits, 61

hormonal effect, 65

hormones, 65

for lead acetate-induced hepatotoxicity and nephrotoxicity, 65–66

medicinal properties, 62

neurological treatment, 62–63

phytochemicals, 61–62

polyphenols, 62

secondary metabolites, 62

Ohmic heating, 355–356

Osteoarthritis (OA), curcumin formulations, 20–21

P
Paprika, 158–159
Parkinson’s disease, cinnamon, 267
Pepper

black, See
Piper nigrum (black pepper)

classification, 41

green pepper, 41

white, 41

Peripheral neuropathy, asafoetida, 235
Phala Ghrita, 4
Pharmacological activities

Ashtawarga plants, 322t–324t

Asparagus racemosus, 248–250

Camellia sinensis (tea), 221–225

Garcinia cambogia, 213–214

pomegranate, 181–191

Phenolic compounds, Myristica fragrans fruit, 283

Phenylpropanoids, 283
,
285f

Phytochemicals

Camellia sinensis (tea), 219–221

galangal, 97

Garcinia cambogia, 211–212

Moringa oleifera, 69–70

pomegranate, 174–181

Phytoestrogenic effects, Asparagus racemosus, 249

Piperine, 41

anticancer property, 46–47

antidepressant activity, 51–52

antiinflammatory property, 47

antimicrobial property, 47–51

antiobesity property, 51

antioxidant property, 45–46

biological activity, 45f

chemical structures and biological activities, 48t–50t

environmental factors, 41–42

gastrointestinal health, 51

therapeutic properties, 41–42

Piper nigrum (black pepper)

bioactive components, 43

culinary and ethnomedical uses, 53–54

essential oil, 52–53

fruit, 42

gastrointestinal activity, 41

gastrointestinal health, 51

human diet, 41

nutritional and phytonutrient composition, 42–43

and piper species

anticancer property, 46–47

antiinflammatory property, 47

antimicrobial property, 47–51

antiobesity property, 51

antioxidant property, 45–46

anxiolytic property, 51–52

quality specifications, 43

recommended dosage, 43
,
44t

taxonomy, 42

Poisonous herbs, 4–6

Pomegranate

antiacnes activity, 189

antibacterial activity, 185

anticancer activity, 181–182

anticoccidial activity, 186

antidiabetic activity, 188–189

antiinflammatory activity, 186–188

antileishmanial activity, 186

antilice and antidandruff activity, 189

antioxidant activity, 183–185

antiviral activity, 185–186

cardioprotective activity, 190

clinical trials, 196–197

cultivation, 171–172

economic importance, 174

fruits, 171–172

health benefits, 173f

hepatoprotective activity, 190

industrial uses, 192–193

lipid-lowering effect, 185

nephroprotective activity, 190

neuroprotective effect, 182–183

nutritional and medicinal importance, 173–174

pharmacological activities, 181–191

phytochemicals, 174–181
,
175t–176t

bark extract, 179–181

environment pollutant management, 191–192

leaf ethanolic extract, 174

peel extract, 177

pomegranate flower, 179

pomegranate juice, 177

seed residue, 178–179

pregnancy complication treatment, 189

toxicity, 197

tree, 171–172

vasculoprotective effect, 190–191

Post-biotics, 357

Prebiotics, 8

Pregnancy complication treatment, pomegranate, 189

Probiotics, 8
,
349–350

Product strategy, 378

Prophylactic and protective effect, Myristica fragrans fruit, 296–297

Proteins, 9
Pseudojujubogenin, 112f
Psoriasis, turmeric, 23
Punica granatum

cultivars, 171

morphology, 172

nanotechnology, 192

patents granted, 193

production, 174

R
Radiofrequency drying, 355
Rasayana, 4
Regulatory aspects, 9

in Association of South East Asian Nation (ASEAN) countries, 342

in Australia and New Zealand, 339–340

in Canada, 339

in China, 340

in European Union, 341–342

in India, 343

in Japan, 340–341

need for, 338

in United States of America, 338–339

Relaxant effects, asafoetida, 236–237

Reproductive effects

asafoetida, 239

galangal, 104

Rheumatoid arthritis, fenugreek, 142–143
Roscoea purpurea, 328
Rosemary oil

aromatherapy and cosmetics, 150–152

carnosic acid

carotenoids like paprika stabilization, 158–159

cognition, 161–162

meat and meat products, 156–158

oil stability and deep frying, 152–153

food additives specifications, 151t

gas chromatography profile, 152t

manufacturing regulations E392, 151t

natural antioxidants, 149–150

oil stability

antioxidant effects, 153

bioactives, 153

meat-based food products, 155t–156t

multicomponent antioxidant formulation, 153–156

rosmarinic acid

beverage drinks, 159–161

cognition, 161–162

specification, 150t

ursolic acid, 162–163

Rosmarinic acid

beverage drinks

color stability, 160–161

shelf life extension, 159–160

cognition, 161–162

specification, 150t

Rosmarinus officinalis, 150f
,
150t

See also
Rosemary oil

S

Saponins

Bacopa monnieri, 116–117

fenugreek, 135

Selective reminding test (SRT), 196–197
Skin disorders, turmeric, 23
Soluble dietary fibers, 8
Spirostanol, 135
Steroidal compounds, fenugreek, 135

Stroke, Boswellia serrata gum resin extract, 32

Supercritical carbon dioxide extraction, 354

Supercritical fluid extraction (SCFE), curcuminoid, 16

Supply chain, 378
T
Tambuli, 54
Terpenes, Myristica fragrans fruit, 283–286
Therapeutic Goods Act (TGA), 339

Total neuropathy score clinical version (TNSc), 33–34

Toxicity aspects

Myristica fragrans, 301–302

nutmeg, 301–302

Traditional nutraceuticals and products

accessibility, 372–373

affluence, 371–372

affordability, 372

awareness, 372

challenges, 373

China, 375

compliant marketing, 377

condition-specific needs, 372

deal making, 378

enhancement needs, 373

European Union (EU), 374

foundation needs, 372

functional foods

health benefits, 364–367
,
366t

tomato, 364

types, 366t

global market, 369–370

global regulations, 374–375

health benefits, 369

India, 375

Japan, 375

motivators, 370–371

nutraceuticals, 363f

vs. functional foods, 367
,
368f
,
368t

herbals with health benefits, 365t

isoflavones, 363–364

product strategy, 378

regulation, 377

regulatory and perception issues, 373–374

supply chain, 378

technology, 377

toxicity potential, 375

United States, 374

Trigonella foenum graecum, 134f

See also
Fenugreek

Trigonelline, 135–136

Trikatu, 53–54

Tulsi, See
Ocimum sanctum

Turmeric

applications, 15

bioactive components, 15–16

chemical composition, 15–16

curcumin, See
Curcumin

curcuminoids, See
Curcuminoids

isolation, 15–16

phytochemistry, 15–16

traditional systems, 15

U

Ulcerative colitis, curcumin/turmeric extract supplementation, 21

Ultrasound, 354

Ursolic acid (UA)

anticancerous activity, 162

antimicrobial effect, 162

antiobesity/antidiabetic activity, 163

brain function and effect, 163

cardiovascular disease and effect, 163

exercise activity and effect, 163

immunity-enhancing effect, 162

V

Vasculoprotective effect, pomegranate, 190–191

Vitamins, fenugreek, 136
W
Wechsler Memory Scale-Revised (WMS-R), 33

Weight management, Camellia sinensis (tea), 224

White pepper, 41