UNIT-IV

Chapter 1
EVALUATION OF DRUGS

Syllabus content:
 WHO & ICH guidelines for the assessment of herbal drugs
 Stability testing of herbal drugs.
EVALUATION OF DRUGS
WHO guidelines for assessment of Herbal drugs:
 Assessment or evaluation of herbal drug means confirmation of its identity,
determination of its quality & purity and to detect the possible steps of adulteration to
ensure the genuine and true nature of the crude drugs.
 Evaluation of herbal drug is necessary because of three main reasons:
1) Biochemical variation in the drug
2) Deterioration due to improper processing and storage.
3) Adulteration and substitution.
 The WHO designed standard guidelines on quality control standardization of herbal
drugs. It was aimed to check the quality, safety and efficacy of herbal medicines
worldwide.
 Standardization of herbal materials is done in order to set a certain standard
monographic profile of any medicinal plants, which are having therapeutic value. This
will help to classify plants in Herbal Pharmacopoeia.
 Objectives of the WHO guidelines:
1. To guide principles for assessing the quality in relation to the safety of herbal
medicines with specific reference to contaminants and residues.
2. To provide model criteria to identify possible contaminants and residues.
3. To provide examples of methods and techniques.
4. To provide examples of practical technical procedures to control the quality of
finished herbal products.
WHO guidelines for assessment of Herbal drugs
WHO guidelines for assessment of Herbal drugs
STANDARDIZATION & QUALITY CONTROL EVALUATION OF HERBAL DRUGS
1. Authentication or identification:
 Identification of medicinal plants before processing
(Chemical/Pharmacological/Formulation) is one of the most important steps
for achieving a true material. Identification can be done by preparation of
herbarium sheets for identifying important taxonomical characters like family,
genus, species, morphological characters of the flowers, leaves, stem, presence
or absence of latex, gums, juice etc.
 Improper identification may lead to various problems such as mis-leading
plant identity, waste of time and money, submission of false report in Journals,
and may also lead to possible steps of adulteration.
 Govt. Institutes that identify plants: Botanical survey of India (BSI), Central
council for research in Ayurveda and Siddha (CCRAS), Central council for
Indian Medicine (CCIM), Pharmacopoeial Laboratory for Indian medicine
(PLIM), Central Institute of Medicinal and Aromatic plants (CIMAP), National
Botanical Research Institute (NBRI).
WHO guidelines for assessment of Herbal drugs

2. Organoleptic/Macroscopic/Morphology evaluation
 Here, identification is done by checking the external characters of the plant.
This is one of the easiest steps of evaluation since it gives a direct reflection of
how the plant material looks like.
 In this method of evaluation, various parameters can be used for ideintifying
the drugs such as Colour, odour, taste, shape, size, texture, fracture, extra
features etc.
 This is in fact the initial steps for identification of the plants in
Pharmacognostical aspects.
WHO guidelines for assessment of Herbal drugs
STANDARDIZATION & QUALITY CONTROL EVALUATION OF HERBAL DRUGS
3. Microscopic/Histology evaluation
 If two or more plant material are almost alike in external characters and
morphological identification seemed to be a problem for identification, it is to
be noted that identification of the closely related species can be done by
checking the microscopical characters.
 Qualitative microscopy: Transverse section & Longitudinal section
incorporating histo-chemical staining techniques (used to identify cellular
arrangements, identification of diagnostic characters, presence/absence of
lignification etc.), powder microscopy (Eg. types of trichomes, starch grains,
stomata, xylem vessels, oil glands, parenchymatous cells, tracheids etc.).
 Quantitative microscopy: Stomatal number, vein islet number, vein
termination number, starch grains number by Lycopodium spore method, size
of cells/trichomes/vessels etc.
WHO guidelines for assessment of Herbal drugs

4. Physical evaluation
 Physical evaluation deals with the physical parameters of the air dried plant
material. It directly reflects the purity of the sample.
 It is an important methods in plant identification since it report the amount in
percentage of the physical parameters of various characters such as :
a) Quantification of foreign matter
b) Quantification of moisture content (Loss on drying)
c) Determination of Ash value (Total ash, acid insoluble ash, water soluble
ash, sulphated ash)
d) Determination of extractive values (Alcohol and water extractive value)
e) Determination of volatile oil content
f) Determination of refractive index
g) Determination of optical rotation
WHO guidelines for assessment of Herbal drugs
5. Chemical evaluation
 This is one of the most important evaluation technique for proper
identification of plants since this technique is based on the determination of
chemical markers and the type of phytochemical classes present in the plant
material.
 This method also helps to identify the percentage of both major and minor
active phytoconstituents present in the plant extracts/material.
 Chemical evaluation can be categorized into two basic category:
1. Qualitative evaluation: This includes preliminary phytochemical
screening which detects what type of phytochemical classes are present in
the plant extracts. It also include thin layer chromatography techniques to
confirm the presence of a number of active phytoconstituents in the
extracts.
2. Quantitative evaluation: This is a higher level of evaluation of chemical
constituents in plants which includes spohisticated instrumentation
techniques such as HPTLC, HPLC, GC-MS, Chemical assays (like estimation
of total phenolic, total alkaloid, total tannin, total flavonoid, total steroid
content).
WHO guidelines for assessment of Herbal drugs

6. Biological evaluation
 Biological evaluation of medicinal plants is an important steps since it
evaluates all parameters which the plant material is directly or indirectly
having an impact associated with the human physiological system.
 There are three parameters to evaluate the biological process:
1. Determination of Microbial contamination : Total microbes viable
count, determination of pathogenic bacteria and determination of
aflatoxin content.
2. Pharmacological evaluation: Determination of bitterness value
(indications of bitter principles), determination of haemolytic index
(indications of saponins), foaming index (indication of saponins), swelling
index (indications of mucilage and gums).
3. Toxicological evaluations: Estimation of pesticides residues (both
chlorinated and phosphated pesticides), estimation of heavy metals (Pb,
Cd, As), estimation for the presence of radio active substances (actinides,
lanthanides) but only if recommended under strict Govt. regulations.
ICH Guidelines for Assessment of
Herbal Drugs
EVALUATION OF DRUGS
ICH guidelines for assessment of Herbal drugs:
 ICH stands for International Conference for Harmonization is an
international body which initiate and brings together regulatory authorities and
pharmaceutical industry to discuss scientific and technical aspects of
pharmaceutical product development and registration.
 Since its inception in 1990, ICH has gradually evolved, to respond to increasingly
global developments in the pharmaceutical sector and these ICH guidelines are
applied by a growing number of regulatory authorities.
ICH's mission is to achieve greater harmonization worldwide to ensure that safe,
effective and high quality medicines are developed, and registered and maintained
in the most resource efficient manner whilst meeting high standards.
 Herbal medicines acceptance globally is a big question, since it is composed of
numerous active phytoconsituents which gives problems when biological
evaluation is concerned like for example: Problem arise when evaluating the
pharmacokinetic parameters of a single extract bearing numerous compounds.
 Hence, ICH has design certain guidelines to overcome the problems and to
create impact on the regulatory assessment related to safety and efficacy of herbal
drugs for global acceptance.
EVALUATION OF DRUGS

4 basic ICH
WHO guidelines for assessment of Herbal drugs
1. Quality guidelines: Harmonization achievements on the quality area include
evaluating various parameters such as stability studies, defining relevant
thresholds for impurity testing and approach to pharmaceutical quality based
on Good Manufacturing Practices (GMP).
Parameters under ICH Quality guidelines:
Q1 Stability
Q2 Analytical Validation
Q3 Impurities
Q4 Pharmacopoeias
Q5 Quality of Biotechnological Products
Q6 Specifications
Q7 Good Manufacturing Practice
Q8 Pharmaceutical Development
Q9 Quality Risk Management
Q10 Pharmaceutical Quality System
Q11 Development and Manufacture of Drug Substances
Q12 Lifecycle Management
Q13 Continuous Manufacturing of Drug Substances and Drug Products
Q14 Analytical Procedure Development
WHO guidelines for assessment of Herbal drugs
2. Safety guidelines: There is a complete set of safety guidelines that ICH has
proposed to deal with health risk such as evaluation of carcinogenicity,
genotoxicity and reprotoxicity.
Parameters under ICH Quality guidelines:
S1 Carcinogenicity Studies
S2 Genotoxicity Studies
S3 Toxicokinetics and Pharmacokinetics
S4 Toxicity Testing
S5 Reproductive Toxicology
S6 Biotechnological Products
S7 Pharmacology Studies
S8 Immunotoxicology Studies
S9 Nonclinical Evaluation for Anticancer Pharmaceuticals
S10 Photosafety Evaluation
S11 Nonclinical Paediatric Safety
S12 Non-clinical Biodistribution Studies for Gene Therapy Products
WHO guidelines for assessment of Herbal drugs
3. Efficacy guidelines: These guidelines are concerned with the design, conduct, safety and
reporting of clinical trials data. It also covers novel type of medicines derived from
biotechnological processes and the use of pharmacogenetics/pharmacogenomics
techniques to produce better targetted medicines.
Parameters under ICH Effiacy guidelines:
E1 Clinical Safety for Drugs used in Long-Term Treatment
E2 Pharmacovigilance
E3 Clinical Study Reports
E4 Dose-Response Studies
E5 Ethnic Factors
E6 Good Clinical Practice
E7 Clinical Trials in Geriatric Population
E8 General Considerations for Clinical Trials
E9 Statistical Principles for Clinical Trials
E10 Choice of Control Group in Clinical Trials
E11 - E11A Clinical Trials in Pediatric Population
E12 Clinical Evaluation by Therapeutic Category
E14 Clinical Evaluation of QT
E15 Definitions in Pharmacogenetics / Pharmacogenomics
E16 Qualification of Genomic Biomarkers
E17 Multi-Regional Clinical Trials
E18 Genomic Sampling
E19 Safety Data Collection
E20 Adaptive Clinical Trials
WHO guidelines for assessment of Herbal drugs
4. Multidisciplinary guidelines: These are exceptional parameters which are apart from
quality, safety and efficacy guidelines. They include medical terminology for regulatory
authorities (MedDRA), common technical documents (CTD) and development of
Electronic Standards for the Transfer of Regulatory Information (ESTRI).
Parameters under ICH Multidisplinary guidelines:
M1 MedDRA Terminology
M2 Electronic Standards
M3 Nonclinical Safety Studies
M4 Common Technical Document
M5 Data Elements and Standards for Drug Dictionaries
M6 Gene Therapy
M7 Mutagenic impurities
M8 Electronic Common Technical Document (eCTD)
M9 Biopharmaceutics Classification System-based Biowaivers
M10 Bioanalytical Method Validation
M11 Clinical electronic Structured Harmonised Protocol (CeSHarP)
M12 Drug Interaction Studies
M13 Bioequivalence for Immediate-Release Solid Oral Dosage Forms
STABILITY TESTING OF HERBAL DRUGS
EVALUATION OF HERBAL DRUGS
Stability Testing of Herbal drugs:
 The concepts and principle of stability testing is to provide confirmation on how
the quality of an active pharmaceutical ingredient or medicinal products varies
with time under the influence of a variety of environmental factors such as
temperature, humidity, light, moisture and storage conditions.
 Since herbal drugs may contain single or more active constituents or
combination of herbs consisting of mixtures of phytoconstituents, hence stability
testing confining to a particular phytoconstituents is a problem.
 Therefore, evaluation of the available parameters based on chemical, physical,
microbiological, therapeutic and toxicological studies can serve as an important
tool in stability studies of herbals.
 In addition, worldwide stability testing of drugs happen to be tested in four
different climatic zones depending on the area where drug has been manufactured
1. Zone I: Temperate climate
2. Zone II: Subtropical, with possible high humidity
3. Zone III: Hot/Dry climate
4. Zone IV: Hot/Humid climate
EVALUATION OF HERBAL DRUGS
Factors affecting stability of herbal drugs:

Drug interactions
Storage conditions Content variations
Moisture content
Packaging
interaction
STABILITY
Mold growth Insect attack

Microorganisms Container material

Environmental factors
EVALUATION OF HERBAL DRUGS
Problems related to the Herbal Product Stability:
[1] Physical Instability: Natural medicines are usually prone to physical
instability due to the presence of impurities and reaction with the container.
Conditions like growth of microorganisms and insect feeding affect the
secondary metabolites and chemical composition of plants. Volatile active
components of natural medicine have the problem of volatility and decreasing
activity during storage for a long time.
[2] Environmental conditions: Condition such as fluctuation in rainfall, altitude,
temperature, light, soil, storage conditions as well as harvesting process, time and
method of collection and genetic variability can create substantial variability in the
product quality, stability and concentration of phytoconstituents.
[3] Chemical instability: Conditions that leads to chemical instability are mainly
related to chemical degradation (oxidation, hydrolysis, reduction) of herbals active
composition. Temperature and moisture are the two major important factors that
affect the quality and stability of herbal products. A chemical reaction may
increase by two or three fold for every 10ᵒC rise in temperature. On the other
hand, moisture is a good medium for increasing rate of decomposition and
hydrolysis. Presence of plant enzymes also accelerate the chemical degradation
process.
EVALUATION OF HERBAL DRUGS
Problems related to the Herbal Product Stability:
[4] Complexity, inconsistency and storage: Herbal formulations are complex
mixtures of different components of plants. Each plant components has variable
shelf-life, activity, concentration and consistency. This create problem during
storage condition, since it is not easy to determine the stability of each of the
phytoconstituents present in the herbal formulation, as each chemical compounds
present in the mixture has different stability conditions.
[5]Drug interaction, deterioration and decomposition : Every pahrmaceutical
formulation has to be packed in a suitable container for storage. There are a lot of
interaction happening between the low quality container with that of the active
ingredients of the formulation, this may lead to instability. In addition, interaction
may also takes place between the active medicaments with that of the excipients
added to the formulation which leads to instability.
EVALUATION OF HERBAL DRUGS
Role of markers in determining the stability of herbal drugs:
 Markers are chemically known compounds with a define structure, which may or
may not have therapeutic effect and are used to calculate the amount of herbal
medicinal ingredients in herbal products.
 Examples of some markers: Digitoxin from Digitalis, Ephedrine from Ephedra,
Piperine from Black pepper, Atropine from Belladona , Vit-C from Amla etc……
 Choice of the marker has to be justified and decision of the right analytical marker is
important for stability testing.
 Best Examples: Curcumin is the active chemical marker of Turmeric with a wide
range of therapeutic activity. To check the quality of any degradation or decomposition
of curcumin in a developed herbal product, curcumin composition is quantified using
HPTLC or HPLC. Any loss of curcumin content will directly reflect stability and efficacy.
 Another examples is instability of Ayurvedic formulation (Chyawanprash) which
may results in the degradation of Vitamin C, whereby Chyawanprash contain Amla fruit
pulp as the main ingredient.
 For an unknown medicinal plants, marker compounds may be isolated, purified and
structurally elucidated. The chemical structure of the isolated compound may be used
as a chemical marker for that plant. This activity not only helps to understand the
active principle of a plant, but also increases the analytical quality control process.
EVALUATION OF HERBAL DRUGS
Accelerated stability testing of herbal drugs:
 This is infact the most preferred method to test the stability of herbal products based on
storage condition.
Accelerated stability testing study predicts the SHELF LIFE of the product, by accelerating
the rate of decomposition, preferably by increasing the temperature of reaction conditions.
 Accelerated stability testing is sometimes referred to as stress stability testing and the
testing covers a time of up to 12 months simultaneously.
 Method of testing:
1. Take a known quantity of a drug formulation and place it in a container.
2. Measure all the initial physical parameters of the formulation (like pH, viscosity,
colour, odour, consistency etc.)
3. Subject the formulation to accelerated stability testing parameters (refer Table 1)
incorporating an Incubator with fixed temperature control. Sometimes in different
study testing the sample in varying temperature is also recommended (at 4ᵒC,
15ᵒC, 37ᵒC, 50ᵒC).
4. After the time period for stability testing is completed. Re-check the physical
parameters for any changes in stability.
5. If no change in initial and final results of the physical parameters, the formulation
is deemed considered to be stable under storage condition.
EVALUATION OF HERBAL DRUGS
Table 1: Storage conditions for stability testing of Drugs
Study Storage conditions Minimum time period coverd
by data at submission
Long Term Temperature: 25ᵒC ± 2ᵒC 12 months
Humidity: 5% RH
Intermediate Temperature: 30ᵒC ± 2ᵒC 6 months
Humidity: 5% RH
Accelerated Temperature: 40ᵒC ± 2ᵒC 6 months
Humidity: 5% RH