Introduction of pharmacognosy & Phytochemistry

Introduction to
Pharmacognosy
UNIT-I
Mr. Vishal S. Bagul
Assistant Professor
Department of Pharmacognosy
H. R. Patel Institute of Pharmaceutical
Education and Research, Shirpur
(Maharashtra)
Mr. Vishal S Bagul, Assistant Professor., HRPIPER

Content
Definition, History, Scope and
Development of Pharmacognosy
Source of Drug
Organized Drug and unorganized Drug
Classification of Drug
Quality Control of Drugs of Natural Origin

Definition
• The word Pharmacognosy is derived from the
Greek "Pharmakon", meaning a drug or poison
and “gignosis" meaning to acquire knowledge
of and literally meaning “the entire knowledge
of drug”.
• The scientific study of the structural, physical,
chemical and sensory characters of crude drugs
of vegetable, animal and mineral origin.
• includes also their history, cultivation and
collection of the medicinal plants.

History
Hippocrates a Greek scientist, is known as the father of
medicine. He worked on human anatomy and Physiology,
particularly circulatory system and nervous system. He
prepared famous oath for physicians, which is still taken by
them.
Aristotle and Theophrastus well known philosopher and
scientist are known for their writing animal and plant kingdom
respectively.
• The practice of using herbs to cure disease is as old as
human civilization but a drastic change is observed in its
practice, as compared to that era.
• It is solely possible due to the contributions of many
known and unknown scientists.
• Plants were used medicinally in India, China, Egypt and
reporting or documenting the proof of herbs using
effectively.

Charak complied a group of ten herbs that
related to a certain disease. He made 50 such
groups which cover almost all the drugs
required by physicians for treatment. Then he
wrote the book Charak Samhita.
Sushruta arranged a large number of drugs into
distinct sets depending on their properties. He
was one who introduced the practice of surgery.
He arranged all his meticulous work into his
book Sushruta Samhita

Modern Pharmacognosy
• The development of modern Pharmacognosy began during the period of 1930-1960 by the
application of a broad spectrum of biological and socio-scientific subjects, including botany,
ethno botany, medical anthropology, marine biology, microbiology, herbal medicine, chemistry,
biotechnology, phytochemistry, pharmacology, pharmaceutics, clinical pharmacy and
pharmacy practice along with modern analytical techniques like paper and thin layer
chromatography (TLC), gas chromatography (GC), High performance liquid chromatography
(HPLC), Extreme and ultra-pressure liquid chromatography (XLC, UPLC), high pressure thin
layer chromatography (HPTLC), Mass spectroscopy, Liquid chromatography combined with
mass spectroscopy (LC/ MS), High Resolution Mass Spectroscopy (HRMS). Examples like
isolation of penicillin in 1928 by Alexander Fleming from microorganisms and later on
commercial production of the same in 1941 by Florey and Chain.

Name of Scientist and their
contributions
SCIENTIST INVENTION YEAR
Friedrich Adam
Isolation and Documentation of
Mesonic acid from Opium
1805
Karl L Reimann Isolation Nicotine from Tobacco 1828
Albert Niemann Isolation Cocaine from Coca Species 1860
Hardy Isolation Pilocarpine from Pilocarpus 1875
Nagayoshi Nagai Isolation Ephedrine from Ephedra 1885
Kiersten
Isolation Podophyllotoxin from
Podophyllum
1891

• Scope of Pharmacognosy

• Pharmacognosy is critical in development of different disciplines of science. A pharmacognosist should
possess a sound knowledge of the terms used to describe the vegetable and animal drugs as covered under
botany and zoology, respectively. The knowledge of plant taxonomy, plant breeding, and plant pathology
and plant genetics is helpful in the development of cultivation technology for medicinal and aromatic
plants. Plant - chemistry (phytochemistry) has undergone significant development in recent years as a
distinct discipline. It is concerned with the enormous variety of substances that are synthesized and
accumulated by plants and the structural elucidation of these substances. Extraction, isolation, purification
and characterization of phytochemicals from natural sources are important for advancement of medicine
system.
• Many of drugs today are being derived from herbs molecules.
• Many new molecules every year are being introduced through this field alone, so it is great source of lead
molecules.
• Marine sources possess about 5 Lakh species with lots of possibilities of developing medical purpose.
• Source of natural drug improves the importance of plants and animals.
• Beneficial against allopathic medicine because of less chances of toxicity issue to human being
• These are less cost effective formulations.

Development
of
Pharmacognosy
• Modern Pharmacognosy occurred during 1934 -
1960.
• This development was mainly due to some events
like
• Discovery of Penicillin in 1928 by Alexander Fleming
• Isolation of Reserpine 1952
• Anticancer properties of Vinca rosea
• Discovery of Dioscorea was utilized to prepare
semisynthetic
• Steroidal hormones.
• Pharmacognosy as an applied science.
• It played a crucial role in the development of
different disciplines of science.
• Some isolated components from plants were made
into different formulations and used as drugs

Applications of
Pharmacognosy
Plant bioactive Extraction and Isolation
• This type of approach is useful for the investigation of alkaloids,
antioxidants, bioflavonoids, terpenoids and other compounds as promising
therapeutics from an immense pool of plant biodiversity.
Pharmacognosy in drug development
• Pharmacognosy is used by pharmaceutical companies to screen,
characterize and produce new drugs for the treatment of human disease.
• naturally occurring drugs cannot be mass-produced, so they must be
studied in order to develop synthetic biosimilars.
Development of nano fertilizers and nano medicines:
• Nanotechnology in drug delivery is exemplified by nanocrystals, liposomes,
nanoparticle-protein conjugates, magnetic nanoparticles, nanogels and biodegradable
nanoparticles. Fertilizer particles can be coated with nano membranes that facilitate
slow and steady release of nutrients thereby reducing loss of nutrients and enhancing
its use efficiency of medicinal plants

• PLANT SOURCE
• Plant have always been a rich source of lead compounds (e.g. Alkaloids, Morphine, digitalis,
Quinine and nicotine).
• Almost all parts of the plants are used i.e. leaves, stem, bark, fruits and roots.
1. Leaves:
• The leaves of Digitalis Purpurea are the source of Digitoxin and Digoxin, which are cardiac
glycosides.
• Leaves of Eucalyptus give oil of Eucalyptus, which is important component of cold & cough
syrup.
• Tobacco leaves give nicotine.
• Atropa belladonna gives atropine.

Flowers: Poppy Papaver somniferum gives morphine (opoid)
Rose gives rose water used as tonic.
Fruits: Senna pod gives anthracine, which is a purgative.
fennel is a source of fenacone and it having carminative action.
Seeds: Seeds of Nux Vomica give strychnine, which is a CNS stimulant.
Castor seeds give castor oil.
Roots: Ipecacuanha root gives Emetine, used to induce vomiting as in
accidental poisoning. It also has amoebicidal properties.
Bark: Cinchona bark gives quinine and quinidine, which are antimalarial
drugs. Quinidine also has anti-arrythmic properties.
Stem: Ephedra stem contain ephedrine it use for appetite suppressant

ANIMAL SOURCE
• Pancreas is a source of Insulin, used in treatment of Diabetes.
• Sheep thyroid is a source of thyroxin, used in hypertension.
• Cod liver is used as a source of vitamin A and D.
• Blood of animals is used in preparation of vaccines.
• Stomach tissue contains pepsin and trypsin, which are digestive juices used in treatment of peptic
diseases in the past.
MARINE SOURCES
• The drug obtained from marine organisms which are being conventionally used like shark and cod-liver
oils, sodium alginate, agar-agar, chitin etc.
• Over 70% of the earth's surface is covered by oceans which contain 95% of the earth's biosphere.

TISSUE CULTURE
Is the growth of tissues or cells separate from the organism.
This is typically facilitated via use of a liquid, semi-solid, or
solid growth medium, such as broth or agar. Tissue culture
commonly refers to the culture of animal cells and tissues,
with the more specific term plant tissue culture being used
for plants.
The plant tissue culture technique is important because
isolation of bioactive compounds from the medium is very
easy, rare and endangered plant species are micro propagated
and cultivated in mass scale, production of immobilized plant
cell for future use and even biochemical conversion is easy.

• Organized drugs consist of the cellular organization in the
form of anatomical features. These are mostly the crude drugs
from plant sources.
• Microscopical and anatomical studies are preeminent for such
crude drugs. These can be used directly in medicine or can be
used by modifying or by extracting the active ingredient from
it.
Examples
• Bark: Cascara, Cassia, Cinchona
• Fruit: Anise, Capsicum, Caraway
• Seeds: Black Mustard, Cardamom, Nux-vomica
• Roots and Rhizomes: Aconite, Garlic, Ginger, Gingsing
• The unorganized drugs do not have the morphological or
anatomical organization as such. These are the products
which come directly in the market but their ultimate source
remains the plants, animals or minerals.
• These products may be solid, semisolid or liquid and the
physical, chemical and analytical standards may be applied
for testing their quality and purity.
Examples
• Dried Extract: Agar, Black catachu, Pale catachu, Pectin
• Waxes: Beeswax, Spermaceti
• Gums: Acacia, Gaur gum
• Volatile oil: Coriander, peppermint, Sandalwood
ORGANIZED DRUG
UNORGANIZED DRUG

CLASSIFICATION
OFDRUGS
Alphabetical classification
Morphological classification
Pharmacological classification
Chemical classification
Taxonomical classification
Chemotaxonomic classification
Sero-taxonomical classification

Alphabetical classification
• Crude drugs are arranged in alphabetical order of their Latin and English names
(common names) or sometimes local language names (vernacular names).
• Some of the pharmacopoeias, dictionaries and reference books which classify crude drugs
according to this system are as follows:
1. Indian Pharmacopoeia
2. British Pharmacopoeia
3. British Herbal Pharmacopoeia
4. United States Pharmacopoeia and National Formulary
5. British Pharmaceutical Codex
• In European Pharmacopoeia these are arranged according to their names in Latin where in
United States Pharmacopoeia (U.S.P.) and British Pharmaceutical Codex (B.P.C.), these
are arranged in English.
Example: Acacia, Bees wax, Castor oil, Datura …….etc Mr. Vishal S Bagul, Assistant Professor., HRPIPER

Advantages
• It is very simple to use and
operate.
• It is very easy to find out
the specific data and go to a
specific entry.
• There is no repetition of
single entry.
Disadvantages
• It does not give any
idea regarding
chemical constituents,
uses and other
information regarding
plants.

Morphological classification
• Plants are made up of different parts. Different plants can be grouped according to
the part which is being utilized . This system is truly useful, as it gives guidance
regarding the harvesting of plants.
E.g. Leaves: Vasaka, Senna, Digitalis
Bark : Cinchona, Arjuna, Cinnamon
Roots : Rauwolfia, Ashwagandha
Fruits : Coriander, Fennel
Whole plant: Tulsi, Vinca
Latex : Opium

Pharmacological classification:
In this system of classification, the plants are arranged accordance with their pharmacological or
biological activity. Different plants give certain medicinal effects
E.g. Anti-diabetic: Fenugreek, Neem
Anti-cancer: Taxus, Vinca
Anti-malarial: Cinchona, Artemisia
Advantages:
It prepare a list of substitutes which can be utilized for treating same condition.
Disadvantages :
1.Some plant possess more than one activity; in such cases the plant name is repeated.
2 .Pharmacological classification is unable to give data regarding parts of plants being used
especially in case of unorganized drug, it create problems.

Chemical classification:
• Depending upon the active constituents, the crude drugs are classified. The plants
contain various constituents in them like alkaloids, glycosides, tannins,
carbohydrates, saponins, etc. Irrespective of the morphological or taxonomical
characters, the drugs with similar chemical constituents are grouped into the same
group.
• Therapeutics effects observed in plants, only depends on the chemical constituents. The
presence of one of the chemical constituents can be correlated to certain specific
biological effects.

• Advantages:
a. It arranges the plant according to their chemicals,
which after flexibility to study further subtypes.
b. For studying the plants, this is the best method.
c. Due to similar chemical classes many times similar
extraction and isolation procedure can be employed.
E.g. Stas and Otto
• Disadvantages
It does not resemble to any pharmacological benefits of
plants.

Taxonomical classification
This is also referred to as Biological classification. It is an old system
initiated by Linax. In this method plants are mentioned with various
ranked levels.
The levels contain plants with some similar morphological
properties. This approach of classification allows us to understand
various correlations among the various plant species.
Kingdom: Plantae-It contains all sort of plants, green algae, ferns etc.
Division: Angiospermae- Flowering plants
Class: Dicotyledons- Seeds can be divided in two equal parts.
Order: Rosales- Flower producing order
Family: Leguminosae- Nitrogen fixing bacteria in roots
Genus : Trigonella- Leaves appear in groups of three

1. We can study the different drugs with
same family at a time due to its resemble
characteristics of that particular family.
2. Detail botanical information and
evolutionary development of plant can be
studied.
Advantages
Disadvantages
1. It fails to recognise organised and
unorganised drugs………………..………….
2. Many drugs are not entire plants and
represent part of plant that have been
processed systematically that are not
classified by this system
3. It fails to give the data of chemical nature
of active constituent and therapeutic
significance of crude drug.

Chemotaxonomic classification:
• This system of classification on the chemical similarity of a taxon, i.e. it is based on
the existence of relationship between constituents in various plants.
• There are certain types of chemical constituents that characterize certain classes of
plants. This gives birth to entirely a new concept of chemotaxonomy that utilizes
chemical facts/characters for understanding the taxonomical status, relationships and
the evolution of the plants.
• For example, tropane alkaloids generally occur among the members of Solanaceae,
thereby, serving as a chemotaxonomic marker. Similarly, other secondary plant
metabolites can serve as the basis of classification of crude drugs.

Advantages
• It gives more scope for understanding the relationship between
chemical constituents,
• their biosynthesis and their possible action.
• The characters most often studied are secondary metabolites
of pharmaceutical
• significance such as alkaloids, glycosides, flavonoids, DNA
hybridization, amino acid
• sequencing in proteins etc.
• It provides degree of hybridization and breeding analysis
Disadvantages:
• This system is fails to identify the organized and unorganized
crude drugs in their morphological studies.
• This system fails to understand the therapeutic nature of the
crude drugs.

Serotaxonomical
classification
The study of antigen-antibody reaction is called serology.
The application of serology in solving taxonomic
problems is called Serotaxonomy.
The technique is based on the highly specific relationship
between antigens and antibodies produced in response to
them.
It expresses the similarities and the dissimilarities among
different taxa, and these data are helpful in taxonomy.
It determines the degree of similarity between species,
genera, family, etc., by comparing the reaction with
antigens from various plant taxa with antibodies present
against a given taxon.

Gum and Mucilage
Gum
• is a sap or other resinous material associated with certain
species of the plant kingdom. This material is often
polysaccharide-based and is most frequently associated with
woody plants, particularly under the bark or as a seed coating.
The polysaccharide material is typically of high molecular
weight and most often highly hydrophilic or hydro colloidal.
Mucilage
• a polysaccharide substance extracted as a viscous or gelatinous
solution from plant roots, seeds, etc., and used in medicines
and adhesives.

Mr.
Vishal
S
Bagul,
Assistant
Professor.,
HRPIPER
Dried Latex
• is a stable dispersion
(emulsion) of polymer micro
particles in an aqueous
medium.
Resins
• Resin is a solid or highly
viscous substance of plant or
synthetic origin that is typically
convertible into polymers.
Oleo resin-
• A natural or artificial mixture
of essential oils and a resin,
e.g. balsam.
Oleo gum resin-
• a solid plant exudation (as
asafetida) consisting of a
mixture of volatile oil, gum,
and resin

Quality control of Drugs
of Natural Origin
• Adulteration of drugs of natural origin.
• Evaluation by organoleptic, microscopic,
physical, chemical and biological
methods and properties.
• Quantitative microscopy of crude drugs
including lycopodium spore method, leaf
constants,
• Camera lucida and diagrams of
microscopic objects to scale with camera
lucida

Quality
Control
As per Pharmacopeial definition, the quality control of
herbal drugs are based on three terms likely Identity, Purity
and Assay.
Identity 2. Purity 3. Assay
Quality of herbal drugs are defined as the status of
a drug that is determined by its evaluation in terms
of identity, purity, content and other physical,
chemical and biological properties and
manufacturing processes for formulated drugs.

Adulteration
• It is a practice of substituting original crude drug partially or wholly with similar
looking substances.
Reason For Adulteration
• To get more profit.
• Due to shortage of drugs.
• In case of costlier drug.
• For banned drugs

1. Substitution with artificially manufactured substances
Such a type of adulteration is found in case of costlier
drugs. Some plants or parts of plants rarely available
because of the season or climatic conditions. Some artificial
substances with similar looks are available at cheaper rate.
Example: Artificial invert sugar for Honey
2. Uncertain identity of the drug
Sometimes due to the confusion of the authenticity of the
plant, drug identity becomes uncertain. Like Chichona bark
it has different species like Cinchona calisaya, C.
officinalis, C. ledgeriana etc. are considered.

3 Seasonal availability of drugs: Some drugs are available in a specific season so other
drugs can be introduced in their absence which has same action. For example, Trianthema
portulacastrum can be used in seasonal absence of Boerhavia diffusa
4 Harmful adulterants: This is happened with unorganized and liquid drugs. Wastes
materials are collected and mixed with original drugs. Eg. Pieces of amber colour glass with
colophony.
5. Substitution with totally different drugs: This practice is generally done in case of oil.
Like cotton seed oil in place of olive oil. Sometimes barks also substitute original bark.
6. Substitution of species in the same family: This practice is done in case of dried leaves,
roots or stems. Example: Dog senna in place of Indian seena, but both have same family
(Leguminosae)

7. Substitution with exhausted drugs- Volatile oil containing drugs
are generally adulterated by method same drug are being used as an
adulterants but they do not contain medicinally active constituents as
they are already extracted out. Some time additives are added to impart
colour as well as taste. Original tea powder and filtered tea powder
after use look similar but chemical constituent from the filtered tea
powder have been extracted and missing Eg – Fennel, Coriander etc

Quality Control
a system of maintaining standards in manufactured
products by testing a sample of the output against the
specification.
Evaluation
Confirmation of its identity and determination of its
Quality and Purity and Detection of nature of adulterant by
various parameters like
Morphological, Microscopical, Physical, Chemical and
Biological observations

MORPHOLOGICAL / ORGANOLEPTIC
• Organoleptic evaluation of crude drugs refers to the evaluation of a drug by colour, odour, taste, size and
shape, occasionally the sound or snap of fracture and special fetures including touch, texture, etc..
• It is a technique of qualitative evaluation based on study of morphological and sensory profiles of whole
drugs.
Advantage: It provides simplest and quickest means to establish the identity & purity and thereby ensure
quality of particular sample
Disadvantage: Judgement may vary from person to person and time to time.
Example:
• Fractures of barks like cinchona, cascara.
• Sweet taste of liquorice

BOTANICAL
MACROSCOPIC
• Examine the untreated sample. If necessary, a magnifying lens (6x to 10x) may be used. Wetting with
water or reagents, as required, may be necessary to observed the characteristics of a cut surface.
• The texture is best examined by taking a small quantity of material and rubbing it between the thumb and
forefinger, it is usually described as ‘smooth’, ‘rough’
a) Shape and size
b) Colour and external marking
c) Fracture and internal colour
d) Odour and taste

MICROSCOPICAL
• Leaf constants or diagnostic characters of leaf-
• Stomatal numbers-
• Is average numbers of stomata per.sq.mm of epidermis of leaf.
• Stomatal Index-
• Is the percentage which number of stomata form to the total
number of epidermal cells, each stoma being counted as one
cell.
Epidermal cell

Vein –Islet number-
• It is defined as number of vein-islet present per square
mm area of leaf surface
Vein termination number-
• It is number of vein terminating at particular end per
square mm area of leaf surface

Calcium Oxalate Crystal:
• Calcium oxalate crystals are present in almost all parts
of the plant. The Calcium oxalate crystals are either the
monohydrate whewellite form or the dihydrate
weddellite form. They are identified by X-ray
diffraction; Raman microprobe analysis and infrared
spectroscopy are the most accurate. Under microscope
they are visible as colorless when treated with chloral
hydrate solution.
Functions:
• They give protection to the plant against environmental stress.
• They help in identification of crude drugs.
• They help in detection of adulterants.
• They help in identification of plants in same family.

Trichomes
• Trichomes is the term used to refer to tiny outgrowths from the plant epidermis. Although the term
"trichomes" generally refers to outgrowths ranging from small hairs to larger outgrowths like thorns, it is
typically used to refer to the tiny hairs that can be seen emerging from the surfaces of leaves and other epidermal
surfaces of plants. Trichomes are classified in main three types on the basis of structure and number cells present
1) Covering or non-globular or clothing trichomes- eg., Nux-vomica, Tea, Cannabis.
2) Glandular trichomes- eg., Vasaka, Piper, Digitalis.
3) Hydathodes or special type of trichomes- eg., Piper betal, London pride.
Functions of Trichomes:
1. Plant defense against insects
2. Generally a dense covering of woolly trichomes controls the rate of transpiration.
3. They also reduce the heating effect of sunlight.
4. They aid in the protection of plant body from outer injurious agencies.
5. Chemicals produced in the glandular tip can deter feeding or the trichome can physically
prevent the insect from reaching and feeding on the leaf.

PHYSICAL
Qualitative
1. Refractive Index-
• When a ray of light passes from one medium to another of
different density, it is bent from original path. Thus, the ratio
of the velocity of light in vacuum to its velocity in the
substance is termed as refractive index of the second
medium. Depending upon purity it is constant for a liquid.
Eg- Castor oil 1.4758-1.4798

2. Viscosity:
• Viscosity is a measure of the resistance of a fluid which is being deformed by
either shear or tensile stress. The most common method of determining
kinematic viscosity in the lab utilizes the capillary tube viscometer.
Examples: Clove oil: 0° to – 1.5°, Cinnamon oil: 0° to – 2°.
3. Foreign Organic Matters:
• Anything extra present in the drug which is not complying with the authentic
drug may be considered as a foreign matter. The foreign matter can be present in
the drug due to improper harvesting. The source of foreign organic matter can be
animal excreta, insect or mould and is determined by sedimentation or floatation
method.
• It determines presence of organic as well as inorganic foreign matter.
• It gives an idea weather drug is pure or adulterated
Examples: Curcumin – not more than 2.0%; Neem – not more than 1.5%

4. Solubility-
• The presence of adulterants can be determined by solubility studies.
• Examples: Castor oil is soluble in 3 volumes of 90 percent
alcohol, while the adulterated form may shows good solubility in
alcohol.
5. Extractive Values:
• All the chemical constituents are soluble either in polar, semi polar or
organic solvents. Total soluble constituents of the drug in any particular
solvent or mixture of solvents may be called its extractive value or
percent extractive.
Significances of Extractive Values:
• This method is important when the constituents of drugs can’t be
readily estimated by any other means.
• It indicates the nature of chemical constituents present in drugs.
• It helps in identification of adulterants.

6. Ash value: Ash value is useful
in determining authenticity and
purity of sample and also these
values are important qualitative
standards.
• Acid insoluble ash value
indicates siliceous impurities.
• Water soluble ash value gives
an estimation of inorganic
contents

7. Moisture content
Moisture content refers to the number of
water molecules that become incorporated
into a Crude and food product.
8. Swelling index
The swelling index is the volume in ml taken
up by the swelling of 1g of plant material
under specified condition. Its determination
is based on the addition of water or a
swelling agent as specified in the test
procedure for each individual plant material.

• Plant materials containing saponins cause
persistent foam when the decoction is
shaken. Foaming ability of plant material
and their extracts is measured by foaming
index
9. Foaming Index:

CHEMICAL
• It is qualitative as well as quantitative type of evaluation.
• Chemical tests and chemical assays are prime important.
• Isolation, purification, and identification of active constituents .
• Preliminary phytochemical screening is qualitative evaluation.
• Qualitative chemical tests are useful in detection of adulteration.
• eg., Copper acetate is used for detection of balsams and waxes.
Qualitative

• Test for alkaloid
Dragendoff’s test
Mayers test
Hagars test
Wagnar test
• Test for carbohydrates
Molisch test
Barfoed’s test

QUANTITATIVE
• Colorimetric Method: It is a method of determining the concentration of a chemical
element or chemical compound in a solution with the aid of a color reagent. It is
applicable to both organic and inorganic compounds and may be used with or without an
enzymatic stage. The method is widely used in medical laboratories and for industrial
purposes, e.g., the analysis of water samples in connection with industrial water
treatment.
• UV-Visible Spectroscopy

• Volumetric Method: It is a quantitative analysis
of liquids or solutions by comparing the volumes
that react with known volumes of standard
reagents, usually by titration. A reagent is prepared
as a standard solution, acts as titrator. A known
concentration and volume of titrant reacts with a
solution of analyte or titrand to determine
concentration.

• Photometric Method: It is the set of methods of
quantitative chemical analysis based on the
relationship between the concentration of a
substance in a solution or gas and the absorption
of radiation. In this method the intensities of the
monochromatic components of transmitted
radiation are scanned

• Gravimetric Method: It is the quantitative determination of a
substance by the precipitation method of gravimetric analysis involving
isolation of an ion in solution by precipitation reaction, filtering,
washing the precipitate, conversion of precipitate to a product of known
composition, and finally weighing the precipitate and determining its
mass by difference. There are four fundamental types of gravimetric
analysis: physical gravimetry, thermogravimetry, precipitative
gravimetric analysis, and electrodeposition.

BIOLOGICAL
• Measurement of biological effect and
Pharmacological activity of new or chemically
undefined substance.
• Determination of the side effect profile, including
the degree of the drug toxicity.
• Measurement of the concentration of know
substance
Cardiac glycoside are evaluated by this method
using frogs, cats, pigeons

Microbiological assay
• The method of measuring compounds such as
vitamins and amino acids using microorganism.
• As per IP, two methods are used for the assay
• Cylinder plate method or Cup Plate methods
Cylinder plate method: based upon the diffusion of
the antibiotic throughout solid culture media.
Cup Plate methods: antibiotic containing cylinder is
diffused into the agar layer containing the
microorganisms

Analytical Methods
• In this method various chromatographic
technique as well as spectroscopic
methods are used for the sample.
Thin Layer chromatography (TLC):
used for separation of Mixture with the
principal of adsorption.

High Performance Thin Layer
Chromatography (HPTLC)
• This system is a versatile modern analytical
technique with reference to its excellent
automation, optimization, multidimensional
application.

High performance Liquid chromatography
(HPLC)
• It is highly improved column
chromatography. By this methods
solvent allowed passing though the
stationary column chromatography.

Gas Chromatography (GC)
• It is used for separation and
analysis of vaporized compound
which are non decomposable.
• Nitrogen gas are mostly used
for this chromatography

UV-Visible Spectroscopy
• This method refer to absorption
spectroscopy in the visible spectral region.
• UV range id 800-200nm
• Morphine is determined at 286nm

Infra-red Spectroscopy (IR)
• It is used for identification of
functional group and structure
elucidation, identification of
substance, detection of impurities.
• (14000-4000 cm-1)

Lycopodium spore method
• This method is used to identify the crude drugs when the chemical and physical methods
are inapplicable.
• This method is also useful to detect the adulteration present in the crude drugs
containing starch grains.
• Lycopodium spore method was performed on ingredients of Shatavar churna, an
ayurvedic formulation used as immunomodulator
• Examples: Adulterated drug containing starch can be determined by counting the
number of starch grains per mg and calculating the amount from the known number of
starch grains per mg of the pure starch. The percentage purity of an authentic powdered
ginger is calculated using the following equation:

Significance:
• Determination of foreign organic matter.
• Determination of percentage purity of drugs.
• Detection of adulterant

Leaf constants
• Important quantitative Microscopic evaluations which can be used to identify
and distinguish between some closely related species.
• Stomata
• Stomatal numbers
• Stomatal Index
• Vein –Islet number
• Vein termination number-
• Calcium Oxalate Crystal
• Trichomes
• Palisade ratio

CAMERA LUCIDA
• It is an optical device or instrument in which rays of light are reflected by a prism to
produce an image on a sheet of paper, from which a drawing is made. It works on simple
optical principle reflecting beam of light through a prism and a plane mirror.

• Camera Lucida, when attached with a compound microscope, helps
drawing microscope images of objects on paper.
• It works on simple optical principle reflecting beam of light through a
prism and a plane mirror. The microscopic image of the object is
reflected by the prism on to the plane mirror and there from the
image is reflected on to the plane paper.
• The observer moves the pencil on the lines of the image and draws a
correct and faithful figure of the object on the paper. There are three
main parts of a camera lucida the attachment ring, the prism, and the
mirror.
• The attachment ring attaches the camera lucida with the body tube of
the microscope. The prism rests just above the eyepiece when the
instrument is attached with the microscope.

Introduction of pharmacognosy & Phytochemistry

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