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Edited by
Javed Ahmad
College of Pharmacy,
Najran University,
KSA
Co-editor
Javed Ahamad
Department of Pharmacognosy,
Faculty of Pharmacy,
Tishk International University,
Kurdistan Region, Erbil,
Iraq
Bioactive Phytochemicals:
Drug Discovery to Product
Development
%LRDFWLYH3K\WRFKHPLFDOV'UXJ'LVFRYHU\WR3URGXFW'HYHORSPHQW
Co-editor: Javed Ahamad
ISBN (Online): 978-981-14-6448-5
ISBN (Print): 978-981-14-6446-1
© 2020, Bentham eBooks imprint.
Editor: Javed Ahmad
ISBN (Print): 978-981-14-6447-8
Published by Bentham Science Publishers Pte. Ltd. Singapore. All Rights Reserved.
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CONTENTS
FOREWORD ........................................................................................................................................... i
PREFACE ................................................................................................................................................ ii
LIST OF CONTRIBUTORS .................................................................................................................. iii
CHAPTER 1 DRUG DISCOVERY FROM PLANT SOURCES: SCOPE, APPROACH AND
CHALLENGES ....................................................................................................................................... 1
Javed Ahmad and Javed Ahamad
1. INTRODUCTION ...................................................................................................................... 1
2. ADVANTAGES OF DRUG DISCOVERY FROM NATURAL RESOURCES ................... 3
3. ROLE OF MEDICINAL PLANTS IN DRUG DISCOVERY ................................................ 4
4. STEPS OF DRUG DISCOVERY FROM HIGHER PLANTS .............................................. 7
5. DRUG DISCOVERY FROM HIGHER PLANTS: APPROACHES .................................... 7
5.1. Ethno-Pharmacological Approach ................................................................................... 8
5.2. Follow-up of Pharmacological Reports ........................................................................... 9
5.3. Random Selection Followed by Phytochemical Screening ............................................. 9
5.4. Random Selection Followed by Pharmacological Screening .......................................... 10
6. CHALLENGES IN DRUG DISCOVERY FROM HIGHER PLANTS ................................ 10
CONCLUDING REMARKS ......................................................................................................... 11
ABBREVIATIONS ......................................................................................................................... 11
CONSENT FOR PUBLICATION ................................................................................................ 11
CONFLICT OF INTEREST ......................................................................................................... 12
ACKNOWLEDGEMENTS ........................................................................................................... 12
REFERENCES ............................................................................................................................... 12
CHAPTER 2 CULTIVATION, COLLECTION AND PROCESSING OF MEDICINAL
PLANTS ................................................................................................................................................... 14
Omji Porwal, Sachin Kumar Singh, Dinesh Kumar Patel, Saurabh Gupta, Rahul
Tripathi and Shankar Katekhaye
1. IMPORTANCE OF MEDICINAL PLANTS ......................................................................... 15
2. CULTIVATION OF MEDICINAL PLANTS .......................................................................... 15
2.1. Method of Propagation .................................................................................................... 16
2.1.1. Sexual or Seed Propagation ............................................................................... 16
2.1.2. Asexual or Vegetative Propagation ................................................................... 16
3. FACTORS AFFECTING CULTIVATION ............................................................................. 16
3.1. Soil ................................................................................................................................... 17
3.2. Altitude, Temperature and Humidity ............................................................................... 17
3.3. Rainfall and Irrigation ...................................................................................................... 18
3.4. Fertilizers ......................................................................................................................... 18
3.5. Pest and Pest Management ............................................................................................... 19
3.5.1. Methods of Pest Control ...................................................................................... 19
3.5.1.1. Natural Controls .............................................................................................. 19
3.5.1.2. Artificial Control .............................................................................................. 20
3.5.1.3. Machine-like Mechanical Control ................................................................... 20
3.5.1.4. Agricultural control ......................................................................................... 20
3.5.1.5. Chemical Controls ........................................................................................... 20
4. ROLE OF PLANT GROWTH REGULATORS ..................................................................... 21
4.1. Auxins .............................................................................................................................. 21
4.2. Gibberellins ...................................................................................................................... 21
4.3. Cytokinins ........................................................................................................................ 22
4.4. Abscisic Acid ................................................................................................................... 22
4.5. Ethylene ........................................................................................................................... 22
4.6. Other Growth Regulators ................................................................................................. 23
5. ORGANIC FARMING .............................................................................................................. 23
6. COLLECTION OF MEDICINAL PLANTS ........................................................................... 24
7. HARVESTING OF MEDICINAL PLANTS ........................................................................... 24
8. DRYING OF MEDICINAL PLANTS ...................................................................................... 25
8.1. Natural Drying ................................................................................................................. 25
8.2. Artificial Drying ............................................................................................................... 26
8.2.1. Tray Drying ......................................................................................................... 26
8.2.2. Vacuum Drying ................................................................................................... 26
8.2.3. Spray Drying ....................................................................................................... 26
9. GARBLING OF MEDICINAL PRODUCTS .......................................................................... 26
10. PACKING OF MEDICINAL PRODUCTS ........................................................................... 26
11. STORAGE AND PRESERVATION OF MEDICINAL PLANTS ...................................... 27
CONCLUDING REMARKS ......................................................................................................... 28
ABBREVIATIONS ......................................................................................................................... 28
CONSENT FOR PUBLICATION ................................................................................................ 29
CONFLICT OF INTEREST ......................................................................................................... 29
ACKNOWLEDGEMENTS ........................................................................................................... 29
REFERENCES ............................................................................................................................... 29
CHAPTER 3 EXTRACTION OF BIOACTIVE PHYTOCHEMICALS ........................................ 31
Javed Ahamad, Naila Hassan Ali Alkefai and Shehla Nasar Mir Najibullah
1. INTRODUCTION ...................................................................................................................... 31
2. STEPS INVOLVED IN THE EXTRACTION OF MEDICINAL PLANTS ........................ 33
2.1. Drying and Grinding of Plant Materials .......................................................................... 33
2.2. Selection of Suitable Solvents ......................................................................................... 33
2.3. Extraction Process ............................................................................................................ 33
2.4. Filtration ........................................................................................................................... 34
2.5. Concentration ................................................................................................................... 34
2.6. Drying of Extract ............................................................................................................. 34
3. FACTORS AFFECTING EXTRACTION PROCESS ........................................................... 34
4. METHODS OF EXTRACTION ............................................................................................... 35
4.1. Classical Methods of Extraction ...................................................................................... 35
4.1.1. Maceration ......................................................................................................... 35
4.1.2. Decoction ........................................................................................................... 36
4.1.3. Percolation ......................................................................................................... 36
4.1.4. Reflux Extraction ................................................................................................ 36
4.1.5. Soxhlet Extraction .............................................................................................. 37
4.1.6. Isolation of Essential Oils .................................................................................. 37
4.2. Modern Methods of Extraction ........................................................................................ 38
4.2.1. Ultrasound-Assisted Extraction (UAE) ............................................................... 39
4.2.2. Microwave-Assisted Extraction (MAE) .............................................................. 39
4.2.3. Pressurized Liquid Extraction (PLE) .................................................................. 40
4.2.4. Supercritical Fluid Extraction (SFE) ................................................................. 40
4.2.5. Enzyme Assisted Extraction (EAE) ..................................................................... 40
5. CASE STUDIES: BIOACTIVE PHYTOCHEMICAL ISOLATION AND
PURIFICATION ............................................................................................................................. 42
5.1. Piperine ............................................................................................................................ 42
5.2. Piperine Isolation ............................................................................................................. 42
5.3. Quinine ............................................................................................................................. 43
Isolation ......................................................................................................................... 43
5.4. Solasodine ........................................................................................................................ 43
Isolation ......................................................................................................................... 44
5.5. Caffeine ............................................................................................................................ 44
Isolation ......................................................................................................................... 44
5.6. Starch ............................................................................................................................... 45
5.7. Menthol ............................................................................................................................ 46
Isolation ......................................................................................................................... 46
5.8. Artemisinin ...................................................................................................................... 46
Isolation ......................................................................................................................... 47
5.9. Reserpine .......................................................................................................................... 47
Isolation ......................................................................................................................... 47
5.10. Digitoxin ........................................................................................................................ 48
Isolation ......................................................................................................................... 48
6. CONCLUDING REMARKS ..................................................................................................... 49
CONSENT FOR PUBLICATION ................................................................................................ 49
CONFLICT OF INTEREST ......................................................................................................... 49
ACKNOWLEDGEMENTS ........................................................................................................... 49
REFERENCES ............................................................................................................................... 49
CHAPTER 4 ISOLATION AND PURIFICATION OF BIOACTIVE PHYTOCHEMICALS .... 54
1. INTRODUCTION ...................................................................................................................... 55
2. TECHNIQUES OF NATURAL PRODUCT ISOLATION .................................................... 56
2.1. Counter-Current Chromatography (CCC) ....................................................................... 56
2.2. Column Chromatography (CC) ........................................................................................ 59
2.3. Medium Pressure Liquid Chromatography (MPLC) ....................................................... 62
2.4. Preparative HPLC ............................................................................................................ 64
2.5. Preparative Gas Chromatography (Prep-GC) .................................................................. 65
2.6. Preparative TLC (Prep-TLC) ........................................................................................... 65
CONCLUDING REMARKS ......................................................................................................... 66
ABBREVIATIONS ......................................................................................................................... 66
CONSENT FOR PUBLICATION ................................................................................................ 66
CONFLICT OF INTEREST ......................................................................................................... 66
ACKNOWLEDGEMENTS ........................................................................................................... 66
REFERENCES ............................................................................................................................... 67
CHAPTER 5 SPECTROSCOPIC TECHNIQUES FOR THE STRUCTURAL
CHARACTERIZATION OF BIOACTIVE PHYTOCHEMICALS .................................................. 73
Showkat R. Mir, Tara Fuad Tahir, Javed Ahamad, Raad A Kaskoos, Naila Hassan
Ali Alkefai and Abdul Samad
1. INTRODUCTION ...................................................................................................................... 74
2. STRUCTURAL ELUCIDATION OF PHYTOCONSTITUENTS ........................................ 74
2.1. UV-Visible Spectroscopy ................................................................................................ 75
2.2. Fourier Transform Infrared (FT-IR) Spectroscopy .......................................................... 80
2.3. NMR Spectroscopy .......................................................................................................... 82
2.4. Mass Spectrometry ........................................................................................................... 86
ESI MS Mass Spectrometry .................................................................................................... 87
3. CASE STUDIES: SOME EXAMPLES OF STRUCTURAL CHARACTERIZATION OF
NATURAL PRODUCTS ................................................................................................................ 88
Kamran Javed Naquvi, Javed Ahamad, Raad A Kaskoos, Naila Hassan Ali Alkefai, Afrin Salma and
Showkat R. Mir
3.1. Erythrocentaurine (EC) .................................................................................................... 88
3.2. Octatriacontanoic Acid (OA) ........................................................................................... 89
3.3. Norgadosic Acid (NA) ................................................................................................... 91
3.4. Isoalantolactone (IL) ........................................................................................................ 92
3.5. Quercetin (QC) ................................................................................................................. 94
CONCLUDING REMARKS ......................................................................................................... 95
ABBREVIATIONS ......................................................................................................................... 96
CONSENT FOR PUBLICATION ................................................................................................ 96
CONFLICT OF INTEREST ......................................................................................................... 96
ACKNOWLEDGEMENTS ........................................................................................................... 96
REFERENCES .............................................................................................................................. 97
CHAPTER 6 PHARMACOLOGICAL EVALUATION OF HERBAL MEDICINE .................... 100
Subasini Uthirapathy, Javed Ahamad, Jaswanth Albert and Govind Prasad Dubey 100
1. INTRODUCTION ...................................................................................................................... 101
2. COMMON LABORATORY ANIMALS ................................................................................. 102
2.1. Albino Rats (Rattus Norvegicus) ..................................................................................... 102
2.2. Albino Mice (Mus musculus) .......................................................................................... 103
2.3. C57BL/6 (B6) Black Mice ............................................................................................... 103
2.4. Guinea Pigs (Cavia porcellus) ......................................................................................... 103
2.5. Rabbits (Lupas cuniculus) ................................................................................................ 103
3. ETHICS OF ANIMAL EXPERIMENTATION ...................................................................... 103
3.1. Objectives of Anesthesia .................................................................................................. 103
3.2. Pre-Anesthetic Medications ............................................................................................. 104
3.3. Drugs ................................................................................................................................ 104
3.4. Standard Bleeding Techniques ........................................................................................ 104
3.5. Euthanasia ........................................................................................................................ 106
3.6. Types of Euthanasia ......................................................................................................... 106
3.6.1. Physical Methods ............................................................................................... 106
3. PHARMACOLOGICAL EVALUATION METHODS .......................................................... 107
3.1. Evaluation of Analgesic Activity of Herbal Medicine .................................................... 107
3.1.1. Hotplate Analgesiometer Method ...................................................................... 107
3.1.2. Tail Clip Method ................................................................................................ 108
3.1.3. Tail Flick Method ............................................................................................... 108
3.1.4. Acetic Acid-Induced Writhing ............................................................................ 108
3.2. Evaluation of Anti-Inflammatory Activity of Herbal Medicine ...................................... 109
3.3. Evaluation of Pupil Size Effects of Herbal Medicine ...................................................... 110
3.4. Evaluation of the Hypnotic Effect of Herbal Medicine ................................................... 110
3.5. Evaluation of Muscle Relaxant Activity of Herbal Medicine ......................................... 111
3.6. Evaluation Of Anti-Anxiety Effect Of Herbal Medicine ................................................. 111
3.7. Evaluation of Local Anesthetic Activity of Herbal Medicine ......................................... 111
3.8. Evaluation of Anti-Peptic-Ulcer Activity of Herbal Medicine ........................................ 112
3.8.1. Ethanol-Induced Ulcer Model ............................................................................ 112
3.8.2. Pylorus Ligation Method (Shay Method) ........................................................... 112
3.8.3. Indomethacin Induced Ulcer .............................................................................. 112
3.8.4. Assessment of Gastric Mucosal Lesions ............................................................ 113
3.9. Evaluation of Anti-Depressant Effect of Herbal Medicine .............................................. 113
3.10. Evaluation of Anti-AtherosclerosisActivity of Herbal Medicine .................................. 114
3.11. Evaluation of Anti-diabetic Activity of Herbal Medicine ............................................. 115
3.12. Evaluation ofAnti-asthmatic Effect of Herbal Medicine ............................................... 115
3.13. Evaluation of Locomotor Activity of Herbal Medicine ................................................. 116
CONCLUSION ............................................................................................................................... 116
CONSENT FOR PUBLICATION ................................................................................................ 117
CONFLICT OF INTEREST ......................................................................................................... 117
ACKNOWLEDGEMENTS .......................................................................................................... 117
ABBREVIATIONS ......................................................................................................................... 117
REFERENCES .............................................................................................................................. 117
CHAPTER 7 PRODUCT DEVELOPMENT OF HERBAL MEDICINE ....................................... 121
Ahmed Nawaz Khan, Chandra Kala and Javed Ahmad
1. INTRODUCTION ...................................................................................................................... 121
1.1. Scope of Herbal Medicinal Products ............................................................................... 123
2. CHALLENGES IN HERBAL MEDICINAL PRODUCT DEVELOPMENT ..................... 123
3. DOSAGE FORMS OF HERBAL MEDICINAL PRODUCTS .............................................. 127
3.1. Solid Dosage Form .......................................................................................................... 128
3.2. Liquid Dosage Form ........................................................................................................ 129
3.3. Semi-Solid Dosage Forms ............................................................................................... 130
4. RECOMMENDATIONS IN HERBAL MEDICINAL PRODUCT DEVELOPMENT ...... 131
CONCLUSION AND FUTURE DIRECTIONS .......................................................................... 138
ABBREVIATIONS ......................................................................................................................... 139
CONSENT FOR PUBLICATION ................................................................................................ 140
CONFLICT OF INTEREST ......................................................................................................... 140
ACKNOWLEDGEMENTS ........................................................................................................... 140
REFERENCES ............................................................................................................................... 140
CHAPTER 8 QUALITY CONTROL OF HERBAL MEDICINAL PRODUCTS ......................... 143
Javed Ahamad, Esra T. Anwer, Muath Sh. Mohammed Ameen, Jamia Firdous and
Nehal Mohsin
1. INTRODUCTION ...................................................................................................................... 144
2. THE NEED OF STANDARDIZATION ................................................................................... 145
3. METHODS OF STANDARDIZATION OF HERBAL MEDICINAL PRODUCTS ........... 145
3.1. Botanical Parameters ....................................................................................................... 145
3.1.1. Morphological Characters .................................................................................. 145
3.1.2. Powder Microscopy ............................................................................................ 145
3.2. Physico-Chemical Parameters ......................................................................................... 146
3.2.1. Extractive Values ................................................................................................ 146
3.2.2. Ash Value ............................................................................................................ 146
3.2.3. Loss on Drying .................................................................................................... 147
3.2.4. Foreign Organic Matter ...................................................................................... 147
3.2.5. Swelling Index ..................................................................................................... 148
3.2.6. Foaming Index .................................................................................................... 148
3.2.7. Fat Content .......................................................................................................... 148
3.2.8. Resin Content ...................................................................................................... 148
3.2.9. Total Flavonoid Content ..................................................................................... 149
3.2.10. Total Phenolic Content ..................................................................................... 149
3.3. General Quality Parameters ............................................................................................. 150
3.3.1. Fluorescence Analysis ......................................................................................... 150
3.3.2. Powdered Drug Reaction with Chemical Reagents ............................................ 150
3.4. Phytochemical Screening ................................................................................................. 150
3.5. Determination of Toxic Residues .................................................................................... 151
3.5.1. Pesticide Residues ............................................................................................... 151
3.5.2. Aflatoxin Residue ................................................................................................ 151
3.5.3. Microbial Load .................................................................................................... 152
3.5.4. Heavy Metal Toxicity .......................................................................................... 152
3.6. Chromatographic Evaluation of Herbal Medicinal Products ........................................... 152
2.6.1. Thin Layer Chromatography and RF Values ...................................................... 152
3.6.2. High Performance Thin Layer Chromatography (HPTLC) ................................ 153
3.6.3. High Pressure Liquid Chromatography (HPLC) ................................................ 153
3.6.4. Gas Chromatography-Mass Spectrometry (GC-MS) ......................................... 153
3.6.5. Liquid Chromatography-Mass Spectrometry (LC-MS) ...................................... 154
CONCLUDING REMARKS ......................................................................................................... 154
ABBREVIATIONS ......................................................................................................................... 155
CONSENT FOR PUBLICATION ................................................................................................ 155
CONFLICT OF INTEREST ......................................................................................................... 155
ACKNOWLEDGEMENTS ........................................................................................................... 155
REFERENCES ............................................................................................................................... 155
CHAPTER 9 REGULATORY PERSPECTIVES OF HERBAL MEDICINAL PRODUCTS ..... 158
Faraat Ali, Shaik Khasimbi, Kamna Sharma, Manisha Trivedi, Asad Ali and Javed
Ahmad
1. INTRODUCTION ...................................................................................................................... 159
2. NEED AND CHALLENGES IN HERBAL PRODUCT REGULATIONS .......................... 161
3. INTERNATIONAL REGULATORY SCENARIO ................................................................ 163
3.1. The International Council for Harmonization .................................................................. 163
3.2. World Health Organization (WHO) ................................................................................. 164
3.3. United States Food and Drug Administration (USFDA) ................................................. 164
3.4. European Medicines Agency (EMA) ............................................................................... 165
4. REGULATORY GUIDELINES FOR HERBAL PRODUCTS ............................................. 165
4.1. Guidelines for Regulated Market ..................................................................................... 165
4.2. Guidelines for the Semi-regulated Market ....................................................................... 166
4.3. Guidelines for Non-regulatory Market ............................................................................ 166
5. RECOMMENDATIONS ON HERBAL MEDICINE IN PHARMACOPOEIAS ............... 166
5.1. United States Pharmacopoeia .......................................................................................... 166
5.2. Indian Pharmacopoeia ...................................................................................................... 167
5.3. Chinese Pharmacopoeia ................................................................................................... 170
5.4. European Union Pharmacopoeia ...................................................................................... 171
CONCLUSION ............................................................................................................................... 172
ABBREVIATIONS ......................................................................................................................... 172
CONSENT FOR PUBLICATION ................................................................................................ 172
CONFLICT OF INTEREST ......................................................................................................... 172
ACKNOWLEDGEMENTS ........................................................................................................... 173
REFERENCES ............................................................................................................................... 173
CHAPTER 10 PHYTOCHEMICALS FOR THE TREATMENT OF HUMAN DISEASES ........ 176
1. INTRODUCTION ...................................................................................................................... 177
2. HERBAL MEDICINES WITH ANTIDIABETIC ACTIVITY ............................................. 177
2.1. Current Therapies for Diabetes Mellitus .......................................................................... 179
2.2. Bioactive Phytochemicals with Antidiabetic Activity ..................................................... 180
2.2.1. Charantin ............................................................................................................ 180
2.2.2. Swertiamarin ....................................................................................................... 180
2.2.3. Gymnemic Acids .................................................................................................. 181
2.2.4. Oleuropein ........................................................................................................... 182
2.2.5. Berberine ............................................................................................................. 182
Javed Ahamad, Subasini Uthirapathy, Kamran Javed Naquvi, Muath Sh. Mohammed Ameen,
Esra T. Anwer, Abdul Samad and Mohammad Shabib Akhtar
2.2.6. 4-Hydroxyisoleucine ........................................................................................... 183
2.2.7. Epicatechin .......................................................................................................... 184
2.2.8. β-Sitosterol .......................................................................................................... 184
3. HERBAL MEDICINES WITH ANTICANCER ACTIVITY ................................................ 186
3.1. Bioactive Phytochemicals with Anticancer Activity ....................................................... 186
3.1.1. Vincristine and Vinblastine ................................................................................. 186
3.1.2. Podophyllotoxins ................................................................................................. 187
3.1.3. Taxanes ............................................................................................................... 188
3.1.4. Camptothecins ..................................................................................................... 188
3.1.5. Colchicine ........................................................................................................... 189
3.1.6. Resveratrol .......................................................................................................... 189
4. HERBAL MEDICINES WITH ANTI-INFLAMMATORY AND ANTIOXIDANT
POTENTIAL ................................................................................................................................... 191
4.1. Current Therapies for Inflammation ................................................................................ 192
4.1. Bioactive Phytochemicals with Anti-inflammatory and Antioxidant Activity ................ 192
5. HERBAL MEDICINES WITH CARDIOPROTECTIVE ACTIVITY ................................ 194
5.1. Current Therapies for Treatment of Cardiovascular Diseases ......................................... 196
5.1. Medicinal Plants and Bioactive Phytochemicals for Treatment of Cardiovascular
Diseases ................................................................................................................................... 196
6. HERBAL MEDICINES WITH ANTIOBESITY ACTIVITY ............................................... 198
6.1. Current Strategies for Management of Obesity ............................................................... 198
6.2. Role of Herbal Medicines for the Treatment of Obesity ................................................. 199
CONCLUDING REMARKS ......................................................................................................... 201
ABBREVIATIONS ......................................................................................................................... 201
CONSENT FOR PUBLICATION ................................................................................................ 202
CONFLICT OF INTEREST ......................................................................................................... 202
ACKNOWLEDGEMENTS ........................................................................................................... 202
REFERENCES ............................................................................................................................... 202
SUBJECT INDEX
.................................................................................................................................... 214
i
FOREWORD
Plants have always considered been as ‘Nature’s Library’ for new chemical entities and one
of the major resources in drug discovery and new drug development. Medicinal plants that
have been used in traditional systems of medicine are being studied more than a fine decade
for scientific evaluation and validation of the application, which provides rational and
evidence-based alternative medicines and eventually a backbone of modern day’s herbal drug,
herbal bioactives, semisynthetic and to some extent in synthetic drug and pharmaceutical
industries.
In the light of the foregoing, this Book, "Bioactive Phytochemicals: Drug Discovery to
Product Development" is a tremendously important and in-depth contribution to the subject of
Phyto-pharmaceuticals from drug discovery to product development. The book has brought
together diverse works in the scientific literature under an umbrella on the use of
therapeutically active phytochemicals in the treatment of various chronic disorders. An
excellent effort has been made in this frankly speaking 'a much-needed book' to address the
challenges in plant and plant bioactive based drug development to its transition to the
phytopharmaceutical product comprehensively and sequentially. I am confident that the book
is capable of centering the attention of the reader on the limitless potentials of bioactive
phytochemicals. It does particularly well too in highlighting several current areas of interest
in extraction, isolation, structural characterization, pharmacological screening, product
development and regulatory perspectives of herbal drugs and bioactive phytochemicals.
I have, no doubt, that this book will be well-received (equally) by the researchers in the
phytopharmaceutical industry and academia. Moreover, this book is really good at offering a
vast knowledge on this subject to the chemist and pharmacy students.
Dr. Sohail Akhter
Principal Scientist
New Product development, Global R & D
Teva Pharmaceuticals
Runcorn, United Kingdom
ii
PREFACE
Early man explored natural surroundings such as plants, animals, and minerals to find the
remedies of different ailments. An alternative system of medicines such as Ayurveda, Unani,
Kampo, and traditional Chinese medicinal products are used as dietary supplements and
nutraceuticals reported to become a major part in the treatment of chronic disorders such as
diabetes, cancer, malaria, arthritis, inflammation, liver disorders, and cardiac disorders. WHO
Traditional Medicine Programme is also designed on the potential of ethnomedicine as a lead
for drug discovery. Compounds from plants have been extracted to investigate novel
therapeutics since the origin of medicine. Natural bioactive compounds from medicinal plants
are inexplicably diverse in chemical structure, biological properties and the unmet therapeutic
requirements attribute to develop an interest in the reemergence of bioactive natural
compounds that will lead to isolation, structural characterization and finding pharmaceutical
activity.
The book discusses the scope and approaches of drug discovery from natural products;
cultivation, collection and processing of medicinal plants; Methods and high throughput
techniques for extraction, isolation, and characterization of bioactive phytochemicals;
pharmacological screening for its activity and formulation development as well as quality
control of natural medicinal products. It also discusses the regulations specified for natural
medicinal products in a different region of the world. The last chapter of the book is devoted
to discuss the role of natural herbal products for the treatment of human diseases such as
cancer, cardiovascular diseases, diabetes, obesity, inflammation, and neurological disorders.
Each chapter concludes with a general reference section, which is a bibliographic guide to
more advanced texts.
This book will provide in-depth information and comprehensive discussion on extraction,
isolation, structural characterization, pharmacological screening of bioactive phytochemicals
as well as product development and regulatory perspective of herbal medicine. Researchers,
industries, and students would be interested in this concise body of information. The
contributing authors are drawn from a rich blend of experts in various areas of herbal
medicine encompass drug discovery to product development.
Dr. Javed Ahmad
College of Pharmacy
Najran University
KSA
Co-editor
Javed Ahamad
Department of Pharmacognosy,
Faculty of Pharmacy,
Tishk International University,
Kurdistan Region, Erbil,
Iraq
iii
List of Contributors
Afrin Salma Department of Pharmaceutical Chemistry, Translum Institute of Pharmaceutical
Education and Research, Meerut (UP), India
Ahmed Nawaz Khan School of Pharmacy, Graphic Era Hill University, Dehradun Uttarakhand,
248002, India
Asad Ali Department of Chemistry, School of Chemical and Life Sciences, Jamia
Hamdard University, New Delhi, India
Chandra Kala Faculty of Pharmacy, Maulana Azad University, Jodhpur, Rajasthan, 342802,
India
Dinesh Kumar Patel Department of Pharmaceutical Science, Sam Higginbottom, University of
Agricultural Tech. and Science, India
Esra T. Anwer Department of Pharmaceutics, Faculty of Pharmacy, Tishk International
University, Kurdistan Region, Iraq
Faraat Ali Laboratory Services, Botswana Medicines Regulatory Authority, Plot 112
International Finance Park, Gaborone, Botswana
Govind Prasad Dubey Study Director and Coordinator - Collaborative Program, Institute of Medical
Sciences, Banaras Hindu University, Varansi, India
Jamia Firdous Department of Pharmacy, Institute of Bio-Medical Education and Research,
Mangalayatan University, Aligarh, India
Jaswanth Albert Faculty of Pharmacology, Surabhi Dayakar Rao College of Pharmacy, Gajwel,
Rimmanaguda, Hyderabad, Telangana, India
Javed Ahamad Department of Pharmacognosy, Faculty of Pharmacy, Tishk International
University, Erbil, Kurdistan Region, Iraq
Javed Ahmad Department of Pharmaceutics, College of Pharmacy, Najran University,
Kingdom of Saudi Arabia
Kamna Sharma Department of Pharmaceutical Analysis, Indo-Soviet Friendship College of
Pharmacy, Moga, Punjab, India
Kamran Javed Naquvi Department of Pharmacognosy & Phytochemistry, Faculty of Pharmaceutical
Sciences, Rama University, Rama City, Mandhana, Kanpur (Uttar Pradesh) -
209 217, India
Manisha Trivedi NIMS University, Jaipur, Rajasthan, India
Mohammad Shabib
Akhtar
Department of Clinical Pharmacy, College of Pharmacy, Najran University,
Najran, Kingdom of Saudi Arabia
Muath Sh. Mohammed
Ameen
Department of Pharmaceutics, Faculty of Pharmacy, Tishk International
University, Kurdistan Region, Iraq
Naila Hassan Ali
Alkefai
Department of Pharmacognosy, Faculty of Pharmacy, University of Hafer
Albatin, Hafer Albatin, KSA
Nehal Mohsin Department of Clinical Pharmacy, College of Pharmacy, Najran University,
Najran, Kingdom of Saudi Arabia
Abdul Samad Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Tishk
International University, Kurdistan Region, Iraq
LY
Omji Porwal Department of Pharmacognosy, Faculty of Pharmacy, Tishk International
University, Erbil, Kurdistan Region, Iraq
Raad A Kaskoos Faculty of Pharmacy, Howler Medical University, Kurdistan Region, Iraq
Rahul Tripathi Medpharm Pharmaceutical Ltd., Guilford, United Kingdom
Sachin Singh School of pharmaceutical Sciences, Lovely professional University, Phagwara-
144411, Punjab, India
Saurabh Gupta Department of Pharmacology, Chitkara University, Jansla, Rajpura, Punjab
140401, India
Shaik Khasimbi Department of Pharmaceutical Chemistry, Delhi Institute of Pharmaceutical
Sciences and Research (DIPSAR), Mehrauli-Badarpur Road, PushVihar,
Sector-3, New Delhi, 110017, India
Shankar Katekhaye Center for pharmaceutical Engineering Science University of Barford, Barford-
BD71DP, United Kingdom
Shehla Nasar Mir
Najibullah
Department of Pharmacognosy, Faculty of Pharmacy, King Khalid University,
Abha, KSA
Showkat R. Mir Department of Pharmacognosy, School of Pharmaceutical Education and
Research, Jamia Hamdard, New Delhi, India
Subasini Uthirapathy Department of Pharmacology, Faculty of Pharmacy, Tishk International
University, KRG, Iraq
Tara Fuad Tahir Faculty of Science and Health, Koya University, Kurdistan Region, Iraq
Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 1-13 1
CHAPTER 1
Drug Discovery from Plant Sources: Scope,
Approach and Challenges
Javed Ahmad1 and Javed Ahamad2,*
1 Department of Pharmaceutics, College of Pharmacy, Najran University, Kingdom of Saudi
Arabia
2 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Erbil,
Kurdistan Region, Iraq
Abstract: Medicinal plants are recognized to fulfill human necessities like food,
clothes, shelter and health. The search for eternal health and longevity to seek remedy
to relieve discomfort prompted mankind to develop many ways and means of health
care systems. Traditional medicines were practiced in ancient civilizations for the cure
of ailments. In recent years, natural products play a very important role in drug
discovery for life-threatening ailments like cancer, malaria, diabetes and cardiovascular
problems. Recently, drug discovery from plants for the treatment of cancer gets more
focused and leads to the discovery of novel anticancer drugs such as paclitaxel,
docetaxel, topotecan, irinotecan, vincristine and vinblastine. Drug discovery from
plants is a long and tedious process, and it requires selecting suitable plants, pre-
clinical screening, clinical evaluation and drug approval for marketing. Herbal
medicines obtained from plants are generally considered safe compared to synthetic
drugs, and secondary metabolites obtained from plants have more chemical diversity
and considered superior to synthetic combinatorial chemicals. In this book chapter, we
comprehensively discussed the advantages and role of higher plants in drug discovery,
steps and approaches of drug discovery from higher plants.
Keywords: Approaches of Drug Discovery, Cancer, Drug Discovery, Diabetes,
Malaria, Natural Products, Plants.
1. INTRODUCTION
Since ancient times, plants are recognized to fulfill human necessities like food,
clothes, shelter and remedies for ailments. Since the early days, mankind explored
natural resources and used them as a remedy for the cure of diseases. The
traditional systems of medicine such as Ayurveda, Unani, Chinese medicine and
* Corresponding author Dr. Javed Ahamad: Department of Pharmacognosy, Faculty of Pharmacy, Tishk
International University, Erbil, Kurdistan Region, Iraq; E-mails: jas.hamdard@gmail.com, javed.ahamad@tiu.edu.iq
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
2 Bioactive Phytochemicals: Drug Discovery to Product Development Ahmad and Ahamad
Homeopathy are examples of ways of treatment of human diseases. In ancient
times, the human beings used natural resources for various purposes, such as food
and medicinal agents [1]. Traditional medicines were practiced in ancient
civilizations; the traditional practitioner gathered knowledge from generation to
generation, and sometimes they document this knowledge. Ancient Indian
scholars such as Charaka and Sushrutha, examined and classified medicinal herbs
based on their properties and they called it as “Gunas”. Charaka arranged 50
groups containing ten herbs in each group, and according to him, these herbs are
sufficient to cure all types of human diseases. Sushrutha also prepared seven
groups from 760 herbs based upon some common properties [2]. Chinese scholars
also worked meticulously and developed remedies from natural resources. One of
the oldest known literature by Pent Sao documented by ShenNung around 3000
B.C., and it contained 365 drugs, one for each day of the year [3]. Hippocrates
“Father of medicine” (460-360 B.C.), Aristotle (384-322 B.C.), Dioscorides (40-
80 A.D.), Galen (131-200 A.D.) and the early Arabian physicians like Rhazes
(865-925 AD) and Avicenna (980-1037 AD) were recognized as pioneers in drug
discovery from natural products [4].
Higher plants have advantages in drug discovery compared to synthetic drugs
because they have a long history of use by humans as food and spices. Natural
products obtained from plants are generally considered safe compared to synthetic
drugs, and the secondary metabolites obtained from plants have more chemical
diversity and considered superior to synthetic combinatorial chemicals [5].
Several natural products recently showed cytotoxic and antitumor activities in
clinical trials and were successfully developed as anticancer drugs such as
vincristine, vinblastine, paclitaxel and docetaxeletc. Drug discovery from plants
today is an expensive and lengthy process. The drug discovery from plants
requires a team effort consisting of experts from different disciplines such as
pharmacognosists, pharmacologists, medicinal chemist and pharmaceutics.
Phytochemicals from medicinal plants serve as a lead compound in drug
discovery and are further used for synthetic or semi-synthetic drug development
to ensure patent protection [6]. In ancient civilization, plants were used as
medicinal agents for the treatment of various human ailments. The knowledge of
medicinal values of these plants was inherited from generation to generation.
Most of these traditional practitioners formulate and dispense their own
medicines; hence this requires proper documentation and research to use natural
products as therapeutically effective and safe medicinal agents [7]. In recent
years, traditional medicines have emerged as an alternative treatment of chronic
diseases and lifestyle disorders such as cancer, malaria, tuberculosis, diabetes,
obesity and cardiovascular complications [8]. Drug discovery from the plant is a
long and tedious process, and it requires selecting suitable plants, biological or
pre-clinical screening, clinical evaluation and drug approval for marketing. In this
Plant Sources Bioactive Phytochemicals: Drug Discovery to Product Development 3
book chapter, we comprehensively discussed the advantages and role of higher
plants in drug discovery, steps and approaches of drug discovery from higher
plants.
2. ADVANTAGES OF DRUG DISCOVERY FROM NATURAL
RESOURCES
Newman and Cragg, reported that totally 1562 new drugs approved during 1981
to 2014, of which, those of plant origin comprised as follows: natural products
(4%; N), derivatives of natural products (21%, ND), synthetic compounds with
natural product-derived pharmacophores (10%; S*/NM), and synthetic drug with
NP pharmacophore (11%; S/NM). It is clear from the above report that natural
products contribute a lot in drug discovery [9]. Plants are considered as equal or
superior in drug discovery because of their chemical diversity and human friendly
in nature based on their long history of use as food by a human. Phytochemicals
have been elaborated within living systems, they are often perceived as showing
more drug-likeness and biological friendliness or tolerance than synthetic
molecules, making bioactive phytochemicals good candidates for further drug
discovery and development [6]. The data on new drugs approved during 1981-
2014 is summarized in Table 1 and Fig. (1).
Table 1. New drugs approved during 1981-2014 from all sources (number = 1562).
Code Category Numbers Percentage
B Biological drugs 250 16%
N Natural drugs 67 4%
NB Botanical drugs with defined mixture 9 1%
ND Natural product derivatives 320 21%
S Synthetic drugs 420 27%
S/NM Synthetic drug with NP pharmacophore 172 11%
S* Synthetic drugs which mimics natural product 61 4%
S*/NM Synthetic compounds with natural product-derived pharmacophores 162 10%
V Vaccine 101 6%
T Total 1562 100%
14 Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 14-30
CHAPTER 2
Cultivation, Collection and Processing of Medicinal
Plants
Omji Porwal1,*, Sachin Kumar Singh2, Dinesh Kumar Patel3, Saurabh
Gupta4, Rahul Tripathi5 and Shankar Katekhaye6
* Corresponding author Omji Porwal: Department of Pharmacognosy, Faculty of pharmacy, Tishk
International University, Erbil, Kurdistan Region, Iraq; Email:omji.porwal@tiu.edu.iq
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
1 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Erbil,
Kurdistan, Iraq
2 School of Pharmaceutical Sciences, Lovely Professional University, Phagwara-144411, Punjab,
India
3 Faculty of Health Science, Shalom Institute of Health and Allied Sciences Sam Higginbottom
University of Agriculture, Technology and Sciences (SHUATS)-State University (Formerly
Allahabad Agriculture Institute) Naini, Allahabad, India
4 Chitkara College of Pharmacy, Chitkara University, Chandigarh-patiala highway (NH-4),
Rajpura Punjab 140401-India
5 Medpharm Pharmaceutical Ltd. Guilford, United Kingdom
6 Center for pharmaceutical Engineering Science University of Barford, Barford-BD71DP, United
Kingdom
Abstract: Nature has provided us better surroundings for the expansion and
development of medicinal plants for thousands of years. Medicinal values of plants
date back to ancient times on belief of its safety and economic value. Even in today’s
scenario, about 80% of the globe population primarily depends on the alternative
system of medicines for their foremost healthcare requisite. Plants contain different
types of secondary metabolites also called bioactive components of the plants
responsible for their medicinal value in nature. Scientific cultivation permits the
applying of contemporary technological aspects like mutation, polyploidy and
hybridization for the development of a better amount of secondary metabolites from the
plants and their byproducts. Cultivation, collection and preservations of medicinal
plants need simple techniques that preserve the medicinal values of natural products.
Pharmacological activities of medicinal and aromatic plants are mainly depending upon
the presence of various bioactive phytochemicals like alkaloids, glycosides, tannins,
resins, volatile oil, etc. Growth and development of medicinal plants and their
secondary metabolites are mainly influenced by the physical surroundings, sunlight,
temperature, rainfall, and nature of the soil. Seasonal variation and geographical region
can also affect the quality of medicinal and aromatic plants as the concentration of
bioactive constituent could be changed through natural factors. In the present book
Medicinal Plants Bioactive Phytochemicals: Drug Discovery to Product Development 15
chapter, we have discussed all the important information needed for the cultivation,
collection and processing of medicinal and aromatic plants that affect the quality of
medicinal plants.
Keywords : Cultivation, Collection, Drying, Harvesting, Medicinal Plants, Plant
Growth Hormones.
1. IMPORTANCE OF MEDICINAL PLANTS
The crude drug that reaches the market and pharmaceutical industry must have
some basic characteristics and quantity of active constituents for their claim
therapeutic potential. Cultivation produces a quality of plants, increases secondary
metabolite concentration, generates hybrids that produce specific
phytoconstituents, ensures regular supply of crude drugs and also leads
industrialization, etc. The gathering of crude medicine from cultivated plants
provides a higher yield and therapeutic quality. Medicinal plants require sunny,
aerated places sheltered from gale and late winter frosts. The soil must be fertile
and consist of the required amounts of Na, P, Cu, alloy, organic and other
essential elements for the better growth of medicinal and aromatic plants.
Continuation of the farming structure has become a vital affair throughout the
globe. A large number of the viable matter is related to the quality and time-
dependent changes in the soil [1]. It is well known that in-depth cultivation has
led to a swift rebuff in organic matter and nutrient levels besides affecting the
physical things of clay [1]. However, the management practices with organic
materials influence agricultural properties by the physical, chemical and
biological properties of clay [2]. The implementation of organic amendments has
long been recognized as an efficient suggestion for the improvement in the
structure and fertility of the soil [3]. Increasing the microorganism diversity,
activity, population and moisture-holding capability of soils also improve crop
yield [4].
2. CULTIVATION OF MEDICINAL PLANTS
The demand for medicinal and aromatic plants has been increased in the market
due to its better medicinal value and pharmacological activity that’s why we need
better care of medical care and mechanical management. If no scientific method
of cultivation is available, then ancient strategies of cultivation should be
followed. Otherwise, there is a need to develop a way of cultivation through
scientific studies. The principles of agriculture practices together with appropriate
rotation step with environmental suitableness, tailored to plant cultivation as per
source needs. Conversation Agriculture (CA) capability ought to become behind
16 Bioactive Phytochemicals: Drug Discovery to Product Development Porwal et al.
where applicable, exceptionally builds up of fertile matter and conversation of soil
humidness.
2.1. Method of Propagation
2.1.1. Sexual or Seed Propagation
Cultivation of medicinal plants by sexual propagation is by spreading or sowing
seeds in suitable soil. Seeds are either shown by the dibbling method, in which the
seeds of average size and weight are available, they are sown by placing in holes
eg, fennel, coriander, etc., or by the broadcasting method and this method is more
applicable in case of the small size seeds. In this method, the seeds are scattered
freely in well-prepared soil for cultivation, e.g. Isapgol linseed and sesame.
2.1.2. Asexual or Vegetative Propagation
In the case of the asexual method of vegetative propagation, the vegetative part of
a plant, such as a stem or a root, is placed in such an environment that it develops
into a new plant. The advantages of asexual propagation are there is no variation
between the plant grown and plant from which it is grown. Seedlings of fruits can
only be propagated vegetatively (e.g. Grapes, pomegranates and lemon), plant
start bearing earlier as compared to seedling trees, budding or grafting encourages
disease resistance varieties of plants, and inferior varieties can be overlooked.
Vegetative propagation is done by sowing various parts of the plants in well-
prepared soil. The following are examples of vegetative propagation:
Bulbs: squill, garlic
Corms: colchicum, saffron
Tubers: jalap, potato
Rhizomes: ginger, turmeric
Runners: peppermint
Suckers: mint, pineapple, banana, etc.
3. FACTORS AFFECTING CULTIVATION
Farming of medicinal and aromatic plants takes cognizance of plant habits and
climatic requirements for their favourable growth. The factor which affects the
Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 31-53 31
CHAPTER 3
Extraction of Bioactive Phytochemicals
Javed Ahamad1,*, Naila Hassan Ali Alkefai2 and Shehla Nasar Mir Najibullah3
1 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Kurdistan
Region, Erbil, Iraq
2 Department of Pharmacognosy,Faculty of Pharmacy, University of Hafer Albatin, Hafer Albatin,
KSA
3 Department of Pharmacognosy, Faculty of Pharmacy, King Khalid University, Abha, KSA
Abstract: The success of pharmacological or analytical study depends upon proper
choice of the extraction method. As extraction process is the starting step in the
evaluation of any herbal drug, hence it should be designed in such a way that the
extracts contain maximum amount of desired secondary metabolites. Several classical
extraction methods are used since ancient times, such as percolation, digestion,
decoctions, and maceration. Due to the advancement in technology, several extraction
techniques have been developed, such as ultrasound assisted extract, microwave
assisted extraction, pressurized liquid extraction, supercritical fluid extract and enzyme
assisted extraction. These extraction techniques provide samples for qualitative and
quantitative analysis of the natural product. The extracts obtained from these extraction
methods are also used for the isolation and characterization of bioactive natural
products. This book chapter provides a comprehensive overview of a variety of
classical and modern extraction methods used in drug discovery and development from
the natural product.
1. INTRODUCTION
Extraction is the first step in obtaining desired bioactive phytochemicals from
medicinal plants and it is defined as“the separation medicinally active portions of
plant and animal tissue using selective solvents by standard procedures”. Medi-
cinal plants and their bioactive natural compounds have records of use as food,
cosmetics and medicine.
* Corresponding author Dr. Javed Ahamad: Department of Pharmacognosy, Faculty of Pharmacy, Tishk
International University, Kurdistan Region, Erbil, Iraq; E-mails: jas.hamdard@gmail.com & javed.ahamad@tiu.edu.iq
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
Keywords: Bioactive Phytochemicals, Extraction, Isolation, Modern Extraction
Methods, Natural Products, Secondary Metabolites.
32 Bioactive Phytochemicals: Drug Discovery to Product Development Ahamad et al.
More advanced and efficient extraction and separation techniques are required due
to the increased demands of these phytochemicals from the industries. Many
traditional or classical extraction methods have been used previously to extract
bioactive phytochemicals such as maceration, percolation, digestion decoction,
etc. In recent decades, certain modern extraction techniques have also been
developed for the efficient extraction of natural products due to advances in
technology. Even the advent of modern and sophisticated extraction techniques
has not yet been found to be suitable for all forms of phytochemicals [1]. That is
why, depending on the nature of crude drugs and the suitability of extraction
techniques, we used specific methods for particular crude drugs [2, 3]. Medicinal
plants are the largest source of bioactive phytochemicals in several conventional
and modern medical practices, which are used traditionally as medicines.
Medicinal plants containing essential oils are a major source of fragrances,
flavors, cosmeceuticals and health beverages. Elevated temperature causes a
detrimental effect on the chemical composition of essential oils for extraction.
That is why special extraction methods are used to separate these compounds
under low temperatures. Mostly medicinal plants are traded as such in bulk from
many developing and underdeveloped countries for value-added in developed
countries. The first step in adding value to the wealth of medicinal plants is to
extract in the form of galenicals or dried extracts. Various types of herbal extracts,
infusions, tinctures and herbal tea are commercially available in the market [4].
These preparations are also known as decoctions, fluid extracts, pilular extracts or
powdered extract. The need to develop a standardized extraction procedure for
crude drugs is to achieve the therapeutically desired portions and to eliminate
unwanted materials by using a selective solvent treatment. The extracts thus
obtained after standardized methods of extraction could be used as medicinal
agents as such or incorporated in the finished dosage form such as tablets and
capsules or used in further studies [5].
These extracts contain bioactive phytochemicals of medicinal plants belonging to
different classes of secondary metabolites such as alkaloids, glycosides,
terpenoids, flavonoids, tannins, resins, fixed oils, essential oils and lignans etc. In
order to be used as modern drugs, extracts could be further processed by means of
various isolation and purification techniques of bioactive phytochemicals such as
counter current chromatography, flash chromatography, preparative HPLC,
preparative TLC and preparative GC etc. The book chapter provides a
comprehensive overview of a number of classical and modern extraction methods
that are used in the production and development of herbal drugs.
Bioactive Phytochemicals Bioactive Phytochemicals: Drug Discovery to Product Development 33
2. STEPS INVOLVED IN THE EXTRACTION OF MEDICINAL PLANTS
Numerous new methods have been developed along with conventional methods,
but so far, no single method is considered the standard for plant extraction of
bioactive compounds. The qualitative and quantitative studies of bioactive
compounds from plant materials depend mostly on choosing the proper method of
extraction. The various steps involve the extraction of medicinal ingredients from
plant materials, such as drying and grinding of plant materials, selection of
suitable solvents, extraction, filtration, concentration and drying of the extract [6].
2.1. Drying and Grinding of Plant Materials
In most cases, before the place of work, plant material is dried in the atmosphere.
It can be dried at room temperature or below 45 °C in the oven. If plant materials
were left to stand for several days, compacted samples of fresh plant material with
little air circulation could experience fungal infestation and fermentation at
elevated temperatures. Well ventilated places and homogeneous material
distribution should therefore be ensured. The equipment used for grinding plant
materials is advanced mills, blenders, simple axes, scissors, grinders, or knives.
When extracting thermolabile or volatile phytocompounds the milling stage may
be omitted to avoid heat-generating losses during communition. The grinding
process assists solvent penetration into the cellular structure of the plant tissues,
thus helping to dissolve the secondary metabolites and increase extraction yields.
Size reduction also maximizes the surface area, which in effect facilitates the
transfer of mass of the active principle from the plant material to the solvent.
2.2. Selection of Suitable Solvents
Care should be taken to consider the solvents selected for extraction. The selected
solvent should dissolve the under-study secondary metabolites, be easy to remove,
and be inert, non-toxic and, not easily flammable. Solvents should be distilled if
they are of low or unknown quality before use. Examples of solvents commonly
used for extraction studies include water, ethanol, and methanol etc.
2.3. Extraction Process
The simplest extraction processes employed are organic solvent extraction;
percolation, maceration, and extraction using a Soxhlet device; and water
extraction; infusion, decoction, and distillation of steam. Extraction of plant
materials nowadays by various modern or non-conventional techniques of
54 Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 54-72
CHAPTER 4
Isolation and Purification of Bioactive
Phytochemicals
1 Department of Pharmacognosy & Phytochemistry, Faculty of Pharmaceutical Sciences, Rama
University, Rama City, Mandhana, Kanpur (Uttar Pradesh) - 209 217, India
2 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Erbil,
Kurdistan Region, Iraq
3 Faculty of Pharmacy, Howler Medical University, Kurdistan Region, Iraq
4 Department of Pharmacognosy, Faculty of Pharmacy, University of Hafer Albatin, Hafer
Albatin, KSA
5 Department of Pharmaceutical Chemistry, Translum Institute of Pharmaceutical Education and
Research, Meerut (UP), India
6 Department of Pharmacognosy, School of Pharmaceutical Education and Research, Jamia
Hamdard, New Delhi, India
Abstract: Various forms of natural products such as plant extracts, pure
phytochemicals and herbal formulations containing natural products offer tremendous
opportunities for new drug discoveries and the credit goes to its chemical diversity.
Since time immemorial medicinal plants in its various forms and have been used to
treat chronic diseases such as malaria, tuberculosis, cardiovascular diseases, etc.
Recently along with the crude form of medicinal plants and their isolated active
principles are also being used to cure several maladies. Isolation of bioactive
constituents from the medicinal plants has always been a challenge because of the
complexities involved in separation process, but the recent technological advancement
in this field has facilitated the isolation process of chemical constituents from the
plants. This book chapter offers a comprehensive review on the procedural techniques
and application of classical column chromatography, prep-TLC, modern isolation and
purification techniques such as flash chromatography, prep-HPLC, prep-GC, counter
current chromatography, etc.
* Corresponding author Dr. Kamran Javed Naquvi: Department of Pharmacognosy & Phytochemistry, Faculty of
Pharmaceutical Sciences, Rama University, Rama City, Mandhana, Kanpur (Uttar Pradesh) - 209 217, India; E-mail:
kjnaquvi@gmail.com
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
Kamran Javed Naquvi1,*, Javed Ahamad2, Raad A Kaskoos3, Naila Hasan Ali
Alkefai4, Afrin Salma5 and Showkat R. Mir6
Keywords: Column Chromatography, Counter-Current Chromatography, Flash
Chromatography, Isolation, Natural Products, Phytochemicals, Preparative-GC,
Preparative-HPLC, Preparative-TLC, Purification.
Isolation and Purification Bioactive Phytochemicals: Drug Discovery to Product Development 55
1. INTRODUCTION
Medicinal plants have been reported to be repository of various types of bioactive
compounds, which possessing tremendous therapeutic properties. The use of
plants as a therapeutic agent over a very long time period has been established.
The magnificent and diverse plant kingdom is well known for its medicinal
values. Herbal therapies across the globe for various diseases are largely
dependent on the therapeutic potential of medicinal plants [1]. Plants are being
used for medicinal purposes long before the prehistoric era. Since ancient times
scholars from Egypt, India, China and Arabian countries compiled and utilized
herbs as medicinal agents and fulfilled their basic health care needs from these
medicinal herbs. Herbal therapies have been considered as the safest one because
of their minimal or no side effect characteristics. As these remedies do align with
the nature and hence offer the biggest advantage. One of the facts relating to
herbal treatments is that it is independent of any age group and the sexes. Natural
flora is considered as one of the richest and biodiverse resource of active
ingredients, which can be used for pharmacopeial, non-pharmacopeial or synthetic
drug development. Moreover, some plants also possess nutritional values along
with their therapeutic potential and hence they are the most recommended ones in
the therapeutic domain [2].
Drug discovery and development is a complex, time consuming and very
expensive process. Drug discovery process involves the identification of new
chemical entities (NCES), which should possess the desired pharmacokinetic and
pharmacodynamic properties. The NCES are generally obtained through isolation
from natural resources or through chemical syntheses. In the pre genomic era and
when there was no high throughput screening (HTS), more than 80% of drug
substances were plant based or were the derivatives of plant based medicines [3,
4]. Newman and Cragg, reported that total 1562 new drugs were approved during
1981 to 2014, out of which natural products (4%; N), derivatives of natural
products (21%, ND), synthetic compounds with natural product-derived
pharmacophores (10%; S*/NM), and synthetic drug with NP pharmacophore
(11%; S/NM). From the above report it is clear that about 51% of the drugs
discovered come directly or indirectly from natural products [5].
WHO estimated about 20,000 plants having medicinal values exist in 91 countries
including 12 mega biodiverse countries. Natural products isolated from plants
offer unparalleled source of chemical diversity for the discovery of biologically
active scaffolds [6]. The critical steps for revelation of the biologically active
compounds from plant resources are extraction, isolation, characterization and
pharmacological screening of bioactive compounds followed by toxicological and
clinical evaluation. Considering the fact that plant extract happened to possess a
56 Bioactive Phytochemicals: Drug Discovery to Product Development Naquvi et al.
combination of various types of bioactive compounds or the plant constituents
with different polarities, their separation still poses a great challenge for the
process of identification and characterization of bioactive compounds [7].
Isolation of single constituents from plant generally involves separation of
components from its mixture (extract). The separated single constituents might
contain impurities which are further separated by several purification processes.
Purification refers to the process of separating or extracting the target compound
from other (possibly structurally related) compounds or contaminants. Generally
chromatographic methods are employed for separation and purification of
bioactive chemicals from plants. For isolation of single component mostly column
chromatography, prep-TLC, counter-current chromatography is applied and for
purification, prep-HPLC, MPLC is used [8 - 10]. Isolation and purification of the
bioactive phytochemicals from medicinal plants have always posed the problems
associated with their chemical complexities. Recent technological advancements
related to isolation and separation techniques have made the process quite easier
than before [11]. For the isolation and purification purpose, various solvents with
differential polarities either individually or in combination are used. Bioactive
guided fractionation and isolation of phytochemicals are now practiced widely.
Preparative thin layer chromatography (prep-TLC) and column chromatography
are the oldest chromatographic methods applied for isolation and purification of
bioactive phytochemicals from plant sources. These traditional methods are still
the first choice due to their convenience with respect to the economic feasibility
and availability in various stationary phases [12]. Recent advancement in isolation
and purification techniques leads to develop more sophisticated instruments such
as preparative high pressured liquid chromatography (prep-HPLC), countercurrent
chromatography (CCC), and medium pressure liquid chromatography (MPLC)
which expedites the process of isolation and purification of bioactive compounds
[13]. In this book chapter we have compiled a comprehensive overlook about
isolation and purification techniques such as prep-TLC, prep-HPLC, CCC, MPLC
and preparative gas chromatography (prep-GC).
2. TECHNIQUES OF NATURAL PRODUCT ISOLATION
2.1. Counter-Current Chromatography (CCC)
Natural product extracts contain mixtures of highly complex compounds; because
of this reason the purification process becomes a highly challenging task.
Purification protocols sometimes take months or years, as it involves isolation of a
single active molecule from the hundreds of compounds that can be present in the
complex mixture form in an extract. Thus it is critical for a phytochemist to
Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 73-99 73
CHAPTER 5
Spectroscopic Techniques for the Structural
Characterization of Bioactive Phytochemicals
Showkat R. Mir1,*, Tara Fuad Tahir2, Javed Ahamad3, Raad A Kaskoos4,
Naila Hassan Ali Alkefai5 and Abdul Samad6
1 Department of Pharmacognosy, School of Pharmaceutical Education and Research, Jamia
Hamdard, New Delhi, India
2 Faculty of Science and Health, Koya University, Kurdistan Region, Iraq
3 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Kurdistan
Region, Iraq
4 Faculty of Pharmacy, Howler Medical University, Kurdistan Region, Iraq
5 Collage of Pharmacy Department of Pharmacognosy & Phytochemistry University of Hafer
Albatin Hafer Albatin, Kingdom of Saudi Arabia
6 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Tishk International University,
Kurdistan Region, Iraq
Abstract: This chapter deals with the structural elucidation of natural products using
UV-visible, FT-IR, NMR (1D and 2D) spectroscopy and Mass spectrometry. Key
concepts associated with these techniques are introduced here. Various spectra of
natural compounds have been included with a brief discussion of the functional groups
and structural features that they reveal. The use of these spectroscopic techniques in
drug discovery from natural products has been reviewed, highlighting the advantages
of these techniques. The importance of each technique is discussed with suitable
examples of natural products obtained from medicinal plants. Some of these
techniques, such as FT-IR and NMR spectroscopy, are primarily used for identification
purposes; while UV-visible spectroscopy and mass spectrometry are used for the
purpose of analysis and structural elucidation. Recent advancements in the isolation
techniques such as counter-current chromatography, supercritical fluid
chromatography, preparative high-performance liquid chromatography (prep-HPLC),
and preparative gas chromatography (prep-GC) have made the availability of novel
natural compounds from plants possible for their structural elucidation and biological
screening. Development of hyphenated techniques such as LC-NMR, UPLC-MS, and
GC-MS have made simultaneous isolation and structural elucidation of natural
products achievable. In this chapter, we have summarized basic principles of UV-
visible, FT-IR, 1D and 2D NMR spectroscopy and mass spectrometry and their role in
the determination of structures of natural compounds citing suitable examples.
* Corresponding author Showkat R. Mir: Department of Pharmacognosy, School of Pharmaceutical
Education and Research, Jamia Hamdard, PO Hamdard Nagar, New Delhi, India 110 062;
E-mail: showkatrmir@gmail.com
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
74 Bioactive Phytochemicals: Drug Discovery to Product Development Mir et al.
1. INTRODUCTION
Phytochemistry deals with the study of various forms of secondary metabolites
that are elaborated and accumulated in plants and also relate to their biosynthesis,
turnover, metabolism, correlation and biological functions. Their isolation and
characterization are important aspects of phytochemical studies [1]. Plant
secondary metabolites are not only used as crude, raw and unmodified drugs, but
they are also regarded to provide templates and starting materials for the
production of various semi-synthetic drugs. For instance, diosgenin from
Dioscorea species can be synthetically transformed to different steroids having
anabolic, anti-inflammatory and oral contraceptive properties [2]. Phytochemicals
that are used as therapeutic agents belong to classes such as alkaloids, glycosides,
flavonoids, iridoids, terpenoids, resins, tannins, lignans, lipids, etc [3 - 7]. In
recent decades, phytochemistry has played a central role in drug discovery and
development processes. The enormous success of taxol has encouraged the
scientific community worldwide to take up the research on drug discovery and
development from plants. Recent technological advancements have opened
various possibilities of phytochemical research to attain the new chemical entities
as the drugs of future. The outlook of natural product chemistry has completely
changed in the last few decades, which has witnessed explosive growth because of
the advancement in extraction, isolation and characterization techniques. This has
transformed the pace of drug discovery and development from natural substances.
Development of separation techniques such as medium pressure liquid
chromatography, preparative high-performance liquid chromatography (prep-
HPLC), droplet counter-current chromatography (DCCC) and preparative gas
chromatography (prep-GC), etc., have greatly expedited and simplified the
isolation and separation of complex mixtures of plant constituents that were
thought to be inseparable earlier. Discovery of the powerful hyphenated
techniques such as LC-NMR, LC-MS, and GCMS-MS has made the simultaneous
isolation and structure elucidation of natural products possible [8, 9]. This chapter
gives a preliminary overview of structural elucidation of natural products using
UV-visible, FTIR, 1D and 2D NMR spectroscopy and Mass spectrometry.
2. STRUCTURAL ELUCIDATION OF PHYTOCONSTITUENTS
The step that precedes structural elucidation of natural compounds is their
isolation and purification. Recent advancements in the isolation and purification
techniques have made the process easy and fast. Soxhlet apparatus, Clevenger
Keywords: FT-IR Spectroscopy, Mass Spectrometry, Natural Products, NMR
Spectroscopy, UV-visible Spectroscopy, Structural Characterization.
Spectroscopic Techniques Bioactive Phytochemicals: Drug Discovery to Product Development 75
apparatus, supercritical fluid extractor, etc. are used for the separation of a
specific mixture of phytochemicals from their matrices. It is followed by
purification by chromatographic methods, liquid-liquid extraction, fractional
distillation, solid-phase extraction, etc. These methods can be used repeatedly or
in combination to attain required purity. Preliminary phytochemical screening
consisting of physicochemical tests may also be employed to ascertain the nature
of these compounds.
The preliminary characterization of phytochemicals is carried out, preferably by
TLC analysis. If it yields a single spot on TLC plates in different mobile phases, it
is considered fit for further characterization studies. Other parameters may also be
used depending upon the nature of the isolates such as sharpness of melting/
boiling/ freezing point, refractive index, optical density, etc. Once the results of
these preliminary studies show that the chemical compound is of desired purity
and belongs to a specific class of phytoconstituents, the advanced techniques of
structural determination are applied, such as UV, IR, NMR spectroscopy and
mass spectrometry.
An already known compound can be identified by matching it's spectra with the
reference spectra available in different libraries or databases. If the direct
comparison can not be made, the spectra of related compounds reported earlier
can be used for judicious structural comparison. On the other hand, for the
characterization of a novel or an unknown compound, an extensive spectral
analyses is performed [10 - 12].
2.1. UV-Visible Spectroscopy
Ultraviolet and visible spectroscopy is a technique to determine the absorption
spectrum of a compound within a wavelength range of 200 to 800 nm. In this UV-
visible region, a molecule absorbs the energy and undergoes through the changes
in the electronic energy levels. Various chromophores within a molecule lead to
the generation of a characteristic absorption spectrum. The absorption pattern of
colourless solutions is measured in the range of 200 to 400 nm, while the coloured
solutions are subjected to colourimetry in the range of 400 to 800 nm [13]. The
spectra record wavelength of maximum and minimum absorption, as well as the
intensity of absorption at a particular wavelength. For the identification purposes
of many plant constituents, the UV-visible spectrum plays a very crucial role. The
absorption spectra are critically valuable for the identification and determination
of the chemical structure and analysis of natural compounds [14].
The electronic transitions in the molecules are classified as per the participating
molecular orbitals. From the four possible transitions (n to π*, π to π*, n to σ, σ to
100 Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 100-120
CHAPTER 6
Pharmacological Evaluation of Herbal Medicine
Subasini Uthirapathy1,*, Javed Ahamad, Jaswanth Albert3 and Govind
Prasad Dubey4,*
Abstract: Natural products broadly incorporate plant-based product as well as marine,
plants, microorganisms and minerals. There are several animal products that are also
consumed by humans as a food health supplement and also medicine. Herbal products
have a variety of phytocompounds. . The identification of these molecules is essential
for standardization and quality control of the herbal products. The world wide variety
of plants and their species have been identified and i.e. 250000. Morphological and
chemical constituents of plant species vary from one ethnic group to another.
Humidity, temperature and altitude soil conditions are all responsible for both
structural and functional characteristics of the natural products. Therefore
standardization and quality control are the big challenges before using natural products.
Heavy metal toxicity, microbial load and aflotoxin are the major and important
parameters of evaluation of the safety profile of the natural products. Therefore, before
using the natural product, we should characterized the structural variation to establish
structural and functional relationships. In the present paper, some of the experimental
models have been described, which will be useful in the preparation of standard plant
based products.In order to provide standard product, some important experimental
methods have been described in this paper. Herbal medicinal products have a wide
scope of assorted variety of multidimensional synthetic structures; in the ongoing
occasions, the utility of normal items as natural capacity modifiers has impressive
consideration. The botanical study evaluated the identification of 250,000 to 350,000
plant species over the planet. Notwithstanding, only around 35,000 species have been
utilized in various networks of the world for the treatment of different infirmities.
Nonetheless, this exceptional fortune needs exhaustive consideration as far as
biological and pharmacological screening to serve humankind against different
diseases. The traditional system of treatment, varying in idea and convention,
represents well-created frameworks, for example, Allopathic, Homeopathic, Ayurvedic
* Corresponding author Dr. Govind Prasad Dubey and Subasini Uthirapathy: Institute of Medical
Sciences, Banaras Hindu University, Varanasi, India; Tel: +0091 9450 9639 42 and Department of
Pharmacology, Faculty of Pharmacy, Tishk International University, KRG, Iraq;
E-mails: gpdubey13@gmail.com & subasini.uthirapathy@tiu.edu.iq
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
1 Department of Pharmacology, Faculty of Pharmacy, Tishk International University, KRG, Iraq
2 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, KRG, Iraq
3 Faculty of Pharmacology, Surabhi Dayakar Rao College of Pharmacy, Gajwel, Rimmanaguda,
Hyderabad, Telangana, India
4 Institute of Medical Sciences, Banaras Hindu University, Varanasi, India
Herbal Medicine Bioactive Phytochemicals: Drug Discovery to Product Development 101
and Chinese system of treatment. A large portion of the enlightened countries has built
up their own Materia Medica, assembling insights concerning different plants utilized
for remedial purposes. This chapteris focused on the diverse pharmacological screening
techniques with point by point illustration of genuine biological investigations that are
useful to discover new bioactive phytochemicals in antidiabetic, analgesic,anti-
inflammatory, neuroprotective, anti-obesity, and depression disorders and so on.
Preclinical study is strictly required to follow CPCSEA guideline into the
pharmacological practical.This chapter provides essential knowledge of practical
features of the experimental Screening pharmacology right from laboratory animal
handling, the important techniques and methodology used in experimental
pharmacology. The experimental procedure described in this chapter is planned on the
basis of strong technical materials and personal knowledge in hands-on experiments
under the guidance of renowned personalities. This chapter is arranged to understand
the experimental animal handling techniques by witnessing the induction experiments.
These experimental procedures will be helpful for graduates and postgraduates students
associated with pharmacology, toxicology and researcher. Simple and newer animal
models have been combined, which may help the students to occupy in new drug
development activities. Also, some important points have been conferred e.g. ethics of
animal experimentation, blood collection techniques, euthanasia, animal care and
handling.
1. INTRODUCTION
Standardization of natural products is complex and challenging. It requires a
multidisciplinary approach to prove the therapeutic value of plant-based medicine
in comparison to synthetic chemicals. Plant-based natural products are used for
the centuries. The traditional knowledge which is a practice in the form of
Ayurveda, Siddha, Unani and Chinese medicine. Generally, the traditional form
of medicine utilizes multiple herbs so poly-herbal and poly-molecular formulation
require providing pharmacological action, bioavailability and bioequivalence
study to compare the risk-benefit ratio in comparison to synthetic chemicals. The
approach of the above system of traditional medicine is holistic. Therefore, to
meet the national and international regulatory norms, several guidelines have been
framed by different countries. The universal acceptance of traditional medicine
requires-complying regulatory norms. In the present book chapter, we have
described the experimental model required in the including toxico-kinetic and
toxicodynamic effect to provide the scientific basis of natural products.
Medication discovery is prompting to be a challenging logical undertaking to
discover strong and suitable lead applicants, which is only the procedure stream
from a screening of regular items to another disengage that requires ability and
experience. It portrays the bioactive compounds obtained from natural resources,
Keywords: Anti-inflammatory Activity, Antidiabetic Activity, Anti-Obesity,
Bio-Logical Approach, Drug Discovery, Herbal Medicinal Products, Neurop-
harmacology, Pharmacological Evaluation.
102 Bioactive Phytochemicals: Drug Discovery to Product Development Uthirapathy et al.
its phytochemical examination, characterization and pharmacological
examination. The trial on animals gives a general thought regarding the tested
medication (pharmacodynamics, pharmacokinetic and toxicology) through
different strategies engaged with the procedure. In any case, a definitive objective
of any medication research is to utilize the medication in the patient’s care [1].
The preclinical study is principally intended to discover a lead compound with
wanting adequacy and well-being for clinical study through pharmacokinetics and
pharmacodynamics information acquired from in vitro and in vivo investigations.
Pharmacology is a therapeutic science that structures a spine of the restorative
calling as medications structure the foundation of treatment in human ailments.
Consequently, it is of most extreme significance to portray the pharmacological
basis of therapeutics to expand the advantages and limit the dangers of
medications to beneficiaries [2].
Broadly experimental animals are divided into three categories such as Rodents
(Mouse, Rat, Guinea pig, black mice, Hamster etc.), non-Rodents (Rabbit, Dog,
Cat, Monkey, Pig etc.) and miscellaneous (Frog, Pigeon, Zebrafish, Chicken). The
proper selection of animal models is one of the paramount steps in the evaluation
of new drugs pharmacologically.The animal model preferred for the study must
be producing a similar disease profile as in the human. Once preclinical is
approved, it can be conducted under CPCSEA (Committee for the Purpose of
Control and Supervision of Experiments in Animals) guidelines.Bioassays are
designed to check the pharmacological activity or efficacy of a new drug, to find
the role of endogenous ligands and to find drug toxicity [3]. This book chapter
note on pharmacology is principally a note for undergraduate wellbeing science
students, for example, wellbeing official, nursing, birthing assistance and lab
innovation students. In any case, other wellbeing experts whose career includes
drug treatment or related viewpoints ought to likewise discover a significant part
of the material applicable. The objective of this book chapter is to find out the
therapeutic effect; to study the toxicity, and; to study the mechanism and site of
action of drugs. Bioassay is the process of evaluation of the drug biologically. In
this chapter, we discussed common laboratory animals used pharmacological
screening, ethics of animal experimentation, and pharmacological screening
methods of major human diseases.
2. COMMON LABORATORY ANIMALS
2.1. Albino Rats (Rattus Norvegicus)
Two strains were commonly used: (a)Wistar Rats and (b) Sprague Dawley Rats.
The normal weight of these rats ranges from 150-300g. The main advantages of
Albino Rats are: docile and small in size, easy to handle, a drug to be tested is
Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 121-142 121
CHAPTER 7
Product Development of Herbal Medicine
Ahmed Nawaz Khan1,*, Chandra Kala2 and Javed Ahmad3
1 School of Pharmacy, Graphic Era Hill University, Dehradun. Uttarakhand, 248002, India
2 Faculty of Pharmacy, Maulana Azad University, Jodhpur, Rajasthan, 342802, India
3 Department of Pharmaceutics, College of Pharmacy, Najran University, KSA
Abstract: Treating diseases with medicinal plants is considered as the oldest
therapeutic method that provides relief from illness to the whole humankind. In
developing countries, a large group of the population have been found using
phytotherapy for centuries, and still, they are following the same remedies. Preparation
and compounding of one or more herbs containing phytochemicals lead to the
production of finished herbal products, which are formulated from dried roots and
extracts. Widespread use of these products requires quality and safety with good
manufacturing practices and stringent evaluation criteria. From basic to advance, this
chapter addresses the challenges and procedures ranging from harvesting, selection of
plants, extraction, and formulation to the product approval by following the good
agriculture, laboratory practice and good clinical practice and it would be a helpful
medium for consideration during the product development period. As a whole, this
chapter gives an overview to academicians, researchers, and industrial personnel for a
better understanding of herbal products.
1. INTRODUCTION
Being healthy is the right of every human, and this right is a state of complete
emotional and physical wellbeing that is necessary for living a happy life [1].
Additionally, when health gets disrupted by any weaknesses; nature serves with
multiple interventions by its phytochemicals (in Greek phyto means plant). These
phytochemicals are important for the growth, development, and protection of
plants; and they also help in the cure of many human and veterinary diseases [2].
With good trust, these naturally occurring chemical moieties in plants are also
responsible for the color, odor, and flavor; and are considered to be a major cont-
* Corresponding author Ahmed Nawaz Khan: School of Pharmacy, Graphic Era Hill University, Dehradun.
Uttarakhand, India 248002; E-mail: ahmednawaz4u@gmail.com
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
Keywords: Dosage Form, Good Manufacturing Practice, Herbal Products, Herbal
Preparations, Phytochemicals, Phytoconstituents.
122 Bioactive Phytochemicals: Drug Discovery to Product Development Khan et al.
ributor for protecting the health [3]. Phytochemical products are obtained from
dried roots (herbal material) and extracts (herbal preparation) of the plant or herbs
[4]. These plants contribute 11% of the drugs listed in 252 drugs of the World
Health Organization's (WHO) essential medicines [5]. Surprisingly, there are
about 121 bioactive phytochemicals commonly prescribed by the medical
practitioners, which are among one-fourth of the drugs prescribed across the globe
[6].
Treating diseases with medicinal plants is considered as the oldest therapy system
that provides relief from illnesses to the humankind. In India, such systems are
extensively followed through Ayurvedic, Unani, and Siddha system of medicine;
and now Aamchi which also refers to as Tibetan Medicine System has also been
included. Globally, these systems come under complementary medicines which
are also referred to as integrating, naturopathy, or alternative medicine system [7].
Being therapeutically active, medicinal plants are used from Ayurveda to Chinese
traditional medicines, Unani to Aamchi and Amazonian to African medicines,
etc., which integrate phytotherapy in these systems irrespective of their versatile
theoretical and cultural values [8]. Such alternatives may provide an authentic and
efficacious therapeutic treatment in case of many incurable diseases; and can help
practitioners to accept a new and interesting line of treatment. Along with the
right to choose treatment methods or therapies; in general, people are free to
select bioactive phytochemicals products or phytomedicines on a self-selection
basis. Day by day, demand for herbal products is increasing due to growing
consumers’ interest in alternative or natural therapies [9]. More than three fourth
of the Indian population prefer phytochemical products for treatment, [10]
moreover, up to 90% of Africans depend on traditional medicines and surprisingly
traditional medicines in China account for 40% of the health delivery system [11].
However, with a new framework of evidence-based medicines; the development
of these traditional products becomes a challenge for pharmaceutical industries.
Under the correct scientific and ethical standards for research and development,
there are big challenges like lack of scientific validation and standardization, lack
of quality and regulatory aspects, limited evidence-based studies on efficacy and
safety, and lack of pharmacokinetic studies of bioactive compounds Thus
scientific validity of herbal products is still often questioned. Besides these
challenges and to understand the herbal product development, this chapter
highlights current issues and gives insights into harvesting and regulatory
procedures, including clinical and nonclinical aspects, for the development of
bioactive herbal products.
Product Development of Herbal Bioactive Phytochemicals: Drug Discovery to Product Development 123
1.1. Scope of Herbal Medicinal Products
Consumption of phytochemical products declined when synthetic medicines
evolved; however, for the past few decades, phytotherapy is again booming in all
industrialized countries. Interestingly in developing countries, a large group of the
population has been found to use phytotherapy for centuries, and still, they are
following the same remedies. It has been observed that when synthetic medicines
proved ineffective in chronic diseases like cancer or new infectious diseases, the
demand for these herbal products increased [12]. Furthermore, continuing
research on ethnobotanics is leading to the discovery of new phytochemicals
present in nature for the treatment of various major pathologies and are opening
new ways for drug development [13]. Due to affordability, fewer side effects, and
easy accessibility even without prescription make these phytochemical products
highly acceptable by the consumer.
2. CHALLENGES IN HERBAL MEDICINAL PRODUCT
DEVELOPMENT
Phytochemical products have reached extensive acceptability across the globe as
therapeutic and nutritional products. However, quantitative and qualitative
analysis of the phytochemical composition of bioactive compounds or markers is
a major challenge. Science admits that consuming food rich in these chemical
compounds has health benefits however, pieces of evidence exist to prove the
specific recommendation for these phytochemical intakes. Even there are some
known health effects of phytochemicals that researchers have not recognized yet.
Moreover, it is known that 'Herbal product is free of side effects', but this is not
completely true [14]; as many extensive scientific studies reject this myth.
Clinical data reveals the real picture like cardiovascular ailments with the use of
ephedra, hepatotoxicity by kava-kava consumption, and water retention by
licorice [15, 16]. Many other studies have reported numbers of side effects and
adverse reactions in consumers for a wide range of mechanisms [9]. In a WHO
database, 4 million reports from 100 countries across the globe are indicated out
of which 21,000 discussed the adverse effects of natural products [17]. Another
study from American Poison Centres revealed that a user consuming supplements,
herbal and homeopathy products fell into a category which was linked to hospital
admission [18, 19]. These Adverse Drug Reactions (ADR) negatively affect the
promotion of herbal products, and the most common reason of these adverse drug
reactions is attributed to the presence of adulterants, undeclared substances,
synthetic chemicals, or some other active ingredients which are common during
product development or trading of raw medicinal plants [6]. The adulteration
sometimes is unintended and may be due to the case of substitution between
Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 143-157 143
CHAPTER 8
Quality Control of Herbal Medicinal Products
Javed Ahamad1,*, Esra T. Anwer2, Muath Sh. Mohammed Ameen2, Jamia
Firdous3 and Nehal Mohsin4
1 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Kurdistan
Region, Iraq
2 Department of Pharmaceutics, Faculty of Pharmacy, Tishk International University, Kurdistan
Region, Iraq
3 Department of Pharmacy, Institute of Bio-Medical Education and Research, Mangalayatan
University, Aligarh, India
4 Department of Clinical Pharmacy, College of Pharmacy, Najran University, Kingdom of Saudi
Arabia
Abstract: The quality control of medicinal agents derived from natural sources is of
paramount in ensuring safety and efficacy. The major hindrance in the acceptance of
herbal medicines into modern medical practices is the lack of scientific and clinical
data on the safety and efficacy of the herbal products. In general, there is a lack of strict
guideline for quality control of herbal medicinal products used for the treatment of
various human diseases. In recent years, due to enormous increased interest in herbal
medicine, we need strict quality control parameters for safe and efficacious herbal
medicine. Several quality control parameters (e.g. organoleptic, morphological,
physico-chemical, chromatographic and toxic substances) are mentioned in
Pharmacopoeias like Indian Pharmacopoeia (IP), British Herbal Pharmacopoeia (BHP),
Ayurvedic Pharmacopoeia of India (API) and WHO guidelines etc. Chromatographic
techniques, such as TLC, HPTLC, HPLC, GC-MS; and toxic substances such as
aflatoxins, heavy metals, pesticide residues, microbial load determinations, are
important parameters considered for quality control of herbal drugs. This book chapter
provides a critical overview of different quality control parameters of herbal drug,
which are necessary for compliance of regulatory guidelines of several developing and
developed countries.
* Corresponding author Javed Ahamad: Department of Pharmacognosy, Faculty of Pharmacy, Tishk International
University, Kurdistan Region, Iraq; E-mails: jas.hamdard@gmail.com, javed.ahamad@tiu.edu.iq
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
Keywords: HPTLC, Herbal Products, Pharmacopoeia, Quality Control,
Standardization, WHO Guidelines.
144 Bioactive Phytochemicals: Drug Discovery to Product Development Ahamad et al.
1. INTRODUCTION
Standardization means adjusting the preparation of herbal drugs to a defined
content of a constituent or group of substances having known therapeutic activity.
Medicinal plants are widely used in herbal based drug formulations. While herbal
medicinal products are often viewed as natural and healthy, they are not free from
adverse effects. Such adverse effects of herbal drugs can occur due to
adulteration, substitution, contamination, incorrect preparation and the most
significant lack of standardization [1, 2]. The main criticism confronting the
traditional medicine system is inadequate scientific validation and standards of
plant material used by the manufacturer in herbal formulations. Quality control of
herbal medicines is of paramount importance for its adoption in modern medical
practices. The major problem with herbal medicine acceptance is the lack of data
on herbal formulation on safety and efficacy. Standardization or quality control is
of paramount importance to ensuring the therapeutic activity and safety of herbal
drugs [3]. There are no standards for quality control of medicinal plants that are
used in herbal formulation in most underdeveloped and developing countries.
Quality control of herbal formulations is more complicated than synthetic
formulations because of the complex nature of herbal drugs comprising several
phytochemical compounds [4].
Usually, one or two most important chemical compounds chosen for analysis are
used for quantitative analysis of bioactive phytochemicals in herbal formulations.
Nonetheless, this type of determination does not give a complete picture of the
formulation, as multiple constituents are usually responsible for their therapeutic
effects [5]. Several chromatographic techniques, such as HPTLC, HPLC, GC,
GC-MS, and LC-MS can be applied for this kind of documentation for quality
control of herbal formulations and individual medicinal plants. For the purposes
of chemical standardization and quality control of individual herbs and herbal
formulations, chemical fingerprints obtained by chromatographic techniques are
strongly recommended. Therefore, there is a strong need to promote
standardization of quality parameters and formulations of important medicinal
plants. WHO has emphasized the need to ensure quality control of medicinal plant
products through the use of modern techniques and the application of appropriate
standards. Several pharmacopoeias, including the United States Pharmacopoeia
(USP), Indian Pharmacopoeia (IP), British Pharmacopoeia (BP), and British
Herbal Pharmacopoeia (BHP), cover monographs and quality control tests for a
few of the medicinal plants used in their respective countries [6]. This chapter of
the book provides an overview of the various quality criteria applied to
standardize herbal medicines.
Herbal Medicinal Products Bioactive Phytochemicals: Drug Discovery to Product Development 145
2. THE NEED OF STANDARDIZATION
In general, adverse effects are thought to be less with herbal drugs compared to
modern drugs, but reports on serious side effects indicate the need to develop
effective marker-based standardization of herbal formulations [7]. Because of the
lack of quality control measures on herbal formulations, people cannot make use
of the benefits of these medicines. Because of scientific awareness, a scenario has
been created for conducting research activities such as standardizing herbal
formulations and developing scientific methods for large-scale production [8].
The idea of standardization is to establish consistent therapeutic effectiveness of
batch-to-batch natural products [9 - 11].
3. METHODS OF STANDARDIZATION OF HERBAL MEDICINAL
PRODUCTS
In recent years, due to enormous increased interest in herbal medicine, we need
strict quality control parameters for safe and efficacious herbal medicine. Several
quality control parameters such as organoleptic, morphological, physico-
chemical, chromatographic (e.g. TLC, HPTLC, HPLC, GC-MS) and toxic
substances (e.g. aflatoxins, heavy metals, pesticide residues, microbial load
determinations) are mentioned in pharmacopoeias and WHO guidelines for
ensuring safety and efficacy of herbal medicinal agents obtained from natural
products [1, 12].
3.1. Botanical Parameters
3.1.1. Morphological Characters
Detailed study of morphological character may be useful in identifying various
species of plants. A drug's morphological study includes the naked-eye visual
appearance. Macroscopic analysis explores the following characters of the plant:
size, shape, colour, taste, odour, fracture, etc. [6].
3.1.2. Powder Microscopy
Very small amounts of powdered drugs are taken on a slide and treated with
gentle heating with HCl and chloral hydrate, and stained with phloroglucinol.
After mounting with glycerin, observed under a compound microscope at suitable
magnification. Powder microscopy helps in determining different types of tissues
and cells present in plant materials such as phloem fibers, xylem vessels,
158 Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 158-175
CHAPTER 9
Regulatory perspectives of Herbal Medicinal
Products
Faraat Ali1,*, Shaik Khasimbi2, Kamna Sharma3, Manisha Trivedi4, Asad Ali5
and Javed Ahmad6,*
1 Laboratory Services, Botswana Medicines Regulatory Authority, Plot 112 International Finance
Park, Gaborone, Botswana
2 Department of Pharmaceutical Chemistry, Delhi Institute of Pharmaceutical Sciences and
Research (DIPSAR), Mehrauli-Badarpur Road, Push Vihar, Sector-3, New Delhi, 110017, India
3 Department of Pharmaceutical Analysis, Indo-Soviet Friendship College of Pharmacy, Moga,
Punjab, India
4 NIMS University, Jaipur, Rajasthan, India
5 Department of Chemistry, School of Chemical and Life Sciences, Jamia Hamdard University,
New Delhi, India
6 Department of Pharmaceutics, College of Pharmacy, Najran University, KSA
Abstract: Quality control of medicinal agents derived from natural sources is
paramount in ensuring safety and efficacy. In the modern medical practice, the major
obstacles for the acceptance of herbal medicines are the lack of scientific and clinical
data on safety, quality and efficacy in herbal products. In general, there is no quality
control of medicinal products in many countries. Strict regulatory guidelines on herbal
products are followed only in few countries like Canada, Europe, Australia, the USA,
and Japan. In recent years, due to the enormous increased interest in herbal medicines,
we need strict regulatory guidelines to ensure safety and efficacy of herbal medicine.
The regulatory guidelines should focus mainly on laws related to the registration of
herbal medicines for manufacturing and marketing. This book chapter will provide a
comprehensive overview of a regulatory guidelines, required for manufacturing and
marketing of herbal medicines in regulated, semi-regulated and unregulated markets.
This book chapter also comprises discussion over global need of harmonization of
regulatory guidelines to ensure safe and effective use of herbal medicines.
Keywords: Efficacy, Herbal Medicines, Harmonization, Pharmacopoeia,
Regulatory Agency, Safety.
* Corresponding author Faraat Ali and Javed Ahmad: Laboratory Services, Botswana Medicines Regulatory
Authority (BoMRA), Gaborone, Botswana and Department of Pharmaceutics, College of Pharmacy, Najran University,
KSA; E-mails: FRHTL6@gmail.com & jahmad18@gmail.com
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
Herbal Medicinal Products Bioactive Phytochemicals: Drug Discovery to Product Development 159
1. INTRODUCTION
Generally, it is to be believed that the risks associated with herbal drugs are
minimal, but reports on serious adverse effects show the need for strict regulatory
guidelines. The drug discovery on the basis of the natural product has experienced
indefinite number of challenges, such as identification, availability of bioactive
ingredients, etc. Although diverse approach including novel and potential
therapeutic agents modulating cell signaling cascades are popular, for the drug
discovery of natural products from medicinal plants [1]. The drug discovery of
medicinal plants have played an important role in curing cancer (e.g. Vincristine,
Vinblastine, Paclitaxel, Docetaxel, etc.) and malaria (e.g. Quinine, Artemisinin).
The drug discovery from plants today is an expensive and time-consuming
process, it requires a team efforts consisting of experts from different disciplines,
such as pharmacognosists, pharmacologists, medicinal chemist and
pharmaceutists. The trend today, especially in an industrial setting, is to seek
bioactive phytochemicals that will serve as a lead compound in drug discovery
and used for synthetic or semi-synthetic drug development, to make sure patent
protection [2]. Newman and Cragg, reported that totally 1562 new drugs approved
during 1981 to 2014, out of which natural products (04%; N), natural product
derivatives (21%; ND), synthetic drugs with natural-derived pharmacophores
(10%; S*/NM), and synthetic drugs designed based on knowledge gained from a
natural product (11%; S/NM). The above reports suggested that the natural
products are important source, hence involved in the new drug development as
natural drugs or semi-synthetic drugs [3]. The plants considered as equal or
superior in the drug discovery because of their chemical diversity and human
friendly nature based on their long history of use for consumption.
The natural medicinal product has played an important role to give basic
healthcare requirements of the population. The World Health Organization
(WHO) has defined natural medicine along with allopathic medicines for
improvement and prevention of illness. In Asian countries such as Korea, China,
India and Japan, the natural medicinal plants plays a vital role in curing different
diseases. However, each country has its own unique traditional systems such as
the traditional Indian medicine (TIM), traditional Korean medicine (TKM),
traditional Chinese medicine (TCM) and traditional Japanese medicine (TJM). In
India, there are seven well-known different systems of medicines i.e. Ayurveda,
Yoga, Siddha System of Medicine (SSM), Unani Medical System (UMS),
Naturopathy, Homoeopathy, and Amchi Medical System (AMS) [4 - 7]. The plant
materials used in many herbal medicinal products manufactured for general retail
sale are not standardized. The quality control system in the development for
various herbal preparations is much more difficult than for synthetic drugs
because of the chemical complexity of their ingredients. As herbal preparations
160 Bioactive Phytochemicals: Drug Discovery to Product Development Ali et al.
are composed of 100 unique or species-specific compounds, various difficulties
are completely characterized by all these compounds. Thus, there is a pertaining
need in promoting standardization for various quality parameters in medicinal
plants and formulations. WHO emphasized the need to ensure quality control of
herbal plant products by using modern techniques and applying suitable standards
[8]. Several pharmacopoeias including United States Pharmacopoeia (USP),
Indian Pharmacopoeia (IP), British Pharmacopoeia (BP), and British Herbal
Pharmacopoeia (BHP) covers monographs and quality control tests for few of the
medicinal plants used in the respective countries (Table 1).
Table 1. Regional Regulating Bodies in different countries.
Countries Regulatory Bodies
India Ayurveda, Yoga and Naturopathy, Unani, Siddha and Homoeopathy (AYUSH)
Australia Therapeutic Goods Administration (TGA)
Canada Health Canada
China State Food and Drug Administration
Europe European Medicine Agency (EMA)
Malaysia National Pharmaceutical Control Bureau
USA United States Food and Drug Administration (USFDA)
Singapore Health Science Authority (HSA)
South Korea Korean Food and Drug Administration (KFDA)
Saudi Arabia Saudi Food and Drug Authority (SFDA)
Japan Pharmaceutical and Medical devices Agency (PMDA)
Indonesia National Agency of Drug and food control (NADFC)
Brazil Agencia Nacional de Vigilancia Sanitaria (ANVISA)
South Africa South African Health Products Regulatory Agency (SAHPRA)
New Zealand New Zealand Medicine and Medical Devices Safety Authority
United Kingdom Medicines and Healthcare Products Regulatory agency ((MHRA)
Tanzania Tanzania Food and Drug Administration (TFDA)
Botswana Botswana Medicine Regulatory Authority (BoMRA)
In the recent years traditional medicines has been regulated by different
authorities of the respective countries to ensure the safety and efficacy of these
drugs [9]. Various regulatory authorities have designed the guidance for the
effective and safe use of medicines to avoid the adverse effect and to improve
healthcare. WHO has proposed various guidelines for the herbal products that act
as a reference standard for various countries to keep up the quality and safety of
176 Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 176-213
CHAPTER 10
Phytochemicals for the Treatment of Human
Diseases
1 Department of Pharmacognosy, Faculty of Pharmacy, Tishk International University, Erbil,
Kurdistan Region, Iraq
2 Department of Pharmacology, Faculty of Pharmacy, Tishk International University, Kurdistan
Region, Iraq
3 Department of Pharmacognosy & Phytochemistry, Faculty of Pharmaceutical Sciences, Rama
University, Rama City, Mandhana, Kanpur (Uttar Pradesh) - 209 217, India
4 Department of Pharmaceutics, Faculty of Pharmacy, Tishk International University, KRG, Iraq
5 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Tishk International University,
KRG, Iraq
6 Department of Clinical Pharmacy, College of Pharmacy, Najran University, Kingdom of Saudi
Arabia
Abstract: Medicinal plants are a major source of remedies for the treatment of human
ailments in under-developed and developing countries. Traditional or alternative
medicines were practiced in ancient civilizations for the cure of human ailments. In
recent years, natural products play a vital role in drug discovery for life-threatening
ailments like cancer, malaria, diabetes, and cardiovascular problems. Due to the
advancement of scientific techniques, isolated phytochemicals can be developed as a
medicine for lifestyle and chronic disorders. Herbal medicinal products are also
developed for effective treatment of several diseases like cancer, malaria, diabetes,
cardiovascular complications, etc. Recently,drug discovery from plants for the
treatment of cancer has become more focused, leading to the discovery of novel
anticancer drugs such as paclitaxel, docetaxel, topotecan, irinotecan, vincristine, and
vinblastine, etc. In this book chapter, we have discussed important medicinal plants and
bioactive natural products for the treatment or management of diabetes mellitus,
cancer, obesity, and cardiovascular complications.
Keywords: Bioactive Phytochemicals, Cancer, Cardiovascular Complications,
Drug Discovery, Diabetes Mellitus, Herbal Medicine, Natural Product.
* Corresponding author Javed Ahamad: Department of Pharmacognosy, Faculty of Pharmacy, Tishk International
University, Erbil, Kurdistan Region, Iraq; E-mails: jas.hamdard@gmail.com; javed.ahamad@tiu.edu.iq
Javed Ahmad and Javed Ahamad (Ed.)
All rights reserved-© 2020 Bentham Science Publishers
Javed Ahamad1,*, Subasini Uthirapathy2, Kamran Javed Naquvi3, Muath Sh.
Mohammed Ameen4, Esra T. Anwer4, Abdul Samad5 and Mohammad
Shabib Akhtar6
Treatment of Human Diseases Bioactive Phytochemicals: Drug Discovery to Product Development 177
1. INTRODUCTION
Higher plants fulfill the significant human requirements such as food, shelter,
clothes, and remedies for the cure of ailments [1]. Ethnobotany and
ethnopharmacology utilize plants for the treatment of various ailments based upon
the long history of their use by humans. Natural products derived from medicinal
plants considered safe and effective compared to modern synthetic drugs. Because
these natural resources are distributed throughout the world and grown in crop
fields, they are comparatively cheap to synthetic drugs. The WHO estimates about
traditional medicines states that approximately 80% of developing and developed
countries rely on herbal or traditional medicine for their primary health care needs
[2, 3].The plant has advantages in drug discovery based on along history of use by
humans as food and medicine. So ,the drugs discovered from medicinal plants are
supposed to have fewer side effects compared to modern drugs. In recent years,the
discovery of several novel and potent anticancer (e.g. taxols, vincristine,
vinblastine, camptothecin etc.), antimalarials (e.g. artemisinins, quinine etc.),
antihypertensive (e.g. reserpine), cardiotonics (e.g. digoxin) etc realizes the
importance of natural resources as the major source of drug discovery [3]. Hence,
public, academic, and government interest is increasing nowadays in traditional
medicine [4]. Alternative or traditional medicines use plants and have become a
significant source for the treatment of chronic diseases and disorders of humans
[5]. Current therapy to alleviate cancer and metabolic disorders such as diabetes
mellitus, cardiovascular complications is not optimal, and thus, efforts have been
made to develop effective and better drugs from natural sources [6, 7]. Isolated
bioactive phytochemicals and their derivatives also approved for the treatment of
human ailments' such as malaria, cancer, diabetes, etc. (Table 1). In this book
chapter, we discussed separately natural products used in the treatment of diabetes
mellitus, cancer, cardiovascular complications, obesity, and natural products with
anti-inflammatory and antioxidant activities.
Diabetes mellitus (DM) is a chronic metabolic disorder, associated with obesity
and cardiovascular complications, which emerged as the major killer for mankind
[8]. Sedentary lifestyle and obesity nowadays have become a major factor in the
development of type 2 DM, and diabetes is the primary cause of other severe
health problems such as diabetic retinopathy, neuropathy and cardiovascular
complications such as atherosclerosis, stroke, peripheral vascular diseases,
coronary artery diseases and these lead to angina and diabetic myonecrosis [9]. In
recent years, it has been observed that the number of DM patients worldwide
increased significantly. Current therapies for the treatment of type 2 DM and
2. HERBAL MEDICINES WITH ANTIDIABETIC ACTIVITY
178 Bioactive Phytochemicals: Drug Discovery to Product Development Ahamad et al.
related cardiovascular complications are not optimal and having more adverse
effects, and thus efforts should be made to develop effective and safe drugs from
natural sources. In recent years, the use of traditional medicines in the modern
way has expanded globally, and it's gaining popularity day by day. Medicinal
plants have been widely used for therapeutic purposes due to the expensive and
unaffordability of modern medicines in the rural areas of developing and
undeveloped countries compared to herbal remedies, which are cheap and
considered as safe [10 - 12]. In pursuit of new medications for diabetes mellitus
treatment and associated complications, several researchers emphasize the
utilization of natural products and their secondary metabolites. Ethnobotanical
knowledge reports about 800 plants that possess the antidiabetic potential [13].
Table 1. Natural products (N) and natural products derivative (ND) approved by US-FDA for the
treatment of human diseases [7].
Clinical Class Natural Products/ Natural Product Derivatives Year of Approval
Antibacterial drugs Carumonam (N) 1988
Daptomycin (N) 2003
Fidaxomicin (N) 2011
Fosfomycin trometamol (N) 1988
Isepamicin (N) 1988
Micronomicin sulphate (N) 1982
Miokamycin (N) 1985
Mupirocin (N) 1985
Netilimicin sulphate (N) 1981
RV-11(N) 1989
Teicoplanin (N) 1988
Antiparasitic drugs Artemisinin (N) 1987
Arteether (ND) 2000
Artemether (ND) 1987
Artesunate (ND) 1987
Mefloquine HCI (ND) 1985
214 Bioactive Phytochemicals: Drug Discovery to Product Development, 2020, 214-221
Javed Ahmad and Javed Ahamad (Eds.)
All rights reserved-© 2020 Bentham Science Publishers
SUBJECT INDEX
A
Abbreviated new drug application (ANDAs)
164
ACC synthase 22
Acetic acid 21, 77, 108, 150
glacial 77
Acid 39, 40, 41, 42, 193, 194, 197, 198, 200
ascorbic 197
betulic 197
betulinic 193, 194
caffeic 40, 198
carnosic 39, 41, 200
chlorogenic 40, 41, 42
glycyrrhetinic 198
protocatechuic 193
rosameric 198
rosemarinic 198
rosmarinic 39, 41
Activity 9, 15, 23, 28, 150, 185, 198, 200
antiobesity 198, 200
biochemical 23
fluorescence 150
pharmacologic 9
Adverse Drug Reactions (ADR) 123
Agents 28, 55, 74, 109, 110, 111, 128, 129,
130, 137, 183, 192, 196
anti-inflammatory 109, 192
buffering 129
chelating 137
coating 128
dehydrating 28
emulsifying 130
hypnotic 110
hypocholesterolemic 183
phlogistic 109
synthetic 192
therapeutic 55, 74
Alcohol 46, 112
absolute 112
monoterpene 46
Aliphatic 90, 91
acid 90, 91
chain 91
nature 90
Alkaline sodium picrate 77
Alkaloids 14, 32, 37, 40 43, 44, 47, 74, 77, 84,
150, 186, 188, 189, 197, 198
indole 47, 186
purine 40, 44
quinoline 188
Alkaloids nitrogenous compounds 23
Amchi medical system (AMS) 159
AMPK activation 182
Amyloglucosidase 40
Analgesic activity 107, 108
Analgesics 6, 101, 104, 107, 108, 192
acting 107
moderate 104
Analysis 7, 31, 49, 87, 123, 136, 144, 153,
154
animal safety 7
qualitative 123, 153
qualitative protein 87
quantitative 31, 49, 136, 144, 153, 154
Anti-atherosclerosis activity 114
Anti-atherosclerotic property 196
Anti-diabetic Activity 115
Antihistamine medications 115
Antihyperglycemic activity and antidiabetic
effect 185
Antioxidant 114, 177, 190, 192, 193, 194
activity 177, 190, 192, 193, 194
enzymes 114
Artemisinin-based combination therapy 46
Asiatic acid 190
Assay 10, 115
α-glucosidase enzyme inhibition 115
systemic pharmacological 10
Atherogenesis 114
Atherosclerosis 115, 177, 191, 195, 196, 197
effective 115
Atomic absorption spectroscopy (AAS) 152
Ayurvedic pharmacopoeia committee (APC)
168
Subject Index Bioactive Phytochemicals: Drug Discovery to Product Development 215
B
Balance of natural soil fertility 23
Bioactive phytochemical(s) 42, 122, 133, 137
isolation 42
products 122, 133, 137
Blood 101, 115, 185
collection techniques 101
glucose levels 115
fasting 185
Boswellic acid 190
Botanical drug products 132, 167
Breast 190, 191
berberis vulgaris 190
panaxatriol Panax ginseng 191
solamargine Solanum nigrum 191
British herbal pharmacopoeia (BHP) 143, 144,
160, 163
C
Cancer 1, 2, 7, 123, 151, 176, 177, 186, 187,
188, 190, 191, 196
allicin Allium sativum Colon 190
cell lung 188
colon 191
lung 191
neuroectodermal 190
prostate 191
skin 190
testicular 187
testis 191
thyroid 191
Cardiovascular 54, 194, 196
diseases 54, 196
disorders 194
Cellulase 40, 41
Center for drug evaluation and research
(CDER) 166, 167
Centrifugal partition chromatography (CPC)
57
Chemical 19, 144
fertilizers nitrogen 19
fingerprints 144
Chemical structure of 43, 44, 47, 48, 89, 90,
91, 180, 181, 183, 184, 187, 189, 190
4-Hydroxyisoleucine 184
artemisinin 47
berberine 183
caffeine 44
camptothecin 189
charantin 180
colchicine 189
digitoxigenin 48
epicatechin 184
erythrocentaurin 89
etoposide 187
norgadosic acid 91
octatriacontenoic acid 90
quinine 43
reserpine 47
solasodine 44
swertiamarin 181
trans-resveratrol 190
Vincristine 187
Chinese 163, 170, 171
herbal medicines (CHM) 170
pharmacopoeia (ChP) 163, 170, 171
Chromatographic techniques 65, 134, 143,
144, 154
Chromatography techniques 65
Collision-induced dissociation 87
Column Chromatography (CC) 47, 49, 54, 56,
59, 60, 62, 66, 89, 91, 92
adsorption 59
principles 62
Concentration 14, 15, 23, 33, 34, 48, 76, 107,
131, 136, 151
intra-cellular 48
secondary metabolite 15
Contamination 25, 27, 133, 136, 144, 152, 191
microbial 136
microorganism 25
Conversation agriculture (CA) 15
Countercurrent chromatography 57
droplet 57
dual flow 57
Crude caffeine 45
Cultural food habits 195
Curative plants 17, 18, 20, 24, 25, 28
harvested 25
protecting 20
stuffs 24, 25, 28
D
Death 106, 107, 115, 186, 194
human 186
Decoction 36, 37, 38
216 Bioactive Phytochemicals: Drug Discovery to Product Development Ahmad & Ahamad
solvent High Economical 38
extraction methods 37
method 36, 37
De novo synthesis 23
Diabetes 1, 2, 7, 115, 176, 177, 180, 181, 182,
184, 195
induction of 115
treatment of 180, 181, 182, 184
Diabetes mellitus (DM) 115, 176, 177, 178,
179, 180, 182, 198
treatment of 177, 178, 179, 180
Diabetic 177
myonecrosis 177
retinopathy 177
Dietary supplement health education act
(DSHEA) 166
Diseases 1, 7, 8, 10, 55, 100, 103, 117, 121,
122, 127, 133, 177, 186, 187, 191, 192,
193, 195, 196
autoimmune 192
coronary artery 177, 195
cure of 1, 8
deadly 186
gastrointestinal 195
heart 195
immunological 103
painful 193
peripheral vascular 177
skin 187
vascular 196
veterinary 121
Disorders 103, 133, 177
chronic metabolic 177
endocrine 103
DNA bar-coding system 170
Droplet counter-current chromatography
(DCCC) 58, 74
Drug 114, 115, 136, 168
formulations 114, 168
-herb interaction 136
-induced hyperglycemia 115
Drug material, avoided confusion medicinal
plant 27
Drug development 55, 66
synthetic 55
process 66
Drug discovery 55, 190
cascade 190
process 55
Dyslipidemia 196
E
Effectiveness 131, 145, 161
consistent therapeutic 145
Effects 101, 102, 111, 114, 134, 136, 144,
180, 184, 192, 196, 200
anti-atherogenic 114
anti-diabetic 184
antihyperlipidemic 180
anti-inflammatory 192
anti-obesity 200
anti-platelet 196
anxiolytic 111
hypolipidemic 196
strong plaque stabilizing 196
therapeutic 102, 134, 136, 144
toxicodynamic 101
Electrolytes 114
Electromagnetic radiation 76
Enzyme assisted extraction (EAE) 31, 38, 40,
42, 49
Enzymes 23, 26, 31, 38, 40, 41, 42, 49, 114,
181, 192
carbohydrate metabolizing 181
cellulase 41
cell wall dissolving 23
lipometabolizing 114
respiratory 114
Ethno-pharmacological approach 8
European medicines agency 131, 160, 165,
171
Evaluation of anti-atherosclerosis activity 114
Evaluation of 33, 34, 35, 36, 37, 38, 39, 40,
109, 112, 113, 114, 115
anti-atherosclerosisactivity of herbal
medicine 114
anti-depressant effect of herbal medicine
113
anti-diabetic activity of herbal medicine
115
anti-inflammatory activity of herbal
medicine 109
anti-peptic-ulcer activity of herbal
medicine 112
bioactive 38, 39
medicinal plants 33, 37
phytochemicals 34, 35, 40
plant materials 33, 35
thermolabile drugs 36, 37
thermolabile herbal drugs 35, 39
Subject Index Bioactive Phytochemicals: Drug Discovery to Product Development 217
F
Fingerprint 80
Finger printing 153
Flash chromatography (FC) 32, 54, 62
Fluorescence analysis 150
Fractionation 7, 62, 127, 131
bioassay-guided 7
FT-IR and NMR spectroscopy 73
G
Gallic acid 59, 184, 193, 198
Galloylquinic acids 64
Gas chromatography 151, 153
-mass spectrometry 153
Gibbrelic acid 21
Glucose 40, 41, 185
metabolism 185
oxidase 40, 41
Glucose consumption 182
enhanced 182
Glutathione 114
peroxidase 114
reductase 114
Glycyrrhizinic acid 76
Good 121, 126, 131, 132, 138, 139, 164
agricultural and collection practices
(GACP) 126, 131
clinical practice (GCP) 121, 126
herbal processing practice (GHPP) 126,
131
laboratory practice (GLP) 126
manufacturing practice (GMP) 121, 126,
131, 132, 138, 139, 164
Gymnemic acids 39, 41, 181, 182
arylated 181
H
Healthcare 160, 170
products regulatory 160
system 170
Health 152, 160
hazards 152
science authority (HSA) 160
Heavy metal toxicity 100, 152
Hemicellulose 40
Hepatic lipase 114
Hepatocarcinoma 191
Herbal drugs 31, 32, 124, 125, 126, 127, 131,
143, 144, 145, 146, 151, 152, 165, 166
crude 151
powdered 131
Herbal material 122, 124, 125, 127, 128, 129,
130, 131, 133, 138
boiling 129
dried 128
Herbal medicinal 117, 123, 131, 134, 145, 167
agents 145
and molecular study 117
compendium (HMC) 167
ingredients 167
product consistency 134
product development 123, 131
Herbal products medicinal committee
(HPMC) 165
Heteronuclear 85, 86
multiple-bond coherence 86
single quantum coherence 85
Hetrocyclic compounds 23
High-performance liquid chromatography
(HPLC) 133, 134, 143, 144, 145, 153,
154
High performance thin layer chromatography
(HPTLC) 143, 144, 145, 151, 153, 154,
155
High Pressure Liquid Chromatography 153
High throughput screening (HTS) 55
Hodgkin’s 190, 191
disease 191
lymphoma 190
Homoeopathic systems 167
Hydrochloric acid 146, 147, 150
Hypnosis 110
Hypnotic effect of herbal medicine 110
I
Immune system responses 192
Indole acetic acid 21
Insect growth regulator (IGR) 20
Insulin 179
exogenous 179
Insulinomimetic activities 185
Insulin resistance 180, 185, 186
218 Bioactive Phytochemicals: Drug Discovery to Product Development Ahmad & Ahamad
K
Korean food and drug administration (KFDA)
160
L
Liquid 22, 40, 45, 59, 127, 130, 134, 191
immiscible 130
supernatant 45
Liquid chromatography (LC) 56, 59, 62, 63,
73, 74, 86, 88, 153, 154
medium pressure 56, 59, 62, 63, 74
preparative high-performance 73, 74
preparative high pressured 56
-mass spectrometry 154
M
Maceration technique 36
Macroscopic analysis 145
MALDI-TOF Mass Spectrometry 87
Manglanostenoic acid 60
Mass spectrometry (MS) 73, 74, 75, 86, 96,
133, 154
techniques 86
Mass spectroscopy 86, 133, 153, 154
gas chromatography 151
Medicinal materials 170
database 170
DNA Barcode Database 170
DNA barcode database (MMDBD) 170
Medicinal plants 15, 25, 54, 159, 189
cultivation of 15, 189
economic 25
natural 159
time immemorial 54
Medicines 15, 26, 46, 101, 122, 123, 159, 166,
167, 170, 186
allopathic 159
anticancer 186
complementary 122
cough 46
crude 15, 26
defined natural 159
derived 167
evidence-based 122
natural 166
plant-based 101
proprietary Chinese 170
synthetic 123
traditional Indian 159
traditional Japanese 159
traditional Korean 159
Medicines and healthcare products regulatory
160
Medium pressure liquid chromatography
(MPLC) 56, 59, 62, 63, 66, 74
Microorganisms 17, 19, 23, 25, 100, 135
aerobic 135
non-symbiotic 19
soil inhabiting 23
Microwave-assisted extraction (MAE) 34, 36,
38, 39, 41, 49
Mixture of 61, 64, 180
β-sitosterol glucoside 180
chloroform and methanol 61
methanol and acetonitrile 64
Modern methods of extraction 38
MPLC and preparative gas chromatography
56
Multiple reaction monitoring (MRM) 87, 88
Mycobacterium tuberculosis 192
N
Natural 22, 56, 100, 159, 170
capacity modifiers 100
-derived pharmacophores 159
growth inhibitors 22
medicinal products 159, 170
product isolation 56
Natural products 35, 37, 40, 41, 73, 74, 86, 96,
154
extraction of 35, 40, 41
heat-stable 37
structural assessment of 86, 154
structural elucidation of 73, 74, 96
NMR 73, 74, 75, 82, 83, 85, 96
spectroscopic technique 85
spectroscopy 73, 74, 82, 83, 85
spectroscopy and mass spectrometry 73,
74, 75, 96
Nonacosanoic acid 62
Non-steroidal anti-inflammatory drugs
(NSAIDs) 192
Norgadosic acid 91
Nuclear overhauser enhancement
spectroscopy 86
Subject Index Bioactive Phytochemicals: Drug Discovery to Product Development 219
O
Obesity 2, 176, 177, 196, 198, 199
controlling 196, 198, 199
management 196, 198
Octatriacontenoic acid 61, 90, 96
Organo chlorine pesticides (OCPs) 124
Origanum vulgare hydrodistillation method 37
P
Pest control 17, 19
Pesticides 19, 20, 124, 138, 151
synthetic 19
traditional 19
Pharmaceutical and medical devices agency
(PMDA) 160
Pharmacological 101, 152
screening techniques 101
monograph on herbal drugs 152
Phosphate-buffered saline (PBS) 113
Photochemical reference standards (PRS) 169
Physostigmine 6
Phytochemical 35, 40, 122, 123, 127, 128,
134, 137, 138
composition 123
constituents 128, 134
products 122, 123, 127, 134, 137, 138
breakdown of 35, 40
Plant materials 25, 26, 27, 33, 35, 127, 129,
144, 145, 148, 150, 151, 159, 163, 165
fragile medicinal 27
processed curative 26
raw medicinal 25
Plant metabolites, secondary 24, 149
Plant polysaccharides 137
Plants 1, 2, 4, 7, 8, 9, 10, 14, 15, 16, 17, 18,
22, 24, 26, 27, 28, 40, 55, 56, 74, 100,
121, 127, 131, 177, 186, 188
antidiabetic 185
cultivated 15, 28
fragmented 131
herbaceous 26
indigenous 9
medicinal drug 27
noxious 24
ornamental 186, 188
selection of 7, 8, 121
traditional 9
Preliminary phytochemical screening 75
Preparation 28, 180, 130, 131, 137
aromatic plant 28
exogenous insulin 180
herbal drug 131
oil-in-water 130
phytochemical 137
semisolid 130
Preparative 56, 65, 73, 74
gas chromatography 56, 65, 73, 74
thin layer chromatography 56
Pressure 40, 197
arterial blood 197
Pressurized liquid extraction (PLE) 31, 34, 38,
40, 41, 49
Products 24, 100, 122, 123, 124, 125, 126,
127, 128, 131, 132, 133, 134, 135, 136,
137, 138, 139, 144, 152, 160, 164
herb 24
herbal plant 160
homeopathy 123
medicinal plant 144, 152
nutritional 123
pharmaceutical 164
phytochemical combination 138
plant-based 100
traditional 122
Properties 2, 15, 24, 34, 40, 114, 189
agricultural 15
antipyretic 24
biological 15
exposed antidepressant 114
Protective index (PI) 112, 113
Proteins 87, 114, 130, 191, 193, 196
plasma 191
reactive 196
Protocatechnic acid 193
Proton NMR spectroscopy 83
Prunella vulgaris 191
Q
Quality control 131, 134, 143, 144, 145, 154,
169
measures 145
Methods 131
of herbal drugs 143, 154, 169
of herbal formulations 144
of herbal medicines 144
techniques, conventional 134
220 Bioactive Phytochemicals: Drug Discovery to Product Development Ahmad & Ahamad
R
Ratio, drug extract 134
Reaction 77, 107, 110
jumping 107
nitroso 77
postural 110
Response 191
organism’s 191
pathophysiological 191
Reversed-phase HPLC 64
Rheumatoid arthritis 192, 193
Rosmarinus officinalis 38, 41
Rotarod test 111
S
Saline 110, 113, 115
ice-cold phosphate-buffered 113
Salvia officinalis 200
Sarsaparilla officinalis 27
Saudi food and drug authority (SFDA) 160,
170
Screening 1, 2, 7, 55, 64, 73, 101, 103, 107,
109, 115, 188
bioassay 64
biological 73
high throughput 55
pre-clinical 1, 2
Selected reaction monitoring (SRM) 87, 96
Siddha system of medicine (SSM) 122, 124,
127, 159
Skeletal muscle functions 111
Skin carcinogenesis 190
Smoking 6, 195
cessation 195
deterrent 6
Soil 14, 15, 16, 17, 18, 19, 25, 45, 133, 146,
197
adhering 45
sandy 17
wet 197
Solubility 10, 34, 35, 57, 58
relative 58
Soluble 22, 146
compounds 22
extractives 146
Solvents 33, 34, 35, 36, 37, 38, 39, 40, 56, 59,
64, 65, 146, 152
eluting 64
non-polar 35
polarity of 35
Solvent systems 58
binary 58
Sophisticated isolation technique 65
Source plants solvent system 58, 59
South african health products regulatory
agency (SAHPRA) 160
Soxhlet 37, 39
extraction method 37, 39
method 37
Spectral analyses 75
Spectroscopy 73, 75, 77, 80, 85, 152
atomic absorption 152
correlation 85
Spray-drying process 26
Standard bleeding techniques 104
Staphylococcus aureus 136
Steroidal synthesis 43
Structural 74, 88
characterization of natural products 88
elucidation of phytoconstituents 74
Sugar 48, 168
digitoxose 48
Supercritical fluid extraction (SFE) 34, 37, 38,
40, 42, 49, 134
Sweetening agent 130
T
Tanzania food and drug administration
(TFDA) 160
Techniques 40, 56, 57, 58, 59, 65, 66, 73, 74,
75, 80, 84, 86, 95, 128, 153, 154
freeze-drying 128
hyphenated 73, 74, 95
of natural product isolation 56
Technology, multidimensional protein
identification 86
Tests 7, 75, 111, 113, 132, 134, 135, 136, 182
cytotoxicity 7
glucose tolerance 182
physicochemical 75
products uniformity 135
releasing 136
toxicity 7
Tetracyclic diterpene acids 21
Tetrahydrocontinentic acid 60
Therapeutic regimen 117
Subject Index Bioactive Phytochemicals: Drug Discovery to Product Development 221
Thermolabile 33, 34, 35, 36, 37, 39, 40, 65
components 40
compounds 35
drugs 36, 37
extracting 33
herbal drugs 35, 39
Thin layer chromatography 65, 133, 151, 152,
168
Thrombolysis 195
Thymus vulgaris 6
Tibetan medicine system 122
Tissues 21, 22, 40, 114, 145, 191, 192, 198
adipose 198
coleoptile 21
vascular 22, 191
Toluene distillation process 147
Topographical influence 19
Toxic residues in herbal formulation 151
Traditional 37, 124, 159
Chinese medicines (TCM) 37, 124, 159
Indian medicine (TIM) 159
Korean medicine (TKM) 159
Traditional medicine (TM) 1, 2, 4, 37, 101,
122, 137, 138, 144, 160, 161, 163, 164,
165, 177, 178
system 101, 144
Treatment of cardiovascular diseases 196
U
Ulcer 104, 112, 113
hemorrhagic 113
index (UI) 112, 113
Ultrasonic-assisted extraction (UAE) 34, 38,
39, 41, 49
United states 132, 144, 147, 160, 164, 166,
167
food and drug administration (USFDA)
132, 160, 164
pharmacopoeia (USP) 144, 147, 160, 166,
167
USP herbal medicine compendium 167
UV 42, 76
spectroscopy 76
spectrum 42
UV-visible 62, 73, 74, 75, 76, 77
absorption characteristics 77
spectroscopic detectors 62
Spectroscopy 74, 75, 76
spectroscopy and mass spectrometry 73
V
Vacuum 26, 34, 36, 37, 39, 48, 65, 87
chamber 87
dryer 26
Drying 26
Valepotriates 7
Valeriana officinalis 7
Vitali-Morin reaction 77
Volatile 25, 193
fractions 193
principles 25
W
Water 17, 18, 33, 44, 45, 58, 59, 64, 113, 129,
130, 131, 146, 147, 148
extraction 33
preparations 130
Wavelength 75, 76, 77
spectra record 75
WHO guidelines 131, 132, 133, 137, 138,
143, 145, 152, 154
World health organization (WHO) 46, 122,
124, 131, 137, 138, 144, 159, 160, 161,
164, 166, 168
X
X-ray crystallography 116
Z
Zingiber officinale 193, 198
