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Pharmacognosy Lab

Pharmacognosy is defined as an applied science which deals with the

biological, biochemical and economic features of natural drugs and their
constituents. It embraces knowledge of the history, distribution, cultivation,
collection, selection, preparation, commerce, identification, preservation and
use of drugs having their origin in the plants and animal kingdom.
Brief History of Pharmacognosy
In the early days, all information pertaining to drugs and their usage in
Western culture was designated materia medica (means medical matter).
The most famous commentary on drugs was written by Pedanios Dioscorides
entitled De Materia Medica libri cinque (Concerning Medical Matter) which
covered some 600 plant drugs plus a number of animal and mineral
products.
As the knowledge about drugs increased, specialized disciplines
became a necessity. In early 19 th century, materia medica began to undergo
division into pharmacology (the actions of drugs) and pharmacognosy (all
aspects of drugs with lesser emphasis on actions).
The word Pharmacognosyis from two Greek words pharmakon (drug)
and gnosis (knowledge).
This name was first thought to have been
introduced by Christianus Anotheus Seydler, a medical student in
Germany, who used the title Analectica Pharmacognistica in his dissertation
in 1815. However, recent historical research has found that the name was
earlier used by Johann Adam Schmidt in his Lehrbuch der Materia Medica
which was published in Vienna in 1811. It described the study of medicinal
plants and their properties.
Plants have become the major source of medicinal and pharmaceutical
substances. At present, people are going back to traditional medicine for the
treatment and prevention of diseases. Plant chemistry or phytochemistry is
an important element of pharmacognosy, different plant constituents have
their own distinct pharmacological actions.
Definition of terms
Materia Medica means medicinal materials. It is synonymous with
the substances and products derived from natural sources and employed by
the physicians in the old days.
Drugs are substances whether natural or synthetic, having therapeutic
or medicinal properties, and chiefly used as medicines or an ingredient in
medicine.

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Crude drugs are whole plants and animals and anatomical parts
thereof which have not undergone any process or treatment other than what
is essential to their proper packaging and storage. These are vegetable or
animal drugs which consist of natural substances that have undergone no
other process than collection and crying.
Drug constituents are chief constituents and their derivatives
obtained from drugs of biological origin or prepared synthetically or
semisynthetically.
Derivatives or Extractives are substances or mixture of substances
present in crude drugs which are separated by various means and are
employed in a more specific manner.
Natural products are any of the following:
Plant juices, exudates, secretions and extracts. e.g. aloe, acacia,
orange oil, etc.
Animal secretions and extracts. e.g. chymotrypsin, gelatin
Microbial extracts and products. e.g. xanthan gum, sutilains
Biologics are (a) products composed of antigenic matter or antibody
preparations capable of developing a state of immunity in the patient. e.g.
smallpox vaccine (b) diagnostic aids. e.g. tuberculin (c) related to human
blood. e.g. platelets, whole blood.
To evaluate a drug means to identify it and to determine its quality, purity
and pharmacological value. The evaluation of a drug involves a number of
methods which may be classified as follows:
1. Organoleptic evaluation makes use of the five senses.
The
appearance, feel (texture), smell (odor), taste and sound of materials
are noted. This evaluative tool is used for natural product and crude
drugs.

2. Microscopic evaluation makes use of the microscopic details of the

plant part such as leaves, stem and roots. Powdered materials are also
subjected to microscopic evaluation. Quality and size of particles are
important in the determination and differentiation of starches.
3. Physical evaluation makes use of physical constants for the
identification of the natural product. For instance, the identity and
purity of a fixed oil can be determined through its specific gravity as a

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different substance will give a different SG and impurities will tend to

change the SG of the oil. other physical parameters are melting point,
boiling point, refractive index, etc.
4. Instrumental evaluation is a more refined investigation of a natural
product. Infrared spectroscopy for example can reveal functional
groups that are present in the natural product, establishing its identity
and purity. Chromatography and spectroscopy are the most commonly
used methods for this evaluation.
5. Chemical evaluation uses test reagents for establishing the identity
of a natural product. Specific plant constituents will give specific color
reactions with test

6. reagents.
These reactions
may even differentiate the
same
class
of
plant
constituents.
7. Biological
evaluation
determines the effectiveness
of the plant constituent. Test
animals are employed to
establish its safety and
efficacy.
MICROSCOPIC ANALYSIS
microscopic characteristics and
histological cross-sections are
important.
Microscopes
are
used
to
observe hairs, vessels, esclereids,
stoma and acellular-like crystals
and grains of starch. This is used
when macroscopic evaluation is not
enough
to
differentiate
drug
products. (e.g. Histology for fresh
samples, particle evaluation for
powdered samples, etc.)

PHYSICAL
ANALYSIS

important in the establishment

of standards for identification
and quality of the drug product.
Physical parameters depend on
the drug sample used (crude or
natural product).

Humidity
Ash content/acid insoluble
ash content
Density
Refractive
index/optical
rotation
Foam index
Solubility
Melting point/boiling point
Other tests

CHEMICAL ANALYSIS

Phytochemical screening

Minerals
and
constituents

other

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Confirmation tests

Other special tests - Acid

value,
saponification
value

INSTRUMENTAL METHODS

Chromatographic
IR
NMR

BIOLOGICAL
Toxicological test
Pharmacological tests
- In vitro
- In vivo

EVALUATION OF PLANTS AND DRUGS

Plants are said to be indigenous if they grow in their native countries.
Plants that are only introduced and are grown in a foreign land is said to be
naturalized.
Plant materials may be collected from the wild or may be cultivated.
Cultivated plant materials have a greater advantage in that the growing
conditions are controlled. Optimum temperature, water supply and soil
nutrients ensure high quality of plant materials whereas the harsh conditions
in the wild may affect the quality and amount of plant constituents.
Here are the steps in the preparation of crude drugs for commercial
use:
Collection. The proper time of harvesting and collection is important
because the quality and quantity of plant constituents vary depending on the
time of day and season of the year.
Usually the most active plant
constituents are from mature plant parts and not the overly mature parts.
Harvesting.
The means of harvesting the plant part ensures is of
also of prime importance. Mass harvesting may be done by mechanical
devices but manual collection is always favored if the plant part is delicate.
Drying. The removal of moisture from the plant material will prevent
the growth of microbes such as molds and stops the action of enzymes that
may degrade the plant constituent of interest. The following are drying
techniques that may be employed.
Natural drying methods include exposing the plant part under
the sun or simply hanged to dry (air-dried). This poses
advantage in that the heat-labile materials are not destroyed
by the drying process. However, molds may grow because of
longer drying time.
Very plump plant parts are not
recommended for this type of drying as it will take more time.
Artificial drying methods include drying with the use of
heaters or the most common would be the oven. Care must
be observed so that the plant material is charred. Hence,
temperature must be controlled.
Garbling. This is the final step in the preparation of a crude drug. It
is the removal of extraneous matter such as dirt or other plant parts that
may have been collected with the plant part.

Packaging, storage and preservation. Appropriate containers for

packaging, storage conditions such as humidity and temperature will ensure
the preservation of the plant part.

ASH DETERMINATION
The ash content of a crude drug is generally taken to be the residue
remaining after incineration. The organic components of a crude drug are
combustible. Thus, the ash represents the inorganic salts naturally occurring
in the drug and adhering to it. These inorganic substances have little value
for purposes for evaluation. It may also contain the inorganic matter as
fraudulent adulterants to increase the weight of the crude drug. Different
materials would have different ash contents, thus a change in the ash
content of a crude drug may establish its identity and purity. The ash or
residue remaining after incineration of organic compound is a measure of the
amount of inorganic matter present as impurity. Addition of hot water is
employed for the removal of more polar components. Alcohol may also be
added and ignited. Usually what is left in the total ash are mainly carbonates,
phosphates, silicates and silica.
The total ash is the residue remaining after incineration. The acid
insoluble ash is the part of the total ash which is insoluble in diluted
hydrochloric acid.
MOISTURE CONTENT
Excessive water contained in drugs (e.g. aloes, gelatin, gums) are very uneconomical, and
possibly may also activate enzymes with a suitable temperature, causing the proliferation of
unwanted living organisms. Most vegetable drugs contain all essential food requirements for
molds, insects, and mites to cause deterioration rapidly once infected or infiltrated.
Loss on drying
Loss on drying is employed by the USP and BP/EP principally due to
small amounts of other volatile materials that contribute to the weight of a crude
drug or sample. These materials qualify as volatile in temperatures of 100-105C,
and the sample is dried until it reaches constant weight. Special considerations are
applied upon materials which require volatile substances such as balsams.
PHYTOCHEMICALS
Plants produce a vast number of substances of varying chemical structures.
These constituents may be present all throughout the plant tissue but may

also be stored in certain plant parts. There are two main groups of plant
constituents (phytochemicals)
Primary metabolites- these are relatively abundant in plants but
are of lower value. These include carbohydrates, proteins and
lipids.

Secondary metabolites- these are constituents that are present

in smaller quantities but possess higher pharmaceutical and
pharmacological value compared to primary metabolites. These
plant metabolites are also unique to particular species of plants.
Secondary metabolites include alkaloids, steroids, tannins,
flavonoids and others. These plant constituents may function for
plant reproduction (as pollinator attractants) or protection (as antifeedants/poison).
These plant constituents may be present in varying degrees in a single
plant which may hinder the efficient extraction of the plant constituent of
interest. Furthermore, they have exhibit different polarities, thus extracting
solvents vary depending on the plant constituent to be isolated.
PHYTOCHEMICAL SCREENING
Phytochemical screening is a tool used by phytochemists or natural
product chemists to evaluate the type of constituents present in plant
samples.
The methods used for screening should have the following
characteristics:
Simple
Rapid
Designed for minimum use of equipment
Reasonably selective for the class of compounds under study
Should provide additional information aside from the absence or
presence of the plant constituent under study.
Phytochemical screening is conducted on the crude ethanolic (for
nonpolar constituents) and aqueous (for polar constituents) extracts to
determine the presence of active principles in the plant material. This gives
the researcher a hint as to how the plant material is to be prepared, which
extracting solvent is to be used and what isolation technique is to be
performed.
Other problems that are likely to be encounterd are:

Difficulty to determine the presence of a particular class of compounds

due to a large number of diverse compounds occurring in the plant.
Uncertainty as to whether a substance is originally present in the plant
or simply a decomposition product due to the action of the light, heat,
acid, alkali or enzyme.
Varying composition of the plant part is not the same in all
circumstances.

KINDLY STUDY THE PHYTOCHEMICAL

SCREENING TESTS:
Phytochemical constituent and their specific
tests
Reagents used
Visual positive result (color, specific constituent
present)