Unit 5

INTRODUCTION:
Recently, herbal medicinal products (HMP) have gained importance and are
extensively used in the prevention and treatment of various diseases. A
commercial herbal medicinal product should comply with the regulatory
requirements of quality, safety, and efficacy. Currently, the standards and
regulations of herbal medicinal products vary from country to country, which
poses a challenge to the manufacturing companies to place a standardized
herbal product in the international market.
Regulations for Herbal Medicines:
In India, the Department of Ayurveda, Yoga and Naturopathy, Unani, Siddha,
and Homoeopathy (AYUSH) is the regulatory authority that controls product
licensing, composition, formulation and ensures manufacturing of products,
labeling, packing, and quality as per Schedule "T" for good manufacturing
practice (GMP) and monitors export of Ayurveda, Siddha and Unani products.
Herbal products are licensed under Ayurveda, Siddha, and Unani drugs, as
patent or proprietary medicines.
Herbal medicinal product development flow

Different regulations for herbal medicine are

• Good Agricultural and Collection Practice (GACP)
• Good Manufacturing Practice (GMP)
• Good laboratory Practice (GLP
 • Good Clinical Practice (GCP)

WHO GUIDELINES ON SAFETY MONITORING OF HERBAL

MEDICINES IN PHARMACOVIGILANCE SYSTEMS
WHO guidelines provide practical technical guidance for monitoring the safety
of herbal medicines within pharmacovigilance systems. The global consumption
of herbal medicines is enormous so that in terms of population exposure alone,
it is essential to identify the risks associated with their use. The safety of herbal
medicines is an important public health issue.
Herbal medicines are frequently used in conjunction with other medicines and it
is essential to understand the consequences of such combined use and monitor
whether any adverse effects are arising. This can be achieved most readily
within existing pharmacovigilance systems.
Problems:
Among consumers, there is a widespread misconception that "natural" always
means "safe" and a common belief that remedies from a natural origin are
harmless and carry no risk. However, some medicinal plants are inherently
toxic. Adverse effects may arise from the mistaken use of the wrong species of
medicinal plants, incorrect dosing, and errors in the use of herbal medicines
both by healthcare providers and consumers. Interact ions with other medicines
and use of products contaminated with hazardous substances such as; toxic
metals, pathogenic micro-organisms, and agrochemical residues can also cause
adverse effects.
The objectives of these guidelines are
1. To support Member States, in the context of the WHO International
Drug Monitoring Programme.
2. To provide practical technical guidance for monitoring the safety of
herbal medicines within pharmacovigilance systems.
3. To include herbal medicines in National drug safety monitoring
systems.
4. To provide standard definitions of terms relating to pharmacovigilance
and safety monitoring of herbal medicines.
5. To promote the safe and proper use of herbal medicines.
The WHO guidelines were developed with the view that, within current
pharmacovigilance systems, monitoring of the safety of medicines should be
enhanced and broadened in ways that will allow the successful monitoring of
herbal medicines. It is not the intention to suggest that different systems should
be instituted for this purpose.
However, because of the unique characteristics of the provision and use of
herbal medicines, several technical issues need to be addressed if adequate and
effective monitoring is to be introduced. The guidelines, therefore, identify the
particular challenges posed in monitoring the safety of herbal medicines
effectively and propose approaches for overcoming them. Special attention is
also given to the reporting system for adverse reactions to herbal medicines and
to the analysis of the causes of the reported adverse reactions.
Currently, the majority of adverse events related to the use of herbal products
and herbal medicines that are reported are caused by poor product quality or
improper use of herbal products. Member States are, therefore, encouraged to
strengthen national regulation, registration quality assurance, and control of
herbal medicines. Also, national health authorities should give greater attention
to consumer education and to qualified practice in the provision of herbal
medicines.
COMPARISON OF VARIOUS HERBAL PHARMACOPOEIAS

Herbal pharmacopoeias:
Herbal pharmacopoeias are official publications or
reference books containing a systematic list of
medicinal plants, descriptions, and standards for their
identification, quality, and purity. It includes the
preparation of quality medicines published by the
authority of a Government and represents qualitative
and therapeutic monographs on botanicals These
pharmacopoeias are crucial for the standardization and
regulation of herbal products, ensuring their safety,
efficacy, and quality. Countries and regions have herbal
pharmacopeias, which may vary in scope, detail, and
regulatory authority.
Criteria for Herbal Pharmacopoeia:
1. It is an important reference for all individuals and
organizations working within pharmaceutical
research and development, manufacture and testing
around the globe
2. Herbal Pharmacopoeia is a compiled document
which:

(a) Defines a botanical drug.

(b) Provides information for its proper identification.
(c) Contains the basic description including;
nomenclature, plant parts used, active constituents,
range of application, contraindications and side
effects, incompatibilities with other medications,
dosage, use and action of the herb.
3. The modern-day pharmacopoeias are strictly for
standards of medicines and every developed
country has a pharmacopoeia for their medicaments
of synthetic as well as natural origin.

Types of Pharmacopoeias :
1. European Pharmacopoeia (Ph. Eur.)
2. United States Pharmacopoeia (USP)
3. American Herbal Pharmacopoeia (AHP)
4. Japanese Pharmacopoeia (JP)
5. African Herbal Pharmacopoeia (AfrHP)
6. British Herbal Pharmacopoeia (BHP)
7. Korean Herbal Pharmacopoeia (KHP)
8. Pharmacopoeia of the People’s Republic of
China (ChP)
9. The Turkish Pharmacopoeia (TP)
 10. Pharmacopoeia Argentina
11. Brazilian Pharmacopoeia
12. Indian Herbal Pharmacopoeia
a. Ayurvedic Pharmacopoeia of India (API)
b. Siddha Pharmacopoeia of India

European Pharmacopoeia (Ph. Eur.)

Region: Europe
Authority: European Directorate for the Quality of
Medicines & HealthCare (EDQM)
Content: Includes monographs on herbal drugs,
extracts, and essential oils used in European herbal
medicine. Standards cover identification, purity, and
assay methods.
Features: High standards for analytical methods, focus
on harmonization across European countries.
United States Pharmacopoeia (USP)
Region: United States
Authority: United States Pharmacopeial Convention
Content: Monographs on herbal supplements, including
identity, strength, quality, and purity standards.
Features: Emphasis on safety and efficacy, rigorous
testing protocols, and widely recognized standards.
American Herbal Pharmacopoeia (AHP)
The AHP develops qualitative and therapeutic
monographs on botanicals including; many of the
Ayurvedic, Chinese and Western herbs most frequently
used in the United States. These monographs represent
the most comprehensive and critically reviewed body of
information on herbal medicines in the English
language and serve as a primary reference for
academicians, health care providers, manufacturers and
regulators.

Japanese Pharmacopoeia (JP)

Region: Japan
Authority: Ministry of Health, Labour and Welfare
Content: Monographs on Kampo medicines (traditional
Japanese herbal medicines) and Western herbal
medicines used in Japan. Standards include botanical
and chemical identification.
Features: Integration of traditional and modern herbal
medicines, detailed quality control measures.
African Herbal Pharmacopoeia (AfrHP)
The AfrHP provides comprehensive, up to date
botanical, commercial and phytochemical information
on over 50 of the most important African medicinal
plants. The technical data were made on plant samples
sourced from across the continent. These monographs
were prepared by leading African scientists and have
been reviewed by international experts. Additional data
includes; micro-morphology of the plant material,
distribution maps and TLC chromatograms. These data
are crucial for producers, collectors and traders in
medicinal plants and extracts as well as researchers,
manufacturers and practitioners. The scope, quality and
standard of these herbal monographs are comparable to
those prepared in Europe, North America and Asia.
3. British Herbal Pharmacopoeia (BHP):
British Herbal Pharmacopoeias:

The BHP provides quality standards for 169 herbal raw

materials. European Pharmacopoeia Commission
(Council of Europe) has led to the introduction of many
more herbal monographs in the European
Pharmacopoeia. The BHP monograph includes a brief
description of the form of the material, its main
geographical sources, macroscopical and microscopical
descriptions of the herbal material, prepared by eminent
pharmacognosists, together with odour and taste
descriptions. It also includes; comparative identification
and qualitative phytochemical analysis. However, the
BHP remains a very useful aid to quality assurance,
particularly for herbs not featured in official
pharmacopoeias.

Korean Herbal Pharmacopoeia (KHP) Herbalh

The recent KHP contains a total of 2,462 official
monographs (Part I and Part II). Part I includes;
frequently used drugs and their primary preparations
and Part II includes; crude drugs, biological drugs,
radioactive drugs, combined preparations, additives for
preparations and quasi-drugs. The Pharmacopoeia
comprises the following items, in order, general notices,
general requirements for preparations, monographs,
general tests, processes and apparatus and general
information followed by the index.
Chinese Pharmacopoeia (ChP) Pharmacopoeia of
Region: China
Authority: Chinese Pharmacopoeia Commission
Content: Includes monographs on traditional Chinese
medicine (TCM) herbs, formulations, and decoction
pieces. It provides standards for identification, purity,
and potency.
Features: Extensive focus on traditional Chinese
medicinal herbs, detailed descriptions, and advanced
analytical techniques for quality control.
The Ministry of Health published the Republic of
China’s first pharmacopoeia as Chinese Pharmacopoeia
in 1953. Under the umbrella of the China Food and
Drug Administration (CFDA), the Chinese
Pharmacopoeia Commission (ChPC) and its
stakeholders (drug control institutions, research
institutions, universities and drug manufacturers)
prepared the Chinese Pharmacopoeia in line with the
basic principles, goals and requirements of the era. The
Chinese Pharmacopoeia includes; monographs of
vegetables, oil, fats and extracts, medicinal materials
and the prepared slices of Chinese crude drugs, single
item preparations, chemical drugs, antibiotics,
biochemical and radioactive drugs, pharmaceutical
excipients and biologicals. General chapters include;
general requirements of preparations, testing methods,
standard substances, reagents and the guidelines

6. The Turkish Pharmacopoeia (TP):

Turkish Pharmacopoeia includes European
Pharmacopoeia monographs as well as national
monographs. It has a section containing herbal,
biological or biotechnological products and finished
products. They are first published in the Turkish
Pharmacopoeia Journal and then in the Turkish
Pharmacopoeia and become official. In the latest
version, the number of national monographs has
reached 32 and most of them are monographs of
medicinal and aromatic herbal ingredients.

Pharmacopoeia Argentina is published by The

National Administration of Medicines Food and
Medical Technology and it is in the Spanish language.
Pharmacopoeia Argentina provides a specification for
biological, herbal products, medical products, blood
products, radiopharmaceutical products, for vaccines
and includes general monographs and supplementary
texts.
Brazilian Pharmacopoeia is published by
Pharmacopoeia Coordination Brazilian Health
Surveillance Agency in three different languages;
Portuguese, English and Spanish. Apart from Brazilian
Pharmacopoeia, Herbal Medicines National Formulary
(Portuguese/ Spanish) and Herbal Medicines Memento
(Portuguese) are also used in Brazil. Brazilian
Pharmacopoeia provides specifications via monographs
for APIs and excipients, for dosage forms, for
biological, for herbal products and includes
monographs for medical devices.
Indian Pharmacopoeia (IP)

Region: India
Authority: Indian Pharmacopoeia Commission
Content: Contains monographs on Ayurvedic, Siddha,
and Unani medicinal plants and formulations. Standards
include botanical, chemical, and pharmacological
parameters.
Features: Emphasis on traditional Indian medicine
systems, comprehensive guidelines for quality and
safety assessment.rbal Pharmacopoeia (IP):
The Indian Herbal Pharmacopoeia has 40 monographs
published by the Indian Drugs Manufacturers
Association (IDMA) in collaboration with Regional
Research Laboratory (RRL), Jammu. Index of
pharmacopoeias published by the World Health
Association, Geneva has a comprehensive list of
pharmacopoeias of different countries.

Ayurvedic Pharmacopoeia of India (API) has

monographs of 600 plants/animals/minerals and derived
drugs (Part 1. Volume 1 to 8). Monographs of 152
compound Ayurvedic formulations are published in
Ayurvedic Pharmacopoeia of India (Part II, Volume 1
to 3).

Other than APL Siddha Pharmacopoeia of India

(Volume 1 and 2), Homeopathic Pharmacopoeia of
India (Volume 1 to 6) and Unani Pharmacopoeia of
India (Volume 1 to 6) are the standard books available
for Indian herbs.

The innovative work, Quality standards of Indian

Medicinal Plants by the Indian Council of Medical
Research (ICMR) are a non-pharmacopoeial
masterpiece for reference on standards of Indian herbs.
Monographs on 449 plants (Volume 1 to 13) included
in the book would help in improving monographs in
API and other Pharmacopoeias of Indian origin. The
Pharmacopoeia Commission for Indian Medicine
(PCIM) under the Ministry of AYUSH is fully
functional to improve the Pharmacopoeias for Indian
Herbs.
The first official Pharmacopoeia of India was published
in 1868. In 1946, the Government of India issued one
list known as “The Indian Pharmacopoeial list. The
committee under the chairmanship of Sir R. N. Chopra
along with the other nine members prepared The Indian
Pharmacopoeial list. It was prepared by the Department
of Health, Govt of India, Delhi in 1946. In 1948, the
Government of India appointed an Indian
Pharmacopoeia committee for preparing
“Pharmacopoeia of India. The tenure of this committee
was 5 years. Indian Pharmacopoeia committee under
the chairmanship of Dr B. N. Ghosh published the first
edition of IP in 1955. Supplement to this edition was
published in 1960.
The Second edition of IP was published in 1966 under
the chairmanship of Dr B. Mukherjee. Official titles of
monographs were given in English. Formulations of the
drugs were given immediately after the monograph of
drugs. 274 monographs from IP 1955 and their
supplements were deleted. Supplement to this edition
was published in 1975. In this, 126 new monographs
were included and 250 monographs were amended.
The Third edition of IP was published in 1985 with two
volumes and nine appendices, 261 new monographs
were added. Addendum I to IP was published in 1989
where 46 new monographs were added and 126
amended. Addendum II was published in 1991 where
62 new monographs were added and 110 amended.
The Fourth edition of IP was published in 1996 under
the chairmanship of Dr Nityanand. It covered 1149
monographs and 123 appendices. It included 294 new
monographs and 110 monographs were deleted.
Addendum I was made effective from 31 December
2000 where 42 new monographs were added.
Addendum I was made effective from 30 June 2003
where 19 new monographs were included.
The Fifth edition of IP was published in 2007 and an
addendum to this edition was published in 2008. IP
2007 was presented in three Volumes. Volume 1
contains general notices and general chapters. Volume
II and III contain general monographs on drug
substances, dosage forms and pharmaceutical aids.
The Sixth edition of IP was published in 2010 by the
Indian Pharmacopoeia Commission (IPC). The Indian
Pharmacopoeia 2010 was presented in three volumes.
Volume I contains the notices, preface, the structure of
the IPC, acknowledgements, introduction and the
general chapters. Volume II contains the general notice,
general monographs on dosage forms and monographs
on drug substances and pharmaceutical aids (A to M).
Volume III contains monographs on drug substances,
dosage forms and pharmaceutical aids (N to Z). It also
includes monographs on vaccines and immunosera for
human use, herbs and herbal products, blood and blood-
related products, biotechnology products and veterinary
products. The number of monographs of excipients,
anticancer drugs, herbal products and antiretroviral
drugs has been increased in this edition. A chapter on
Nuclear magnetic resonance spectroscopy (NMR) was
also incorporated in Appendices.
The Seventh Edition of the Indian Pharmacopoeia (IP
2014) was published by the Indian Pharmacopoeia
Commission (IPC) on behalf of the Government of
India, Ministry of Health and Family Welfare. The
Indian Pharmacopoeia 2014 was presented in four
volumes. The scope of Pharmacopoeia has been
extended to include; additional anticancer drugs and
antiretroviral drugs and formulations, products of
biotechnology, indigenous herbs, herbal products and
veterinary vaccines. The IP 2014 included 2550
monographs of drugs out of which 577 are new
monographs consisting of APIs, excipients, dosage
forms and herbal products, etc.
The Eighth Edition of the Indian Pharmacopoeia (IP
2018) was published by the Indian Pharmacopoeia
Commission (IPC) on behalf of the Ministry of Health
and Family Welfare, Government of India. IP 2018 was
brought out in four volumes incorporating 220 new
monographs including; chemical monographs (170),
herbal monographs (15), blood and blood-related
products (10), vaccines and immunosera for human use
monographs (02), radiopharmaceutical monographs
(03), biotechnology-derived therapeutic products (06),
veterinary monographs (14), 366 revised monographs
and 7 monographs have been deleted.

Comparison of Various Herbal Pharmacopoeias

 Chinese Indian European United States Japanese
Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopoeia
Feature (ChP) (IP) (Ph. Eur.) (USP) (JP)
Extensive TCM Ayurvedic, Siddha, European herbal Herbal Kampo and
herbs and Unani herbs and drugs, extracts, supplements, Western herbal
Scope formulations formulations essential oils dietary ingredients medicines
Chinese Indian United States Ministry of Health,
Pharmacopoeia Pharmacopoeia Pharmacopeial Labour and
Authority Commission Commission EDQM Convention Welfare
Botanical, Botanical, Botanical, Botanical, Botanical,
Identification microscopic, chemical, chemical, chemical, chemical,
Methods chemical pharmacological analytical analytical analytical
Detailed
monographs, Comprehensive High analytical Rigorous safety
Quality advanced guidelines for standards, and efficacy Detailed quality
Standards techniques quality, safety harmonization standards control measures
Advanced, Botanical,
Analytical including HPLC, chemical, HPLC, GC-MS, HPLC, GC-MS, HPLC, GC-MS,
Techniques GC-MS pharmacological spectrometry spectrometry spectrometry
Focus on
Traditional
Medicine High High Moderate Low High
International
Influence Growing Regional High High Regional

ROLE OF CHEMICAL AND BIOLOGICAL MARKERS IN THE

STANDARDIZATION OF HERBAL PRODUCTS
Importance of Standardization: Standardization of crude drugs means
confirmation of their identity and determination of their quality, purity, and
detection of the nature of adulterant by various parameters (pharmacognostic
parameters, physicochemical parameters, phytochemical parameters,
microbiological assay, and chromatographic analysis). Standardization of crude
drug is important, to assess the quality of the raw materials, estimate the amount
of active principal present in it, and achieve batch-to-batch consistency of the
finished products.
Chromatographic Fingerprinting:
Chromatographic fingerprinting is a chromatogram representing the
characteristics of the herbs. It is one of the most important methods for the
evaluation of herbal crude drugs. Chromatographic fingerprinting plays an
important role in the standardization of herbal crude drugs. They are used for
confirming the presence of different herbs in the polyherbal formulation as well
as in the quantification of marker compounds.
Marker Compounds:
Marker compounds are the pure and single isolated chemical constituents within
a crude drug that confirm the exact botanical identity of that starting material or
the crude drug. Marker compounds are of interest for quality control purposes
and they may or may not contribute to the therapeutic activity.
Role of markers in the standardization of herbal products:
As per WHO, there are three kinds of herbal medicinal products: raw plant
materials, processed plant materials, and medicinal herbal drugs. Herbal drugs
are finished and labelled products that contain active ingredients such as; aerial
or underground parts of plants or other plant materials or a combination thereof,
whether in the crude state or as plant preparations. The use of herbal medicines
has increased with the global trend of people returning to natural therapies.
Herbal medicinal products and dietary supplements that people take to improve
their health are sold as tablets, powders, capsules, teas, extracts, and fresh or
dried plants. Correct identification and quality assurance of the starting
materials is, therefore, an essential requirement to ensure the reproducible
quality of herbal medicine which contributes to its safety and efficacy.
Marker compounds may serve to calculate the amount of active components of
herbal drugs in the preparation or in the finished product. Markers are used at
various stages of the development and manufacturing of an herbal medicine
like; authentication and differentiation of species, collecting and harvesting,
quality evaluation, and stability assessment, diagnosis of intoxication, and
detection of lead compounds.
Types of Marker compounds:
1. Chemical Markers
2. Biochemical Markers
3. Molecular or DNA Markers
Definition of Chemical Markers
Chemical markers are specific compounds, often bioactive, found in herbal
products that can be quantitatively measured to assess the quality, potency, and
consistency of these products. They serve as reference points for the
identification and quantification of herbal ingredients.
Importance of Chemical Markers
 Quality Control: Chemical markers are used to confirm the identity of the
herbal product, ensuring that the correct plant species and plant part are
used. This helps in preventing adulteration and ensuring product purity.
 Consistency and Reproducibility: Herbal products can vary significantly
due to differences in growing conditions, harvest times, and processing
methods. By measuring chemical markers, manufacturers can ensure
batch-to-batch consistency.
 Efficacy: The therapeutic effects of herbal products are often attributed to
specific active compounds. Chemical markers help in ensuring that these
bioactive compounds are present in the required amounts to provide the
desired therapeutic effects.
 Safety: Monitoring chemical markers helps in detecting and quantifying
potentially harmful substances. This is important to ensure that the levels
of toxic compounds are within safe limits.
Selection of Chemical Markers
The selection of appropriate chemical markers is critical and is based on several
criteria:
 Biological Activity: Compounds that are known to contribute to the
therapeutic effects of the herb are preferred.
 Abundance: Markers should be present in significant amounts to be easily
detectable and quantifiable.
 Stability: Markers should be stable during processing and storage.
 Specificity: Markers should be unique to the plant species and not present
in adulterants.
 Techniques for Identifying and Quantifying Chemical Markers
Several analytical techniques are used to identify and quantify chemical
markers in herbal products:
 Chromatography: Techniques like High-Performance Liquid
Chromatography (HPLC) and Gas Chromatography (GC) are commonly
used to separate and quantify chemical markers.
 Mass Spectrometry (MS): Often coupled with chromatography, MS helps
in the precise identification and quantification of compounds.
 Spectroscopy: Methods like Nuclear Magnetic Resonance (NMR) and
Infrared (IR) spectroscopy are used for structural elucidation and
quantification.
 Bioassays: Sometimes, biological assays are used to assess the activity of
specific markers, particularly when the marker is a bioactive compound.
Regulatory Aspects
Regulatory agencies like the US FDA, EMA (European Medicines Agency), and
WHO have guidelines for the standardization of herbal products, often
recommending the use of chemical markers. These guidelines ensure that herbal
products meet specific standards for safety, efficacy, and quality.
Examples of Chemical Markers
 Ginsenosides in Ginseng: These saponins are used as markers to
standardize ginseng products.
 Curcumin in Turmeric: This compound is used to ensure the potency and
quality of turmeric supplements.
 Hypericin in St. John’s Wort: Used to standardize St. John’s Wort
preparations for their antidepressant properties.
Challenges and Considerations
 Complexity of Herbal Matrices: Herbal products contain numerous
compounds that can interfere with the detection and quantification of
markers.
 Variability: Natural variations due to environmental factors can affect the
concentration of chemical markers.
 Selection of Markers: Identifying the most relevant markers that
accurately reflect the quality and efficacy of the product can be
challenging.
Different Types of Chemical Markers
Applications of chemical markers:
(a) Identification of adulterants.
(b) Differentiation of herbal medicines with multiple sources.
(c) Determination of the best harvesting time.
(d) Confirmation of collection sites.
(e) Assessment of processing methods.
(f) Identification and quantitative determination of proprietary
products.
(g) To evaluate the stability of proprietary product quality over time
and determine the recommended shelf life.
(h) Diagnosis of herbal intoxication in which, toxic components
may be used as chemical markers in screening methods. e.g.:
Rapid diagnosis of acute hidden aconite poisoning in urine
samples by HPLC·MS and other techniques.
(i) Detection of lead compounds for new drug discovery. E.g.:
Gambogic acid is a potential lead compound for new anti-
cancer drugs.
Biochemical Markers:
Definition of Biological Markers
Biological markers are measurable indicators of some biological state or
condition. In the context of herbal products, they can include gene expression
levels, enzyme activities, protein expression patterns, or other physiological
changes that occur in response to the bioactive components of the herbal
product.
Importance of Biological Markers
Efficacy Assessment: Biological markers can directly reflect the biological
effects of herbal products. By measuring these markers, researchers can
evaluate the therapeutic efficacy of the product in a more relevant biological
context.
Mechanism of Action: They help elucidate the mechanisms by which the
herbal products exert their effects. Understanding these mechanisms can support
the rational use of herbal medicines and guide further research and
development.
• Safety Profiling: Biological markers can be used to monitor adverse
effects and toxicological responses, providing important safety
information about the herbal product.

• Quality Control: Just like chemical markers, biological markers can

help in maintaining batch-to-batch consistency and ensure that the
herbal product is biologically active.
Types of Biological Markers
• Pharmacodynamic Biomarkers: Indicators of the biological response
to the herbal product. For instance, changes in blood glucose levels
after administration of a herb used for diabetes management.
• Pharmacokinetic Biomarkers: Reflect the absorption, distribution,
metabolism, and excretion (ADME) of the active components of the
herbal product.
• Genomic Biomarkers: Changes in gene expression or genetic profiles
in response to the herbal product.
• Proteomic Biomarkers: Changes in protein expression or modification
patterns.
• Metabolomic Biomarkers: Alterations in metabolite profiles in
biological fluids or tissues.
Techniques for Identifying and Measuring Biological Markers
 • Genomics: Techniques such as DNA microarrays and RNA
sequencing to measure changes in gene expression.
• Proteomics: Methods like mass spectrometry and Western blotting to
identify and quantify proteins.
• Metabolomics: Techniques such as Nuclear Magnetic Resonance
(NMR) spectroscopy and liquid chromatography-mass spectrometry
(LC-MS) to profile metabolites.
• Cell-based Assays: Using cultured cells to assess biological activities
such as cell proliferation, apoptosis, or enzyme activity.
• Animal Models: In vivo studies to observe physiological and
pathological changes in response to the herbal product.
Applications of Biological Markers in Standardization
Identification of Active Components: By linking specific biological markers
with the presence of certain compounds, it’s possible to identify which
components are responsible for the therapeutic effects.
• Dose Optimization: Biological markers can help determine the most
effective and safe dosage of herbal products.
• Quality Assurance: Ensuring that the product consistently produces
the desired biological response, indicating a high level of quality
control.
• Clinical Validation: Biomarkers can be used in clinical trials to
provide evidence of efficacy and safety, supporting the acceptance and
standardization of herbal products.
Challenges and Considerations
• Complexity of Biological Systems: Biological responses can be
influenced by numerous factors, making it challenging to attribute
changes solely to the herbal product.
• Variability: Biological markers can vary widely between individuals
due to genetic, environmental, and lifestyle differences.
• Integration with Chemical Markers: Effective standardization often
requires a combination of both chemical and biological markers to
provide a comprehensive assessment.
• Regulatory Acceptance: The use of biological markers in
standardization requires validation and acceptance by regulatory
authorities, which can be a complex and rigorous process.
Molecular or Genetic markers or DNA markers:
DNA markers are the most widely used type of marker mainly due to their
abundance. DNA markers are unlimited in number and are not affected by
environmental factors, age and physiological conditions of the plant. ONA
can be extracted from fresh or dried plant tissue hence, the physical form of
the sample for assessment does not restrict detection.
Application of DNA markers:
• In Herbal drug technology: DNA-based molecular markers have
proved their utility in fields like; taxonomy, physiology,
embryology, and genetics.
• Genetic variation/genotyping: Random Amplified Polymorphic
DNA (RAPD) based molecular markers are useful in differentiating
different accessions of Andrographis paniculata, Azadirachta
indica, Allium schoenaprasum collected from different
geographical locations. Interspecies variation has been studied
using Restriction Fragment Length Polymorphism (RFLP) in
different genera such as; Glycyrrhiza, Curcuma, and Echinacea.
• Authentication of Medicinal plants: Rare and expensive medicinal
plant species are often adulterated or substituted by
morphologically similar, easily available, or less expensive species.
Sequence Characterized Amplified Region (SCAR), PCR, RAPD,
and RFLP are used for differentiating on of the plants and to detect
substitution or adulteration by other closely related species. E.g.:
Swertia chirata is frequently adulterated or substituted by
Andrographis paniculata.
• Medicinal plant breeding: Molecular markers have been used as a
tool to verify sexual and apomictic offspring of intraspecific
(means within the same species) cross in Hypericum perfaratum
(St. John's wort), a well-known diuretic and anthelmintic drug.
• Food and Nutraceuticals: Capsicum, maize, and soybean have been
successively categorized from non-GMO products.
• As a new pharmacognostic tool: DNA markers have shown utility
in quality control of commercially important botanicals like;
Ginseng, Andrographis paniculata, and echinacea. Although DNA
analysis is currently considered to be an innovative technology. It
has certain limitations due to which, its use has been limited to
academia. Another important issue is that DNA fingerprints will
remain the same irrespective of the plant parts used, while the
phytochemical content will vary with the physiology, plant parts
used, and environment.