Experimental Pharmacognosy and Phytochemistry

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S. S Khadabadi, S. L Deore, B A Baviskar
2016, 454pp
Paperback, 9788193033364
Rs. 545

Natural drugs and derived products are gaining worldwide popularity due to numerous advantages including easy availability, possible source of lead molecules, synergism with modern drugs and their significant research opportunities. Due to the advancement in chromatographic (e.g., Flash chromatography) and spectroscopic (e.g., sensitive NMR probe analysis, RDC-Residual Dipolar Coupling) technologies, the integration of natural products in HTS (High Throughput Screening) is possible. Again the better understanding of intellectual property rights and regulatory affairs by research scientists, students and professional has increased the pace of research for technological development in herbal drugs.

Thus, the authors have designed this book to cover all important and necessary information of experimentations in pharmacognosy and phytochemistry. This book will provide readers with a concise overview of extraction, isolation, purification, qualitative and quantitative estimation methods of different natural drugs of classes like glycoside, alkaloids, volatile oils, lipids, carbohydrates, pigments and resins.

Relevant basic theory is provided to have better understanding of experiments. List of spraying and common reagents can be very helpful in day-to-day laboratory work. This is not intended to be a complete and comprehensive list, but should set the stage for future elaborations.

The book as a whole is aimed at all students, professionals to keep abreast with details of experiments in phytopharmacognosy. Authors hope that this book will be very helpful to the new as well as experienced researchers directly or indirectly concerned to this field.

Key Features

  • Unique in its coverage
  • Provides concise rewritten clear text
  • Well documented much useful laboratory techniques
  • Compressive index and innovative presentation
  • Spectroscopic details for few natural chemicals
  • Herbal cosmetic formulas
  • Plant tissue culture experiments
  • More than 50 estimation techniques
  • Isolation methods for more than 70 natural chemical compounds
  • More than 100 general chemical tests for preliminary phytochemical screening
  • More than 100 specific confirmatory tests for crude drug or pure natural compounds

Dr. S. S Khadabadi, Principal, Government College of Pharmacy, Amravati, Maharashtra.
Dr. S. L Deore, Assistant Professor, Government College of Pharmacy, Amravati, Maharashtra.
Dr. B A Baviskar, Assistant Professor, Government College of Pharmacy, Amravati, Maharashtra.

1 Extraction and Isolation Method
1.1 Extraction of Plant Drugs
1.2 Choice of Extraction Methods
1.3 Drying of Crude Drugs
1.3.1 Methods of Drying
1.4 Methods of Extraction
1.4.1 Maceration Method
1.4.2 Percolation Method
1.4.3 Hot Continuous Counter-current Extraction Method
1.4.4 Infusion Method
1.4.5 Decoction Method
1.4.6 Water Distillation Method
1.4.7 Steam Distillation Method
1.4.8 Expression Method
1.4.9 Enfleurage Method
1.4.10 Ultrasound Extraction Method
1.4.11 Microwave Assisted Extraction Method
1.4.12 Countercurrent Distribution (Craig Apparatus)
1.4.13 Droplet Countercurrent Chromatography (DCCC)
1.4.14 Accelerated Solvent Extraction (ASE) Method
1.4.15 Supercritical Fluid Extraction (SCF)
1.4.16 Solid-Phase Extraction (SPE)
1.4.17 Liquid-Liquid Extraction (LLE)
1.5 Solvent Evaporation and Drying of Extracts

2 Purification Methods
2.1 Introduction
2.2 Removal of Interfering Substances
2.3 Chromatographic Methods of Purification
2.3.1 Planar Chromatography
2.3.2 Column Chromatography
2.4 Hyphenated Techniques
2.5 Evaporation
2.6 Sublimation
2.7 Crystallization
2.8 Electrophoresis
2.8.1 Types of Electrophoresis

3 Preliminary Chemical Evaluation
3.1 Introduction
3.2 General Tests for Preliminary Phytochemical Screening
3.2.1 Preliminary Test
3.2.2 General Tests for Phytoconstituents
3.3 Confirmatory Chemical Tests for Phytoconstituents
3.4 Confirmatory Chemical Tests for Organized Crude Drugs
3.5 Confirmatory Chemical Tests for Unorganized Crude Drugs

4 Quantitative Chemical Estimations
4.1 Introduction
4.2 To Estimate Total Carbohydrate by Phenol Sulphuric Acid Method
4.3 To Estimate Total Carbohydrate by Anthrone Method
4.4 To Estimate Total Glucose by Glucose Oxidase Method
4.5 To Estimate Total Reducing Sugar by Dinitrosalicylic Acid (DNS) Method
4.6 To Estimate Total Proteins by Biuret Method
4.7 To Estimate Proteins by Bradford Method
4.8 To Estimate Proteins by Lowry Assay
4.9 To Estimate Total Phenolic Contents by Folin-Ciocalteu Method
4.10 To Estimate the Total Flavonoid by Aluminium Chloride and 2,4-Dinitrophenylhydrazine Colorimetric Method
4.11 To Estimate Crude Fibers in Given Sample
4.12 To Estimate Total Steroids in Plants
4.13 To Estimate Total Anthraquinone in Plant Material
4.14 To Estimate Total Tannins by Folin-Denis Reagent
4.15 To Estimate Total Triterpenoids in Terminalia arjuna Bark
4.16 To Separate the Plant Leaf Pigments by TLC
4.17 To Separate the Spinach Leaf Pigments by Column Chromatography
4.18 To Determine Reserpine in Rauvolfia by Photometric Method
4.19 To Estimate Quinine in Cinchona Bark
4.20 To Determine Cardio Active Principles from Thevetia Seeds/Bark Calculated in Terms of Digitoxigenin by Photometric Method
4.21 To Determine Total Carvone Content of Umbelliferous Fruits
4.22 To Determine Total Citral Content in Lemon Oil
4.23 To Estimate Total Tropane Alkaloids using UV-visible Spectroscopy
4.24 To Estimate Total Saponin in Suitable Plant Material by Gravimetric Methods
4.25 To Determine Ascorbic Acid (Vitamin C) by Colorimetric Method
4.26 To Determine Acid Value of Given Sample as per IP
4.27 To Determine Total Moisture Content of Acacia by Toluene Distillation Method as per IP
4.28 To Determine Iodine Value of Given Sample as per IP
4.29 To Determine Saponification Value as per IP
4.30 To Determine the Unsaponifiable Matter as per IP
4.31 To Determine Free Radical Scavenging Activity by In-vitro Spectrophotometric Analysis
4.32 To Determine the Presence of Rhapontic Rhubarb in Given Sample of Rhubarb by the Use of Paper Chromatography
4.33 To Estimate Total Triterpenoids by Colorimetric Method
4.34 To Estimate Solanine by UV-visible Spectroscopic Method
4.35 To Estimate Morphine by UV-visible Spectroscopic Method
4.36 To Estimate Colchicine by Ferric Chloride Method
4.37 To Estimate Total Alkaloids of Ergot by Colorimetric Method
4.38 To Estimate Brucine by Colorimetric method
4.39 To Estimate Embelin by UV-visible Spectroscopic Method
4.40 To Estimate Ephedrine by UV-visible Spectroscopic Method
4.41 To Estimate Rutin by UV-visible Spectroscopic Method
4.42 To Isolate DNA from Onion
4.43 To Determine Alcohol Content of Asava and Aristha
4.44 To Perform Microwave Assisted Extraction of Some Phytoconstituents
4.45 To Perform HPLC Analysis of Some Phytoconstituents
4.46 To Estimate Total Alkaloids in Plant Extracts
4.47 To Determine Total Bitterness Value of Given Crude Drug
4.48 To Determine Total Haemolytic Activity of Saponin Containing Crude Drugs
4.49 To Determine Total Tannins of a given Crude Drug
4.50 To Determine Foaming Index of Saponin containing Crude Drug
4.51 To Determine Swelling Index of a given Crude Drug
4.52 To Determine the Anti-Microbial Activity of Essential Oils
4.53 To Determine Microbial Load in Given Sample

5 Alkaloids
5.1 Introduction
5.2 Methods of Total Alkaloid Extraction
5.2.1 General Methods of Extraction
5.2.2 Stass-otto Process of Alkaloid Extraction
5.3 Types of Alkaloids
5.3.1 Atropine
5.3.2 Berberine
5.3.3 Caffeine
5.3.4 Colchicine
5.3.5 Coniine
5.3.6 D-tubocurarine Chloride (DTC)
5.3.7 Emetine
5.3.8 Ephedrine
5.3.9 Ergometrine (Ergonovine)
5.3.10 Ergotamine
5.3.11 Lobelia Total Alkaloids
5.3.12 Lobeline
5.3.13 Morphine and Codeine (Opium Alkaloids)
5.3.14 Nicotine
5.3.15 Physostigmine
5.3.16 Piperine
5.3.17 Quinine and Quinidine
5.3.18 Reserpine (Raudixin, Serpalan, Serpasil)
5.3.19 Solanine
5.3.20 Solasodine
5.3.21 Sparteine
5.3.22 Strychnine and Brucine
5.3.23 Taxol (Paclitaxel)
5.3.24 Vasicine (Peganine)
5.3.25 Vinblastine and vincristine (Vinca Alkaloids)

6 Glycoside Isolation and Extraction
6.1 Introduction
6.2 General Method of Total Glycoside Extraction
6.3 Pure Glycoside Extraction Method
6.3.1 Aloe Gel
6.3.2 Aloin (Barbaloin)
6.3.3 Andrographolide
6.3.4 Bacosides
6.3.5 Boswellic Acids
6.3.6 Citric Acid
6.3.7 Digoxin (Lanoxin)
6.3.8 Diosgenin
6.3.9 Ginsenosides
6.3.10 Guggulsterone
6.3.11 Gymnemic Acids
6.3.12 Glycyrrhetinic Acid (Enoxolone)
6.3.13 Hesperidin
6.3.14 Hecogenin
6.3.15 Hypericin
6.3.16 Naringin
6.3.17 Papain
6.3.18 Psoralen
6.3.19 Rhein
6.3.20 Rutin (Rutoside, Quercetin-3-O-Rutinoside and Sophorin)
6.3.21 Sennosides
6.3.22 Stigmasterol
6.3.23 Tannic Acid
6.3.24 Withanolides

7 Volatile Oils
7.1 Introduction
7.2 Extraction, Isolation and Purification Methods
7.2.1 Dry Distillation Method
7.2.2 Solvent Extraction Method
7.2.3 Rectification
7.2.4 General Method of Extraction of Volatile Oils
7.3 Analysis of Volatile Oils
7.4 Specific Volatile Oils
7.4.1 Artemisinin
7.4.2 Asarone
7.4.3 Azadhirictin
7.4.4 Camphor
7.4.5 Eugenol
7.4.6 Forskolin
7.4.7 Menthol

8 Plant Pigments
8.1 Introduction
8.2 Different Classes of Pigments
8.3 Extraction of Plant Pigments
8.3.1 Bixin
8.3.2 Capsanthin
8.3.3 Curcumin
8.3.4 Lawsone
8.3.5 Lycopenes

9 Lipids
9.1 Introduction
9.2 Extraction Method of Lipids
9.2.1 Two-Step Bligh and Dyer Method
9.3 Analysis of Lipids
9.3.1 Methylation of Fatty Acids
9.3.2 Simultaneous TLC and Esterification
9.3.3 Direct Transesterification Method
9.4 Extraction of Selected Lipids
9.4.1 Cetyl Alcohol (1-hexadecanol and Palmityl Alcohol)
9.4.2 Myristicin

10 Resins
10.1 Introduction
10.2 Isolation and Purification of Different Resin Chemical Moieties
10.2.1 General Extraction Method
10.2.2 Abietic Acid
10.2.3 Cannabinoids
10.2.4 Capsaicin
10.2.5 Gingerols and Shagaols
10.2.6 Jalap Resin
10.2.7 Laccaic Acids (Hydroxyl Anthraquinone Carboxylic Acid)
10.2.8 Podophyllotoxin

11 Carbohydrates
11.1 Introduction
11.2 Extraction of Carbohydrates
11.2.1 General Method of Extraction
11.2.2 Agar
11.2.3 Cellulose
11.2.4 Mucilage
11.2.5 Pectin
11.2.6 Starch (Amylopectin and Amylase)

12 Plant Tissue Culture
12.1 Introduction
12.2 Basics of Plant Tissue Culture (PTC)
12.2.1 Totipotency
12.2.2 Somatic Embryogenesis
12.2.3 Organogenesis
12.3 Requirements for Establishment of PTC
12.3.1 Plant Tissue Culture Lab
12.3.2 Explant Selection and Surface Sterilization
12.3.3 Plant Tissue Culture Media
12.3.4 Callus Culture
12.3.5 Suspension Culture
12.4 Growth Profile for Plant Tissue Culture
12.5 Plant Culture Growth Measurement
12.6 Types of Culture
12.7 Plant Tissue Culture Applications
12.8 Examples of PTC
12.9 Laboratory Exercise
12.9.1 To Prepare Murashige and Skoog (MS) Medium
12.9.2 To Isolate and Sterilize Explant for Callus Culture
12.9.3 To Immobilize Plant Cells by Gel Entrapment Method
12.9.4 To Prepare Protoplasts from Young Leaves
12.9.5 To Develop Meristem Culture of Potato
12.9.6 To Develop Haploid Plants from Anther Culture
12.9.7 To prepare Synthetic Seeds
12.9.8 To Develop A. tumefaciens Mediated Transformation in Plants
12.9.9 To Initiate Organogenesis in Tomato Nodal Explants
12.9.10 To Initiate Somatic Embryogenesis in Aloe
12.9.11 To Initiate Micropropogation of Citrus Through Seeds

13 Cosmetics
13.1 Introduction
13.2 Need of Cosmetics
13.3 Classification
13.4 Evaluation of Cosmetics
13.5 Herbal Cosmetics
13.6 Laboratory Exercise
13.6.1 To Prepare Herbal Cold Cream
13.6.2 To Prepare Herbal Cleansing Cream
13.6.3 To Prepare Herbal Face Packs
13.6.4 To Prepare Herbal After Shave Lotions
13.6.5 To Prepare Herbal Sunscreens Cream
13.6.6 To Prepare Herbal Talcum Powder
13.6.7 To Prepare Herbal Lipsticks
13.6.8 To Prepare Herbal Shampoo
13.6.9 To Prepare Herbal Hair Conditioner
13.6.10 To Prepare Herbal Tooth Powder
13.6.11 To Prepare Herbal Mouth Wash

14 Spectroscopic Elucidation of Natural Compounds
14.1 Introduction
14.2 Fundamental Laws of Spectrophotometry
14.3 General Principle
14.4 Types of Spectroscopy
14.4.1 Ultraviolet (UV) and Visible Spectroscopy
14.4.2 Infrared (IR) Spectroscopy
14.4.3 NMR Spectroscopy
14.4.4 Mass Spectroscopy
14.5 Spectroscopic Elucidation of Phytoconstituents
14.6 Index of Hydrogen Deficiency (IHD)
14.7 Chemical and Spectroscopic Details of Some Natural Compounds
14.7.1 Caffiene
14.7.2 Morphine
14.7.3 Atropine
14.7.4 Reserpine
14.7.5 Cholesterol
14.7.6 Diosgenin
14.7.7 Stigmasterol
14.7.8 Sitosterol
14.7.9 Lupeol
14.7.10 Gallic Acid
14.7.11 Quercetin
14.7.12 Rutin
14.7.13 Myristic Acid
14.7.14 Linoleic Acid
14.7.15 Glucose
14.7.16 Glactose

15 Morphology and Microscopy of Crude Drugs

15.1 Morphological Evaluation
15.2 Microscopical Evaluation
15.3 Laboratory Exercise
15.3.1 To Study Microscope
15.3.2 To Study General Techniques Involved in Section Cutting, Staining, Mounting and Micro Chemical Testing
15.3.3 To Prepare Herbarium of Plants
15.3.4 To Perform Microtome Section Cutting of Crude Drugs by Rotary Microtome
15.3.5 To Determine Palisade Ratio of Given Leaf Crude Drug
15.3.6 To Determine Vein-Islet Number of Given Leaf Crude Drug
15.3.7 To Determine Vein-Termination Number of Given Leaf Crude Drug
15.3.8 To Determine Stomatal Number and Stomatal Index of Given Leaf Crude Drug
15.3.9 To Determine Percentage Purity by Lycopodium Spore Method
15.3.10 To Perform Powder Microscopic Evaluation Of Given Crude Drug
15.3.11 To Calibrate Eyepiece Micrometer by Stage Micrometer
15.3.12 To Measure Dimensions Of Starch Grain in a Given Crude Drug Powder
15.3.13 To Measure Length of Phloem Fibers in Given Crude Drug Powder
15.4 Morphology and Microscopy of Crude Drugs
15.4.1 Caraway
15.4.2 Cascara
15.4.3 Cassia
15.4.4 Cinchona
15.4.5 Cinnamon
15.4.6 Clove
15.4.7 Coriander
15.4.8 Digitalis
15.4.9 Ephedra
15.4.10 Eucalyptus
15.4.11 Fennel
15.4.12 Gentian
15.4.13 Ginger
15.4.14 Ipecac
15.4.15 Ispaghula
15.4.16 Kalmegh
15.4.17 Kurchi
15.4.18 Liquorice
15.4.19 Nux Vomica
15.4.20 Quassia
15.4.21 Rauwolfia
15.4.22 Rhubarb
15.4.23 Senna
15.4.24 Squill
15.4.25 Stramonium
15.4.26 Valerian
15.4.27 Vasaka
15.4.28 Vinca

16 Alternative Systems of Medicine

16.1 Ayurvedic System
16.1.1 Fundamental Principles
16.1.2 Diagnosis and Treatment
16.1.3 Preparation and Evaluation of Ayurvedic Formulations
16.2 Homeopathy System
16.2.1 Fundamental Principles
16.2.2 Diagnosis and Treatment
16.3 Chinese System
16.3.1 Fundamental Principles
16.3.2 Diagnosis and Treatment
16.4 Siddha System
16.4.1 Fundamental Principles
16.4.2 Diagnosis and Treatment
16.5 Unani System
16.5.1 Theory and Principles
16.5.2 Unani System Dosage Forms
16.5.3 Diagnosis and Treatment
16.6 Quality Control Parameters for Traditional Medicines

Annexure 1 Reagents
Annexure 2 Thin Layer Chromatography Details of Phytoconstituents
Annexure 3 Solvents and their Properties

Further Reading