Quality Control and Evaluation of Herbal Drugs: Evaluating Natural Products and Traditional Medicine

Cover
Quality Control and
Evaluation of Herbal Drugs:

Evaluating Natural Products and
Traditional Medicine
Copyright
Preface
Foreword
	Good Affordable Evidence-Based (Herbal) Medicines for Everyone
Acknowledgments
1
Traditional Systems of Medicine and Harmonization
	Traditional Medicine in Healthcare
	AYUSH Systems of Medicine
		Traditional Knowledge Digital Library
	Ayurveda
		Origin of Ayurveda
		Ayurvedic Drugs and Pharmaceuticals
		Medicinal Plants in Ayurveda
		Globalization of Ayurvedic Medication
		Ayurvedic Treatment and Practice
	Unani
		Therapy and Formulation of Unani Medicine
	Siddha
	Homeopathy
	Sowa-Rigpa
	Traditional Chinese Medicine
		Concepts of Traditional Chinese Medicine
		Quality Evaluation of Chinese Herbal Medicine
	Japanese Traditional Medicine
	Thai Herbal Medicine
	Traditional Medicine in Bangladesh
	Iranian Traditional Medicine
	Malaysian Traditional Medicine
	Traditional Medicine in Australia
	Nature’s Natural Product Library for New Drug Discovery
	Drugs Derived From TM
	Harmonization of Traditional Systems and Health Practices
	References
	Further Reading
2
Ethnopharmacology and Ethnomedicine-Inspired Drug Development
	Ethnopharmacology and Integrative Medicine
	Ethnobotany in Drug Development
		Ethnobotany in Traditional Medicine and Modern Therapy
		Drug Discovery From Ethnobotany
		Ethnobotany and Drug Development
	Anthropological Aspects of Ethnopharmacology
	Ethnomedicine in Healthcare
	Ethnomedicine in Complementary Therapeutics
	Validation of Ethnomedicines
	Ethnopharmacology and Translational Research
	Biological Wealth of Ancient Wisdom
	Approaches to Drug Development Inspired by Traditional Medicine
	Intellectual Property (IP) Rights in Ethnopharmacology
	Conservation of Biodiversity in Ethnopharmacology
		Ethnobioresources and Conservation Approaches
	Network Pharmacology in Ethnopharmacological Research
	Reverse Pharmacology
	Phytoequivalence
	Phytopharmaceuticals
	References
	Further Reading
3
Quality Evaluation of Herbal Medicines: Challenges and Opportunities
	Factors Affecting the Quality of Herbs
		Authentication and Reproducibility of Herbal Ingredients
		Inter/Intra Species Variation in Plants
		Environmental Factors
			Solar Radiation
			Soil Nutrients
			Stress
				Water Stress
				Drought Stress
				Salinity Stress
				Chemical Stress
				Temperature
				Ecological Factors
			Seasonal Variation
		Geographical Location
		Plant Parts Used
		Time of Harvesting
		Post harvesting Factors
		Contaminants in Herbal Ingredients
			Environmental Contaminants
			Pesticides, Fumigants, and Other Toxic Metals
		Chemical Variation in Medicinal Plants
	Adulterations of Herbal Drugs
		Various Means of Adulterations in Herbal Medicine
			Substitution With Inferior Commercial Varieties
			Adulteration by Artificially Manufactured Substitutes
			Substitution by Exhausted Drugs
			Substitution by Superficially Similar but Cheaper Natural Substances
			Adulteration by Addition of Worthless Heavy Materials
			Addition of Synthetic Principles
			Adulteration With Inferior Drugs
			Harmful Adulterants
		Reasons of Adulteration
			Confusion in Vernacular Names
			Lack of Knowledge About Authentic Sources
			Similarity in Morphology
			Similarity in Color
			Careless Collections
			Other Unknown Reasons
	Deterioration of Herbal Drugs
		Light
		Moisture
		Temperature
		Airic Oxidation
		Bacteria and Molds
		Mites and Nematode Worms
		Insects and Moths
		Coleoptera or Beetles
		Control Measures for Deterioration
	Substitution of Herbs
		Substitution of the Species Belonging to the Same Family
		Substitution of Different Species
		Substitution of Different Parts of the Same Plant
		Substitution Due to Same Action
	Counterfeiting of Herbal Medicine
		Types of Counterfeit Drugs and Their Consequences
		Preventive Measures
		Methods of Detection
	Collection of Herbs
		Methods of Bark Collection
			Felling Method
			Uprooting Method
			Coppicing Method
		Illustrative Examples With Discussion
		Good Agricultural Practices
	Identification and Authentication of Herbs
	Taxonomy of Herbs
		Nomenclature of Medicinal Plants
		Author Names and Synonyms
		Classification Systems
	Herbarium Documentation for Medicinal Herbs
		Voucher Specimen
		Preparation of Herbarium
			Collection of Plant Material
			Pressing and Drying
			Labeling of Specimen
			Storage of Specimens
	References
	Further Reading
4
Qualitative Analysis for Evaluation of Herbal Drugs
	Importance of Qualitative Analysis
	Moisture Content
		Importance and Significance of Moisture Content
		Determination of the Moisture Content of a Crude Drug
			The Azeotropic Or Toluene Distillation Method
			Determination of the Loss on Drying
			Titrimetric Determination of Moisture Content
	Ash Values
		Principles of Determination of Ash Value
		Methodology of Determination of Ash Value
			Dry Ashing Technique for Total Ash Determination
			Wet Ashing Technique for Total Ash Determination
			Acid-Insoluble Ash
			Water-Soluble Ash
			Determination of Sulfated Ash
			The Microwave Ashing Technique
			Dry Ashing Using a Microwave System
		Comparative Evaluation of Different Ashing Techniques
		Near-Infrared Spectroscopy for the Determination of the Ash Value
	Determination of Extractive Value
		Principles and Methodology of the Determination of Extractive Values for Medicinal Plants
		Procedures for Determination of Extractive Values
			Determination of Water-Soluble Extractive
			Determination of the Ethanol-Soluble Extractive
			Determination of Hexane-Soluble Extractive
			Volatile Ether-Soluble Extractive
			Nonvolatile Ether-Soluble Extractive
	Residual Solvents
		Classification of Residual Solvents
			Class 1 Solvents: Solvents to Be Avoided
			Class 2 Solvents: Solvents to Be Limited
			Class 3 Solvents: Solvents With Low Toxic Potential
	Total Solids
		Significance and Use
	Determination of Crude Fiber
	Bitterness Determination
		Procedure for Determination of Bitterness
	Swelling Index
		Determination of Swelling Index
	Foaming Index
		Determination of the Foaming Index
			Method 1
			Method 2
			Foam Test
	Hemolytic Activity
		Determination of Hemolytic Activity
			Method 1
			Method 2
	Total Tannin Content
		Hydrolysable Tannins
			Gallotannins
			Ellagiotannins
		Complex Tannins
			Condensed Tannins
		Pseudotannins
		Determination of the Tannin Content (WHO, 1998)
			Titrimetric Method
			Spectrophotometric Method
			Alkaline Ethanol Precipitation Method
		The Tannin Content of Several Medicinal Plants
	Total Phenolic Contents
	Total Flavonoid Contents
		Method 1
		Method 2
		Method 3
	Heavy Metals
		Toxic Elements
		Effects of Heavy Metals on Secondary Metabolites
		Atomic Absorption Spectrometry (AAS)
			Sample Preparation
		Instrumentation
		Techniques Based on Atomic Absorption Spectrometry
			Calibration Curve Method
			Standard Addition Method
			Internal Standard Method
		Limit Test for Arsenic
			Preparation of the Sample by Acid Digestion
			Apparatus and Methodology
			Preparation of Standard Stain
			Limit Test for Cadmium and Lead
				Apparatus
			Preparation of the Sample
				Method
		Limit Test for Mercury
			Materials and Methods
			Apparatus
			Standard Preparation
			Analysis Method
	Pesticides
		Pest Control and Pesticide
		Chemical Pest Control
		Different Pesticides and Chemical Composition
		Various Forms of Preparations of Pesticides Used
			Classification of Pesticides
				Fungicides
				Herbicides
				Insecticides
				Acaricides
				Nematocides
				Rodenticides
				Bactericides
		Adverse Effects of Different Pesticides
		Maximum Limit of Pesticide Residues for Herbal Drugs
		Limits for Pesticides
		Pests and Pest Control
			Microbes
			Insects
			Noninsect Pests
			Weeds
		Different Classes of Pesticide Based on Chemical Composition
		Method for Determination of Pesticides
			Method 1
			Method 2
			Method 3
			Method 4
			Method 5
	Aflatoxins and Other Microbial Contents
		Factors Affecting Microbial Content
		Microbial Quality Standards of WHO
		Principles for the Determination of Microbial Load
		Procedures for the Determination of Microbial Load
			Total Aerobic Viable Count
			Growth Promotion by Media
			Appropriateness of the Enumeration Method in the Presence of Product
			Membrane Filtration Method
			Pour Plate Method
			Most Probable Number (MPN) Method
		Examples of the Importance of Microbial Load According to the Indian Pharmacopeia
		Detection of the Presence of Specific Microorganisms
			Escherichia coli
			Salmonella Species
			Pseudomonas Species
			Candida albicans
		Determination of Aflatoxins
	Essential Oil in Crude Drugs
		Principles and Methods for Determination of Essential Oil Content
			Distillation
			Hydrodiffusion
			Water Distillation
			Water and Steam Distillation
			Direct Steam Distillation
			Cohobation
			Hydrodistillation Through Clevenger Apparatus
			Microdistillation Method
		Simultaneous Extraction Coupled to TLC
		Direct Sampling From the Secretory Structures
		Supercritical Fluid Extraction
		Microwave-Assisted Extraction
		Illustrative Example of the Determination of the Essential Oils in Crude Drugs
		Preliminary Quality Control Parameters for Essential Oils
			Total Eugenol Content
			Determination of Total Cineole Content
			Determination of Total Aldehyde Content in Volatile Oil
			Determination of the Carvone Content of Volatile Oil
	References
	Further Reading
5
Morphological and Microscopical Evaluations
	Organoleptic Evaluation
		Attributes of Organoleptic Evaluation
			Color
			Size
			Odor and Taste
			Surface Characteristic: Texture and Fracture
	Macromorphological Evaluation
	Macromorphology of Crude Drugs From Barks
		Nature of Curvature
		Surface Characteristics
		Characteristics of Fracture
		Characteristics of Transverse Surface
		Histology of Barks
	Macromorphology of Crude Drugs From Roots
		Modification for Storage of Food
		Modifications for Support
		Modifications for Special Functions
	Macromorphology of Crude Drugs From Stems
		Underground Stem
		Stem Tendrils
		Thorn
		Aerial Stem
		Weak Stems
		Subaerial Weak Stem
		Herbaceous and Woody Stems
	Macromorphology of Crude Drugs From Rhizomes
		Occurrence or Form of the Rhizome
		Shape of the Drug
		Surface Characters
		Transverse Section
	Macromorphology of Crude Drugs From Woods
		Size and Color
		Fracture and Hardness When Split
		Transverse Surface
	Macromorphological Description of the Entire Aerial Parts
	Macromorphology of Crude Drugs From Leaves
		Types of Leaves
		Structural Distribution of Leaves
		Arrangement and Position of Leaves
		Surface Appearance and Texture
	Macromorphological Description of Herbal Drugs Containing Flowers
		Receptacle of Floral Drugs
		Characteristics of Inflorescence
		Arrangement of Floral Parts on Thalamus
	Macromorphological Description of Fruit Drugs
		Shape and Size
		Types of Fruits
	Macromorphological Evaluation of Crude Drugs Obtained From Seeds
		Shape of the Seeds
		Seed Coat
		Special Features of Seeds
	Macromorphological Evaluation of Cellular Materials or Unorganized Drugs
		Evaluation of Physical State
		Vegetable Debris
		Effect on Heating
		Morphological Characteristics of a Few Unorganized Herbal Drugs
	Cytomorphological Evaluation of Herbal Drugs
		Analytical Application of Cytomorphology
			Epidermis
			Stomata
			Trichomes
		Basic Cell Types and Cell Inclusions Present in Herbal Drugs
			Parenchyma
			Sclerenchyma
			Collenchyma
			Xylem
			Phloem
			Epidermis
			Periderm
			Secondary Tissues
			Calcium Oxalate
			Calcium Carbonate
			Starch
	Microscopical Techniques
		Instruments for Microscopical Study
			Camera Lucida
			Photomicrography
			Modified Light Microscopy
			Polarizing Microscopy
			Phase Contrast Microscopy
			Electron Microscopy
			Ultraviolet Microscopy
		Microchemical Testing of Herbal Drugs
		Microchemical Precipitation
		Micro-sublimation
	Evaluation of Crude Drugs by Microscopy
		Preparation of Crude Drug Sample for Microscopical Examination
		Clearing Reagents and Preliminary Treatment
			Chloral Hydrate Solution
			Phenols
			Nitric Acid
			Hydrochloric Acid
			Hydrofluoric Acid
			Ether–Ethanol Reagent
			Sodium Hypochlorite Solution
			Caustic Alkali
			Clove Oil
	Quantitative Analytical Microscopy
		Optical Micromeritics
		Micrometers
		Micrometry
		Leaf Characters for the Evaluation of Impurity
			Vein Islet and Veinlet Termination
			Stomatal Index
			Palisade Ratio
			Lycopodium Spore Method
	References
6
Extraction and Other Downstream Procedures for Evaluation of Herbal Drugs
	Basic Principles and Rationale
	Factors Affecting Extraction of Herbal Drugs
		Choice of Solvents
		Types of Extracts
		Raw Materials
		Choice of Solvent
		Conditions for Extraction
		Selection, Collection, and Identification of Plant Material
		Drying of the Crude Drugs
		Comminution and Classification
			Medicinal Leaves and Herbs
			Roots and Barks
			Seeds and Fruits
	Methods of Extraction
		Factors Affecting the Choice of Extraction Process
		Maceration, Digestion, and Remaceration
			Advantages and Disadvantages
			Percolation and Repercolation
			Examples of Experiments on Percolation and Observations
			The Factors Affecting the Percolation Process
		Infusion and Decoction
		Vortical or Turbo Extraction
		Ultrasound Extraction
		Extraction by Electrical Energy
		Countercurrent Extraction
			Continuous Countercurrent Extraction
			Relative Countercurrent Extraction
		Hot Continuous Extraction—Soxhlation
			Advantages of a Conventional Soxhlet Apparatus
			Disadvantages of a Conventional Soxhlet Apparatus
			High-Pressure Soxhlet Extraction
			Automated Soxhlet Extraction
			Ultrasound-Assisted Soxhlet Extraction
			Microwave-Assisted Soxhlet Extraction
		Extraction of Essential Oil
			Distillation
			Hydro Diffusion
			Water Distillation
			Water and Steam Distillation
			Direct Steam Distillation
			Cohobation
		Microwave Extraction
			Principles
			Microwave-Assisted Subcritical and SFE
			Microwave-Assisted Enzymatic Extraction
		Supercritical Fluid Extraction
		Pressurized Liquid Extraction
			Illustrative Example
			Pressurized Hot Water Extraction (PHWE)
		General Protocols for Extraction of Phytoconstituents
	Effect of Solvent, Solvent Mixtures, and Solution on Extraction
		Solvents Used for Extractions
		Extraction Enhancement by Emerging Extraction Solvents
			Ionic Liquids
			Other Enhancements
		Properties of Solutions
		Types of Solutions
			Azeotropic Mixtures
			Solubility of Active Constituents
			The Phase Rule
			Solubility Expressions
			Solvent–Solute Interactions
		Classification of the Solvents
			Polar Solvents
			Semipolar Solvents
			Nonpolar Solvents
		Influence of Solvents
		Combined Effect of pH and Solvents
		Examples of Effects of Solvents on the Extraction
	Characteristics of Phytoconstituents
		Polarity
		pH
		Thermostability
	Interfering Compounds in Extraction of Desired Phytoconstituents
		Lipids
		Colors and Pigments
		Plasticizers
		Water Soluble Polysaccharides and Other Carbohydrate Polymers
	Extraction Methods for Specific Phytochemical Groups
		Alkaloids
		Carotenoids
		Fixed Oils, Fats, and Waxes
		Glycosides
		Phenolic Compounds
		Proteins
		Polysaccharides
	Treatment of Drug Residue After Extraction–Downstream Processing
		Sanitization of Extracts and Elimination of Solvents
		Removal of Solvents
			Rotary Vacuum Evaporator
			Tube Vaporizers
			Falling Film Vaporizer
		Freeze Drying
		Reverse Osmosis
		Particle Size Reduction
	References
	Further Reading
7
Bioactive Phytocomponents and Their Analysis
	Bioactive Phytocomponents and Their Classification
		Classification of Bioactive Phytocomponents
	Importance of Phytoconstituents in Therapy
	Qualitative Analysis of Crude Drug Extracts and Isolates
		Tests for Alkaloids
		Tests for Tannins
		Test for Glycosides
			Tests for Anthraquinone Glycosides
			Tests for Steroid Glycosides
			Tests for Cardiac Glycosides
			Tests for Coumarin Glycosides
			Tests for Cynophoric Glycoside
			Tests for Flavonoid Glycoside
		Test for Flavonoids
		Tests for Phytosterols
		Tests for Triterpenes
		Test for Saponins
		Other Polyphenols
		Tests for Resins
		Tests for Fixed Oils and Fats
		Tests for Carbohydrates
			Reagents
			Identification of Carbohydrates
			Some Distinctive Qualitative Chemical Analyses of Monosaccharides
			Some Distinctive Qualitative Chemical Analyses of Disaccharides
			Some Distinctive Qualitative Chemical Analyses of Polysaccharides
		Test for the Presence of Carotenoids
	Alkaloids and Their Analysis
		Importance of Alkaloids in Phytotherapy
		Classification of Alkaloids
			Chemical Classification
			Distribution of Alkaloids in Plant Kingdom
			Biosynthetic Classification of Alkaloids
		Pyridine–Piperidine Alkaloids
			Areca
			Lobelia
			Nicotine
		Tropane Alkaloids
			Belladonna
			Solanaceous Alkaloids
			Coca Alkaloids
		Quinoline Alkaloids
			Cinchona Alkaloids
		Isoquinoline Alkaloids
			Ipecac
			Sanguinaria
			Hydrastis
			Tubocurarine Chloride
			Opium
		Indole Alkaloids
			Rauwolfia serpentina
			Yohimbine
			Catharanthus Alkaloids
			Nux vomica
			Physostigmine
			Ergot
		Imidazole Alkaloids
			Pilocarpine
		Steroidal Alkaloids
			Veratrum viride
		Alkaloidal Amines
			Ephedrine
			Colchicine
		Purine Bases
			Caffeine-Containing Drugs
			Theophylline
			Theobromine
	Volatile Oils, Spices, and Their Analysis
		Chemical Nature of Volatile Oils and Their Analysis
		Hydrocarbons Containing Oils
			Unsaturated Acyclic Hydrocarbons
			Aromatic Hydrocarbons
			Alicyclic Hydrocarbon
		Oils Containing Alcohol
			Saturated Aliphatic Alcohols
			Unsaturated Aliphatic Alcohols
		Oils Containing Cineole
			Estimation of Cineole
			Oils of Therapeutic Importance
		Oils Containing Aldehydes
		Volatile Oils Containing Ketone
		Oils Containing Phenol
			Oils of Therapeutic Importance
		Oils Containing Esters
		Estimation of Oil of Mustard
		Physical Characteristics of Volatile Oils
		Qualitative and Quantitative Analysis of Volatile Oils
	Fixed Oils, Fats, and Waxes
		Fixed and Volatile Oils
		Chemical Nature of Fixed Oils, Fats, and Their Analysis
			Fixed Oils of Therapeutic Importance
			Fats of Therapeutic Importance
		Main Characteristic Features of Fixed Oils
	Phenylpropanoids and Their Analysis
		Classification of Phenylpropanoids
		Individual Classes of Phenylpropanoids With Examples
	Flavonoids
		Classification
			Flavanols
			Flavanones
			Isoflavones
			Flavones
		Examples of Classes of Flavonoids
			Rutin
			Apigenin
			Milk Thistle
			Quercetin
			Hesperetin
	Resins and Resinous Plant Drugs and Their Analysis
		Resins and Their Importance in Phytotherapy
			Podophyllum
			Etoposide
			Teniposide
		Chemical Classes of Resins and Their Analysis
		Quantitative Chemical Analysis of Some Resinous Compounds
	Tannins of Therapeutic Importance and Their Analysis
		Chemical Classification of Tannins and Their Quantitative Analysis
	Terpenoids and Their Analysis
		Monoterpenoids
		Sesquiterpenoids
		Diterpenoids
		Triterpenoids
		Tetraterpenoids and Carotenoids
	Glycosidal Components and Their Analysis
		Chemical Nature of Glycosides and Their Analysis
		Anthraquinone Glycosides
			C. sagrada or Rhamnus purshiana
			Frangula or Buckthorn Bark
			Aloe
			Rhubarb, Rheum, or Chinese Rhubarb
			Senna Leaf
		Saponin Glycosides
			Dioscorea
			Licorice
			Ginseng
		Cyanogenic Glycosides
		Isothiocyanate Glycosides
			Other Organosulfur Drugs
				Garlic
		Alcohol Glycosides
		Aldehyde Glycosides
		Phenol Glycosides
	Fluorescent Substances and Their Analytical Parameters
	Coloring Matters and Their Analysis
		Plastid Pigments in Plants and Their Analysis
		Pigments of the Cell Saps in Plants
			Anthoxanthins
			Anthocyanins
	Proteinous Components and Their Analysis
		Therapeutically Important Plant Proteins and Their Analysis
	Steroids
		Sterols
		Steroidal Compounds of Therapeutic Importance and Their Analysis
			Steroidal Saponins and Cardiac Glycosides
			Other Cardioactive Drugs
		Quantitative Test Procedure and Assay Methods for Cardioactive Drugs
	Carbohydrates and Their Analysis
		Classification
		Inulin
		Cellulose
		Agar
		Irish Moss and Iceland Moss
		Pentosans
		Polyuronides
		Pectic Substances
		Gums
		Acacia
		Tragacanth
		Mucilages
	References
	Further Reading
8
Thin-Layer Chromatography for Evaluation of Herbal Drugs
	Principles of TLC
	Separation of Phytoconstituents in TLC
		Efficiency of TLC System
		Resolution of TLC System
		Separation Number of TLC System
		Adsorption Chromatography
		Partition Chromatography
		Size-Inclusion/Exclusion Chromatography
		Ion Exchange Chromatography
	Adsorbents Used for TLC
		Silica Gels
		Aluminum Oxides
		Kieselguhr and Other Similar Compounds
		Cellulose
		Polyamides
		Surface-Modified Sorbents
		Hydrophobically Modified Silica Gels
		Hydrophilically Modified Silica Gels
		Surface-Modified Cellulose
		Chemical Characters
			Grain and Pore Characteristics
			Self-Made Layers
			Precoated TLC Layers
			Silica Gel Bonded Phases
	Preparation of the Layers
		Activation
		Storage, Prior to Cleaning
	Application of Samples
		Sample Application
		Preparation of Plates
		Application Methods and Equipment
	Determination of Suitable Separation Conditions
		Parameters Influencing Separation in TLC
		Selection of Developing Systems
	Development of Chromatogram
		The Solvent Characteristics
		Chamber Saturation
		Types of Development
			Ascending (Linear)
			Horizontal (Linear)
			Two Dimensional (Linear)
			Radial (Circular From the Inside Outward)
			Radial (Circular From the Outside Inward)
			Continuous Development (Overrun, Linear, and Circular)
			Multiple Development (Linear, Circular)
			AMD (Automatic Multiple Development)
		Types of Chambers and Development Techniques
			Twin Trough Chamber
			Sandwich Chamber (S-Chamber)
			Horizontal Chamber (Linear Chamber)
			Vario Chambers
			SB/CD Chamber (Regis)
			AMD Chamber (CAMAG)
		Development of Solvent System
	Detection of Herbal Drugs by TLC
		Detection by UV light
		Detection With Spray Reagents/Derivatization
	Evaluation of the TLC Plates
		Qualitative Evaluation
		Semiquantitative Evaluation
		Quantitative Evaluation
			Indirect Quantitative Evaluation
			Direct Quantitative Evaluation
	Bioautography
		Overlay Assay by TLC Bioautography
	Quantitative Thin-Layer Chromatography (QTLC)
		Quality of Sorbents for QTLC
		Demands on the Quality of Precoated Layers for QTLC
	Application of TLC for Assay of Steroids, Alkaloids, etc.
		Determination of Active Ingredient Content
		Stability Assay of Steroid Preparations
		Purity Tests of Steroid Formulated Products
		Content Uniformity Tests of Steroid-Containing Tablets and Capsules
		Problems in QTLC Determinations of Steroids
			Stability of Steroids During Analytical Procedures
			Separation Problems
			Detection Problems
		Assay of Alkaloids
			Introduction
			General QTLC Methods for Alkaloids
			QTLC Methods of the Alkaloid Groups
				Alkaloids of Ornithine and Lysine Origin
				Alkaloids of Tyrosine and Phenylalanine Origin
				Alkaloids of Tryptophan Origin
				Alkaloids of Anthranilic Acid Origin
				Alkaloids of Histidine Origin
				Steroidal Alkaloids
				Xanthine Alkaloids
				Miscellaneous Groups
	TLC Used for Some Bioactive Materials
		Case Study 1
		Case Study 2
		Case Study 3
		Case Study 4
		Case Study 5
		Case Study 6
		Case Study 7
		Case Study 8
		Case Study 9
		Case Study 10
	References
	Further Reading
9
High-Performance Thin-Layer Chromatography (HPTLC) for Analysis of Herbal Drugs
	High-Performance Thin-Layer Chromatography (HPTLC)
		Advantages of HPTLC
		Main Features of HPTLC
	Methodology in HPTLC
		Instrumentation
			LINOMAT
			Automatic TLC Sampler (ATS)
		Selection of Mobile Phase
		Preparation of HPTLC Plate
		Sample Preparation and Application
			Sample Application in HPTLC
			Sample Application in the Form of Bands or Rectangles
			Development of Plate
			Processes in the Developing Chamber
			Important Aspects to be Considered in the Development Process
				Influence of Temperature on TLC Analysis
				Influence of Coating Materials and Plates on the Performance of TLC
					Single-Beam Mode
					Double-Beam Mode
					Single-Beam Dual-Wavelength Mode
					Slit Scanning/Linear Scan
					Meander/Flying-Spot Scanning
		Retardation Factor
		Detection and Densitometric Scanning
			Derivatization and Detection
			Opto-Mechanical Scanning
			Electronic Scanning
			Laser Scanning
			Detection Methods Used in Scanning Densitometry
			Type of Scanning Mechanisms
			In Situ Quantitative Evaluation
				Baseline Compensation
				Computer-Controlled Multiwavelength Evaluation
				Photodiode Array Scanning
					Slit Width of the Monochromator
					Macro and Microparameters for the Slit Dimensions
			Parameters of the Scanner Affecting the Performance of Detection
	HPTLC Validation Procedure
		Selectivity
		Sensitivity
		Precision
		Accuracy
		Ruggedness
		Limit of Detection
		Stability
	Use of HPTLC for Standardization of Herbs and Extracts
		Simultaneous Measurement of Andrographolide and Wedelolactone
		Quantification of Eugenol, Luteoline, Ursolic Acid, and Oleanolic Acid in Black and Green Varieties of Ocimum s ...
		Estimation of Phyllanthin and Hypophyllanthin
		Determination of Curcumin
		Determination of Podophyllotoxin
		Determination of Hyperforin in Hypericum perforatum
		Determination of Corosolic Acid
		Determination of Hecogenin from Agave americana
		Quantification of Valerenic Acid in Valeriana jatamansi and Valeriana officinalis
		Analysis of Quercetin, Rutin, and Coumaric Acid in Flowers of Rhododendron
		Determination of Lycorine in Amaryllidaceae Plant Extracts
		Quantification of Fraxin in Fraxinus excelsior
		Quantification of Hypericin in Valena N Extract with St. John’s Wort
		Quantification of Commercial Ginseng Medicine
		Determination of 6-Gingerol in Zingiber officinale
		Determination of Gallic Acid in “Triphala”
		Determination Glycyrrhizin in Glycyrrhiza glabra
		Determination of Taraxerol From Coccinia grandis
		Determination of Betulinic Acid in Nelumbo nucifera
		Determination of Taraxerol in Clitoria ternatea
	Case Studies of HPTLC With Some Specific Medicinal Plants
		Withania somnifera
		Eugenia jambolana
		Ocimum sanctum
		Piper longum
		Nelumbo nucifera
		Andrographis paniculata
		Cucurbita pepo
		Glycyrrhiza glabra
		Messua ferra
		Swertia chirata
	References
	Further Reading
10
High-Performance Liquid Chromatography for Analysis of Herbal Drugs
	HPLC Analysis for Evaluation of Herbal Drugs
	Preparation of Sample for HPLC
	Normal-Phase Chromatography
		Mechanism of Retention
		Stationary Phases for Normal-Phase Chromatography
		Mobile Phases for Normal-Phase Chromatography
	Reversed-Phase Chromatography
		Mechanism of Retention
		Stationary Phases for Reversed-Phase Chromatography
		Mobile Phases for Reversed-Phase Chromatography
	Factors Affecting HPLC
		Capacity Factor
		Resolution
		Selectivity
		Efficiency
	Instrumental Features in HPLC Analysis
		Solvent Delivery Systems
		Pumps
			Syringe Pumps
			Reciprocating-Piston Pumps
			Isocratic Pumping Systems
			Gradient Pumping Systems
		Columns
			Stationary Phases
			Pore Size
			Particle Size
			Internal Diameter of Column
			Column Length
		Detector
			UV–Visible Absorbance Detector
			Fluorescence Detectors
			Electrochemical Detector
			Conductivity Detector
			Refractive Index Detector
			Evaporative Light Scattering Detector
			Corona Charged Aerosol Detection
			Detectors Hyphenated with HPLC
		Properties of Detectors
			Noise
			Sensitivity
			Accuracy
	Validation of RP-HPLC Methods
		Accuracy
		Precision
		Specificity
		Linearity
		Range
		Quantitation Limit
		Robustness
	Chemometrics and Principal Component Analysis
	Case Studies for HPLC in the Analysis of some Phytochemical Groups and Herbal Products
		Alkaloids
		Phenolics
		Coumarin
		Flavonoids
		Lignans
		Terpenes (Mono, Sesquiterpenes, Diterpenes, and Triterpenes)
		Triterpenes
		Carotenoids
		Some Case Studies With HPLC Analysis of Few Herbs and Their Components
			Estimation of Aristolochic Acid in Medicinal Plants
			Estimation of Ginsenosides in Panax ginseng
			RP-HPLC for Fingerprint Quality Evaluation of Hypericum japonicum
			Estimation of Mangiferin in Mangifera indica
			Simultaneous Estimation of Ten Compounds in Tibetan Herb, Pterocephalus hookeri by UPLC
			Determination of Baicalin and Puerarin by RP-HPLC
			Simultaneous Estimation of Hydrastine and Berberine
			Estimation of Boswellic Acid From Market Formulations Containing Boswellia serrata Extract
			RP-HPLC Analysis of the Phenolic Compounds in Menta pulegium
			Validated RP-HPLC Method for the Standardization of Phyllanthus niruri (Herb and Commercial Extracts) Using Corilagin
	HPLC Standardization of Some Medicinal Plant With Their Chromatograms
		Tinospora cordifolia HPLC Quantification of Tinosporaside
		Mucuna pruriens
		Swertia chirata
		Trigonella foenum-graecum
		Syzygium aromaticum
		Centella asiatica
		Piper longum
	References
	Further Reading
11
LC–MS: A Rapid Technique for Understanding the Plant Metabolite Analysis
	Basic Theory and Principle
	Ionization Sources
		Electrospray Ionization (ESI)
		Atmospheric Pressure Chemical Ionization (APCI)
		Atmospheric Pressure Photoionization (APPI)
		Thermospray
		Matrix-Assisted Laser Desorption/Ionization (MALDI)
	Mass Analyzer
		Quadrupole Analyzer
		Time-of-Flight (TOF)
		Ion Trap
		FT-ICR
	Detectors
		Photographic Plates
		Faraday Cup
		Electron Multiplier
		Scintillation Detectors
	Data Handling, Libraries, and Database
	Statistical Methods for Analysis of LC–MS Data
		Principal Component Analysis (PCA)
		Cluster Analysis
		Soft-Independent Model of Class Analogy (SIMCA)
		Partial Least-Squares Discriminant Analysis (PLS–DA)
		Heat Map
		Quantitative Trait Loci (QTL)
	LC–MS in Metabolite Profiling and Dereplication of Herbal Drugs
	LC–MS in Quality Control and the Identification and Quantification of Adulterants
	Chemotaxonomic and Phylogenetic Characterization of Medicinal Plants
	LC–MS in Target Compound Analysis
	LC–MS in Pharmacokinetics and Bioequivalence Study of Herbal Drugs
	References
	Further Reading
12
Chemoprofiling and Marker Analysis for Quality Evaluation of Herbal Drugs
	Importance of Chemoprofiling and Marker Analysis
	Classification of Markers Present in Herbal Drugs
		Active Principles
		Active Marker(s)
		Analytical Markers
		Negative Markers
	Botanical Reference Standard (BRS) and Phytochemical Reference Standard (PRS)
	DNA-Based Marker Analysis
	DNA Barcoding for Identification of Herbs
		Development Steps for Building a DNA Barcoding of Herbal Materials
			DNA Extraction
			PCR Amplification and Sequencing
			Sequence Assembly
			Species Identification
		Significance of DNA Barcoding
		Restriction Fragment Length Polymorphism (RFLP)
		Simple Sequence Repeats (SSRs)
		Random Amplification of Polymorphic Deoxyribonucleic Acid (RAPD)
		Amplified Fragment Length Polymorphism (AFLP)
		Intersimple Sequence Repeat (ISSR)
		Sequence Characterized Amplified Region (SCAR)
	Chemometrics in Marker Analysis
		Tools for Preprocessing Knowledge
			Normalization
			Local Least Square (LLS) Technique
		Tools for Extracting Chemical Data
			SCC (Spectral Correlative Chromatography)
			Information Theory (IT)
		Tools for Resolution of Mixtures
			HELP (Heuristic Evolving Latent Projections)
			OPA (Orthogonal Projection Analysis)
		Tools for Displaying Data
			PCA (Principal Component Analysis)
	Chemometric Analysis in Phytochemical Characterization
	References
	Further Reading
13
Bioassay-Guided Isolation and Evaluation of Herbal Drugs
	Bioassay and Its Importance in the Evaluation of Herbal Drugs
	In Vivo-Based Bioassay
		Brine Shrimp Model
		Zebrafish Model
		Drosophila Model
		Animal Model
	Bioassay in Isolated Organs
	Cell-Based Bioassay
		Cell Viability Assays
		Cell Proliferation Assays
		Cytotoxicity Assays
		Second Messenger Assay
		Reporter Gene Assay
	Enzyme-Based Bioassay
	Receptor-Based Bioassay
	High-Throughput Screening
	Dereplication and Other Approaches for Bioassay-Guided Isolation
		Separation Methods
			Column Chromatography
			Vacuum Liquid Chromatography
			Flash Column Chromatography
			Solid Phase Extraction
			Thin-Layer Chromatography and High-Performance Thin-Layer Chromatography
			High-Performance Liquid Chromatography
			Ultra-High-Performance Liquid Chromatography
			Gas Chromatography
			Capillary Electrophoresis
		Hyphenated Techniques
			Liquid Chromatography–Mass Spectrometry
			Gas Chromatography–Mass spectrometry
			Liquid Chromatography–Nuclear Magnetic Resonance
		X-Ray Crystallography
		“Omics”-Guided Approach
			Genomics
			Proteomics
			Metabolomics
		In Silico Dereplication
	References
	Further Reading
14
Therapeutic Evaluation of Herbs With Enzyme Inhibition Studies
	Importance of Enzyme Inhibition Studies in the Evaluation of Herbal Medicine
	Different Strategies for Enzyme Inhibition Studies
	Inhibition Study of Acetyl Cholinesterase and Butyl Cholisterase (AChE/BChE)
		Importance of Acetyl Cholinesterase and Butyl Cholisterase (AChE/BChE)
		Assay Procedure for AChE/BChE Inhibition
		Natural Products as AChE/BChE Inhibitors
	Inhibition Study of Angiotensin Converting Enzyme (ACE)
		Importance of ACE
		Assay Procedure for ACE Inhibition
		Natural Products as ACE Inhibitors
	Inhibition Study of Pancreatic Lipase
		Importance of Pancreatic Lipase
		Assay Procedure for Lipase Inhibition
		Natural Products as Pancreatic Lipase Inhibitors
	Inhibition Study of HMG-COA Reductase
		Importance of HMG-COA Reductase
		Assay Procedure for HMGR Inhibition
		Natural Products as HMG-COA Inhibitors
	Inhibition Study of α-Glucosidase
		Importance of α-Glucosidase
		Assay Procedure for α-Glucosidase Inhibition
		Natural Products as α-Glucosidase Inhibitors
	Inhibition Study for Rhokinase
		Importance of Rhokinase
		Assay Procedure for ROCK Inhibition
		Natural Products as ROCK Inhibitors
	Inhibition Study of Carbonic Anhydrase
		Importance of Carbonic Anhydrase
		Assay Procedure for Carbonic Anhydrase Inhibition
	Inhibition Study of Hyaluronidase
		Importance of Hyaluronidase
		Natural Products as Hyaluronidase Inhibitors
		Assay Procedure for Hyaluronidase Inhibition
	Inhibition Study of Matrix Metalloproteinase (MMP)
		Importance of Matrix Metalloproteinase Enzyme
		Assay Procedure for MMP Inhibition
		Natural Products as MMP Inhibitors
	Inhibition Study of Tyrosinase
		Importance of Tyrosinase
		Assay Procedure for Tyrosinase Inhibition
		Natural Products as Tyrosinase Inhibitors
	Inhibition Study of Dipeptidyl Peptidase-IV
		Importance of Dipeptidyl Peptidase-IV
		Assay Methods for DPP-IV Inhibition
			Fluorogenic Substrate Assay
			Chromogenic Substrate Assay
			DPP Glo Protease Assay
		Natural Products as DPP-IV Inhibitors
	Inhibition Study of α-Amylase
		Importance of α-Amylase
		Assay Procedure for α-Amylase Inhibition
			DNS (3,5-Dinitro Salicylic Acid) Method
			The Fuwa Method (Starch–Iodine Method):
		Natural Products as Amylase Inhibitors
	References
	Further Reading
15
Evaluation of Herbal Drugs for Antimicrobial and Parasiticidal Effects
	Herbs as Antimicrobials
	Methods for Antimicrobial Evaluation
		Diffusion Method
			Agar Disk-Diffusion Method
			Kirby–Bauer Method
			Antimicrobial Gradient Method (E-Test)
		Dilution Method
			Broth Dilution Method
			Agar Dilution Method
		Bioautographic Methods
			Agar Diffusion
			Direct Bioautography
			Agar Overlay Bioassay
		Time-Kill Test
		ATP Bioluminescence Assay
		Flow Cytofluorometric Technique
	Antibacterial Screening Method
	Herbs With Antimicrobial Potentials
	Lead Phytoconstituents With Antimicrobial Activity
		Plant-Derived Antimicrobials
		Antimicrobials From Marine Sources
	Evaluation of Phytopharmaceuticals Having Antifungal Activity
		Screening Methods for Antifungal Agents
		Sensitivity Test
		Turbidity Method
		Spore Germination Method
		Disk-Diffusion Method
		High-Throughput Method for Antifungal Activity
	Evaluation of the Antimycobacterial Activity of Herbs
	Evaluation of Antimalarial Activity
		Selection of Blood Schizontocides for In Vivo Models
		Peters’ Test
		The Rane Test
		In Vitro Models for the Selection of Blood Schizontocides
		Toxicity to Macrophages
		In Vitro Antimalarial Assays
	Antigiardial Effect of Herbs and Its Evaluation
		In Vitro Activity
		In Vivo Screening
			Specific Immune Response
			Nonspecific Immune Response
	Amoebicidal Activity of Herbs and Its Evaluation
		In Vivo Models for Amoebiasis
		In Vitro Models for Amebiasis
			Materials and Medium for In Vitro Amoebicidal Test
			Maintenance of Cultures
			Amoeba Cell Count
			Cryopreservation of Amoebae
			Amoebicidal Test Protocol
	Antimicrobial Resistance
		Mechanisms for Antimicrobial Resistance
		Test Methods Used for Antimicrobial Resistance
		Herbs Having Anti-MRSA Potential
	References
	Further Reading
16
Antiviral Evaluation of Herbal Drugs
	Antiviral Potentials of Herbs
	Phytoconstituents Having Antiviral Potentials
	In Vitro Perspectives for Antiviral Evaluation
		MTT and Neutral Red (NR) Assays
		WST-1 Cell Proliferating Reagent Assay
	Factors Influencing Antiviral Evaluation
	Host Cells, Organisms, and Culture Media
	Methods for Antiviral Evaluation of Herbal Drugs
		Preparation of Samples for Antiviral Testing
		Extracellular Viricidal Evaluation Procedure
		End Point Titration Technique (EPTT)
		Effect of Test Compounds on Virus CPE Inhibition
		Virus Yield Assay
		MTT Assay
		Plaque Reduction Assay
		Inhibition of Virus Hemagglutination Activity (HA)
		Immunofluorescence Assay
		Enzyme-Linked Immunosorbent Assay
		Virus Inactivation Assay
		Attachment Assay
		Penetration Assay
		Virus Adsorption Assay
	Virus-Specific Antiviral Assays
		Human Immunodeficiency Virus (HIV)
		Hepatitis B Virus
	Cytotoxicity Studies
		Cytotoxic Study by Trypan Blue Dye Exclusion Technique
		Determination of Cell Metabolic Function by Protein Estimation
		Determination of Mitochondrial Synthesis by MTT Assay
		Brine Shrimp Bioassay
			Lethal Concentration Determination
	References
	Further Reading
17
Plant Metabolomics and Quality Evaluation of Herbal Drugs
	Metabolomics: A New “Omics” Platform Technology
	Targeted and Untargeted Metabolomics
		LC–MS-Based Techniques
		GC-MS-Based Techniques
		LC-NMR-Based Techniques
	Metabolomics in Systems Biology
	Application of Metabolomics in Herbal Medicine
	Qualitative and Quantitative Study of Herbal Medicine
	In Vitro, In Vivo, and Clinical Studies of Herbal Medicine Through Metabolomics
	Statistical Analysis of Metabolomics Data
	References
	Further Reading
18
Safety-Related Quality Issues for the Development of Herbal Drugs
	Safety Concerns for Herbs
	Quality-Related Safety Issues of Herbal Medicines
	Toxicity Evaluation of Herbal Medicine
		Methods for Toxicological Evaluation of Herbs
			Preclinical Toxicity Test
			Toxicity Evaluation by Cell-Based Cytotoxicity Tests
			Determination of Toxicokinetic Properties of Herbs
			Tools for Toxicogenomic Screening
			High-Throughput DNA Sequencing
			Using In Vivo Animal Models
				Determination of Systemic Acute Toxicity
				Determination of Subacute or Subchronic Toxicity
			Determination of Carcinogenicity or Chronic Toxicity
			Evaluation of Specific Toxicity
			Tests for Evaluation of Local Toxicity
				Determination of Neurotoxicity
			Determination of Genotoxicity
			Determination of Reproductive Toxicity Development
	Synergistic Effects of Herbal Medicines
	Herb–Drug Interactions
	Validation of Herbal Therapies
	Effects of Herbs on CYP450 Enzyme Inhibition and Induction
		Cytochrome P450: Its Role in Metabolism
		Induction of CYP P450-Mediated Drug Metabolism
		Inhibition of Cytochrome P450-Mediated Drug Metabolism
			Reversible Inhibition
			Noncompetitive Inhibition
				Uncompetitive Inhibition
			Mixed Inhibition
			Time-Dependent Inhibition
			Quasiirreversible Inhibition
			Irreversible Inhibition
	Pharmacovigilance for Herbal Medicines
		Phytovigilance
		Challenges in Pharmacovigilance: How Safe Are Your Herbs?
		Steps to Commence Pharmacovigilance for Herbal Products
		Postmarketing Surveillance
	Adverse Drug Reactions in Herbal Medicine
		Assessment of Severity and Seriousness of ADRs
	References
	Further Reading
19
Quality Assurance of Herbal Drugs and Stability Testing
	Quality Assurance and Stability Testing of Herbal Drugs
		Quality Assurance Measures
			Good Agricultural and Collection Practice (GACP) and Species Authentication Using Classical Systematics
			Genomic Profiling and DNA Barcoding in Species Authentication
			Plant Authentication Based on Macroscopic and Microscopic Features
			Phytochemical Profiling of Plant Parts as a Tool for Identification and Characterization
			Guidelines for Good Plant Authentication and Identification Practice (GPAIP)
			Assurance of the “Purity” of Botanical Raw Materials: Impurity Profiling
		Challenges in Quality Assurance of Herbal Medicine
	Use of Indicative Substances
		Indicative Substances for Identification
		Indicative Substances for Content Determination
		Indicative Substances for Stability Testing
			Analytical Methods
				Ambient Ionization Techniques
				Desorption Electrospray Ionization
				Direct Analysis in Real Time
	GMP and HACCP in Traditional Systems of Medicine
		Conduct a Hazard Analysis
		Determine CPPs
		Establish Critical Limits
		Establish Monitoring Procedures
		Establish Corrective Actions
		Establish Record keeping and Documentation Procedures
		Establish Verification Procedures
	Physical Quality Assurance
	Quality Assurance by Cultivation and Breeding
	Stabilization and Stability
	Stabilization Methods
		Guidelines for Stability Testing
		Real-Time Stability Testing
		Accelerated Stability Testing
		Retained Sample Stability Testing
		Cyclic Temperature Stress Testing
		Stability Testing With the Help of Drying
	Factors Affecting Stability for Herbal Formulations
		Effect of pH
		Concentration
		Light
		Temperature
		Relative Humidity (RH)
		Water Activity (AW)
		Atmosphere
	Validation of Analytical Procedures
		Identity Testing
		Specificity
		Linearity
		Precision
		Trueness
		Detection and Determination Limits
		Sensitivity
		Ruggedness of the Method
	References
	Further Reading
20
Phyto-Pharmaceuticals, Nutraceuticals and Their Evaluation
	Nutraceutical and Dietary Supplements From Botanicals
	Plant-Made Pharmaceuticals
	Health Benefits of Nutraceuticals and Functional Foods
	Developmental Factors of Botanicals as Nutraceuticals
	Classification of Nutraceuticals
		Bioactive Compounds and Phytochemicals
			Curcumin
			Anthocyanins
	Nutraceutical and Dietary Supplements for Obesity Management
		Berries
		Blackberry (Family: Rosaceae)
		Indian Gooseberry (Family: Phyllanthaceae)
		Rubus idaeus (Family: Rosaceae)
		Solanum lycopersicum (Family: Solanaceae)
		Capsicum
	Nutraceutical and Dietary Supplements for Geriatrics
		Antioxidants
			Plant Polyphenols and Catechins
			Carotenoids: Xanthophyll, Zeaxanthin, and Carotenoid
		Plant Sterols (Phytosterols)
		Vitamin-B
		Calcium and Vitamin D
		Omega-3-Fatty Acid
		Glucosamine, Chondroitin, Collagen
		Dietary Fiber
		Prebiotics and Probiotics
		Whey Protein
		Zinc
		Rasayana Therapy in Ayurveda
	Nutraceuticals Used for Cardiovascular Diseases
		Terminalia arjuna
		Garlic
		Tinospora cordifolia
		Emblica officinalis
	Validation of Botanicals as Nutraceuticals
		Safety of Nutraceuticals
		Regulatory Aspects of Phyto-Pharmaceuticals, Dietary Supplements, and Nutraceuticals
	References
	Further Reading
21
Regulatory Harmonization and Good Quality Practices for the Development of Herbal Medicine
	Regulatory Harmonization in Herbal Drug Development
	Regulatory Status of Herbal Drugs in India
		Ayurvedic, Siddha, and Unani Drugs Technical Advisory Board (ASUDTAB)
		Ayurvedic, Siddha, and Unani Drugs Consultative Committee (ASUDCC)
		Misbranded Drugs
		Adulterated Drugs
		Spurious Drugs
		Rules of Manufacture for Ayurvedic, Siddha, and Unani (ASU) Drugs
		Licensing Authorities
	Good Harvesting Practices
		Collection of Plant Materials
		Problems Encountered in the Collection of Medicinal Plants
		Good Collection Practices
	Good Agricultural Practices
		SOPs for the Controlled Cultivation of Medicinal Plants
		Principles and Guidelines for GAP
			Seeds and Propagation Material
			Cultivation
		Harvesting of Cultivated Medicinal Plants
		Collection of Medicinal Plants in the Wild
		Primary Processing of Herbal Products
		Packaging of Collected Materials
		Storage and Transport
		Equipment and Machinery
		Requirements for the Staff
		Requirements for Buildings and Related Items
	Good Laboratory Practices
		Principles of GLP
	Good Manufacturing Practice
	Good Clinical Practice
		GCP for Indian Systems of Medicine: Ayurveda, Siddha, and Unani
	Quality Reporting of Herbal Clinical Trials
	Intellectual Property Rights and Patent Issues With Herbal Medicines
		Patents of Herbal Drugs
		Patentability Requirements
		Inventive Step and Nonobviousness
	International Regulatory Status of Herbal Medicine
		Australia
		Canada
		China
		European Union
	References
	Further Reading
Index
	A
	B
	C
	D
	E
	F
	G
	H
	I
	J
	K
	L
	M
	N
	O
	P
	Q
	R
	S
	T
	U
	V
	W
	X
	Y
	Z
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