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Pharmacognosy & Phytochemistry I
BP405T-1
Ms. Sapna kumari
Assistant Professor
School of Pharmacy
Introduction to Pharmacognosy
Classification of drugs
Quality Control of Drugs of Natural origin
CHAPTER-1
Definition, History, scope and
development of Pharmacognosy
A
Definition
• Study of crude drugs obtained from plant, animals and minerals
and study of their active constituents.
• Derived from Latin term
• Pharmakon- A drug
• Gignosco- To Acquire knowledge of
• Pharmacognosy word first used as Pharmacognosis.
• Term pharmacognosy first used by John .Adam .Schmidt, A
Austrian Physicist and he used the term Pharmacognosist in 1811
in his work Lehrbuch der Materia Medica“.
• After J. A. Schmidt, C.A Seydler used the term Pharmacognosis in
his work Anlactica Pharmacognostica in 1815.
WHAT IS PHARMACOGNOSY?
• Pharmacognosy is the study of crude drugs from natural sources
including information regarding
History,
Distribution,
Cultivation & collection
Sensory, physical and physicochemical characterization
Structural features (Phytochemical Analysis)
Medicinal uses,
Processing of crude drug for commercial market
Storage
Preservation of drug.
AD
(ANNO
DOMINI)
In the year of
lord
BC
(BEFORE CHRIST)
0
BIRTH
OF
JESUS
CHRIST
 Concept of AD and BC was given by POP DIONYSIUS, he was a Christian.
Before the
year
of Christ
History
In India medicinal properties of
plants described in
Rigveda and Ayurveda (3500-
1500 B.C.)
Charaka Samhita – plants
Sushruta Samhita-surgery
Papyrus Eber (1500 B.C.)- it is an
oldest written document of egypt
it contains 876 formulations and
700 crude drugs.
In china medicinal plants had been
used since 5000 B.C. Egyptians (Ebers papyrus, 1550 BC)
History
Hippocrates –Father of medicine (460-360 BC)
• He was a Greek scientist and has described human anatomy
and physiology.
• He has also describe collection, identification, uses, property,
of around 300 drugs.
• Aristotle- Father of biology(384-322 BC)
• Student of Pluto & known for his writing on animal kingdom.
• Theophrastus - Father of botany (384-322 BC)
• Student of Aristotle wrote about plant kingdom.
• Galen- Described method of preparations of plant and animal
drugs known as “Galenicals”
History
• Dioscorides
• “De Materia Medica” (600 medicinal plants)
• Described some medicinal plants like Belladona, Ergot, Opium,
Colchicum etc.
History
• Swede linnaeus –Classified plants and introduced binomial system for
naming plants.
• It is a system used to name species
• Eg- genus-homo, species-sepiens
• To ensure that scientific name of an organism is same all over the
world.
• Classification further developed by Bentham & Hooker-
Monocotyledon, dicotyledon & gymnosperm.
• Eicher & Engler.
• Mendel – Study on hybrids.
• Paracelus – Developed mineral salts.
• Le’mary- Importance of extraction method and alcohol as extractant.
• Willium withering – Published some of medicinal properties of fox
glove leaves.
• Stass & otto- Developed extraction process for alkaloids.
Development
• 1934-1960
• Development of modern pharmacognosy with integration of chemistry,
biotechnology, analysis, cology etc.
• Around 75 substance from plant were isolated and structures were determined
along with pharmacological studies.
1928 Penicillin Antibiotic
1952 Reserpine Hypotensive and
tranquilizer( drug
reduce reduce tension)
1955 Vincrisitin and vinblastin Anticancer
Blood and lymphcancer
1960 Diosgenin Preparation of sterodal
hormones
Development
• 1960-1970
• Antibiotics like
• Streptomycin,
• Chloramphenicol,
• Tetracycline ,
• Gresiofulvin etc isolated so called antibiotic age.
• Phytoconstituents from
• Digitalis
• Senna
• Aloe
• Belladona
• Ergot were isolated and their clinical used were established
Development
• SAR
• Semi synthetic preparations
• E.g. Ergometrin Methyl Ergometrin Better oxitocic activity
• Study of biosynthetic pathways
• Shikimmic acid & other biosynthetic path way
Constituents Acivity Responsible group
Reserpine Hypotensive Trimethoxy benzoic
acid
Tubocurarine Muscle relaxant Quaternary
ammonium group
Development
• Natural molecule as model for synthesis of new drug
• E.g. During world war –II Quinine as an antimalarial drug was short in supply so
similar type of compound chloroquine was isolated
• Cocaine to lignocaine, xylocaine prepared as local analgesic
• Silicin to salicylic acid (Aspirin) prepared which is used as Analgesic, Anti
thrombolytic and Anti inflammatory agent.
Development
• 1970-1990
Phytoconstituents Plant Name Activity
Podophllotoxin
(Etoposide, Teniposide)
Podophyllum Lung and testicular
cancer
Silymarin Milk thistle Liver disease
Taxol Taxus Ovarian, Breast and lung
cancer
Guggulosterol Guggul Arthritis,
Hypercholesteremia
Ginkolide Ginko Increase blood circulation in
brain
Ginsenoside Ginseng Tonic
Development
• 1990 onwards
 Herbal drug demand increase due to less toxicity, side effect and cost.
 To control quality WHO published guideline and prepared monograph.
 Gov. Of India published Ayurvedic pharmacopoeia part I &II.
 Indian drug manufacturers published 3 volume of Indian herbal
pharmacopoeia.
 USP 2010- 18 plants
 IP 2009 - 27 plants
 TKDL – Traditional knowledge digital library
 Prepared by india from 2 feb 2009 to protect traditional knowledge.
 7 crore TKDL project took 9 year to enlist 2 lakh formulations from
Ayurveda Siddha and unani.
 Available in english, German, Japaniese, french and spanish.
Scope
• Identification of proper drug by morphology and microscopy.
• Quality of herbs known by evaluation.
• Good quality of drug obtained by proper cultivation and
harvesting, drying & storing at proper time.
• Cultivation of drug which are in demand and earn by selling it.
• Active constituents are isolated from plants and can be marked
to earn.
• Pharmacognosy is an important link between Pharmacology and
Medicinal chemistry.
• Crude drugs also provides the intermediate precursor for synthesis
of new drug.
• Better active constituent’s semisynthetic derivatives can be
prepared.
• Eg. Ergotamine to Methyl Ergotamine
• Synthetic molecule based on plant active constituent provide
better activity.
Eg. Quinine –Chloroquine (Antimalarial activity) Vasicine –
Bromohexine (Bronchodilator)
Scope
• we can obtained new medicine for new diseases like AIDS, Cancer
etc.
• Plant tissue culture and genetic engineering gives more advance
natural products.
• Natural products as Pharmaceutical aid.
Accacia & tragacanth – Binding, emulsifying & suspending agent.
Carbohydrates- Sweetening agent
Volatile oil – Flavoring agent
Fixed oil- Ointment base, emolient
Starch- Disintigrating agent
Piperine- Bioavaibility enhancer.
Scope
Sources of drugs
B
Exploration of medicinal plant in
ancient time
• Trial and error- try and discard until its serves
the purpose
• While searching for food- herbs and species
are identified.
• Signature of nature- similarity between
human organ and plant part
• Zoo pharmacognosy-animal self medication
power
• Accidental discovery- cinchona,penicillin
• Drugs are substances that are used or intended to be used in the
diagnosis, prevention, treatment or cure of diseases. In early times, these
substances were derived from natural sources, of which plants took up the
major share. With the introduction of technology, most drugs today are
manufactured synthetically in the laboratory. The major sources of drugs
can be grouped into the following
1. Plant source
2. Animal source
3. Microbial source
4. Mineral Source
5. Marine source
6. Tissue culture
Sources of drug
Plant Source
Plant source is the oldest source of drugs. Most of the drugs in
ancient times were derived from plants. Almost all parts of the
plants are used i.e. leaves, stem, bark, fruits and roots.
Leaves:
a. The leaves of Digitalis Purpurea are the source of Digitoxin and
Digoxin, which are cardiac glycosides.
b. Leaves of Eucalyptus give oil of Eucalyptus, which is important
component
Sources of drug
Plant Source
c. Tobacco leaves give nicotine.
d. Atropa belladonna gives atropine.
Flowers:
Poppy papaver somniferum gives morphine (opoid)
Vinca rosea gives vincristine and vinblastine
Rose gives rose water used as tonic.
Photo of Papaver somniferum by Evelyn Simak
Fruits:
Senna pod gives anthracine, which is a purgative (used in constipation)
Calabar beans give physostigmine, which is cholinomimetic agent.
of cough syrup.
Sources of drug
• Seeds:
• Seeds of Nux Vomica give strychnine, which is a CNS stimulant.
• Castor oil seeds give castor oil.
• Calabar beans give Physostigmine, which is a cholinomimetic drug.
• Roots:
• Ipecacuanha root gives Emetine, used to induce vomiting as in accidental
poisoning. It also has amoebicidal properties.
• Rauwolfia serpentina gives reserpine, a hypotensive agent.
• Reserpine was used for hypertension treatment.
• syrup.
• Bark:
• Cinchona bark gives quinine and quinidine, which are antimalarial drugs. Quinidine
also has antiarrythmic properties.
• Atropa belladonna gives atropine, which is anticholinergic.
• Hyoscyamus Niger gives Hyosine, which is also anticholinergic.
Sources of drug
Animal source
• Drugs obtained from animals sources are whole animals, glandular products
(thyroid organ), liver extract, polypeptide venoms, non-peptide toxins, etc. Fish
liver oil, musk, beeswax, hormones, enzymes, and antitoxins sera are the
products obtained from animal sources (kokate). A large number of other natural
products from animal sources are used as pharmaceutic excipient and others are
used as important drugs or as nutritional supplements. The examples are as
given below (Annamalaiuniversity):
• Pancreas is a source of Insulin, used in treatment of Diabetes.
• Urine of pregnant women gives human chorionic gonadotropin (hCG) used
for the treatment of infertility.
• Sheep thyroid is a source of thyroxin, used in hypertension.
Sources of drug
•Cod liver is used as a source of vitamin A and D.
•Anterior pituitary is a source of pituitary gonadotropins,
used in treatment of infertility.
•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. Nowadays better drugs have replaced them.
•cough syrup.
Sources of drug
Microbial Source
Many life-saving drugs are obtained from microbes,
Penicillium notatum is a fungus which gives penicillin.
Actinobacteria give Streptomycin.
Aminoglycosides such as gentamicin and tobramycin are
obtained from streptomycis and micromonosporas.
Sources of drug
Mineral Source
Metallic and Non metallic sources.
Iron is used in treatment of iron deficiency anemia.
Mercurial salts are used in Syphilis.
Zinc is used as zinc supplement. Zinc oxide paste is used in wounds and in
eczema .
Iodine is antiseptic. Iodine supplements are also used.
Gold salts are used in the treatment of rheumatoid arthritis.
Miscellaneous Sources
Fluorine has antiseptic properties.
Borax has antiseptic properties as well.
Selenium as selenium sulphide is used in anti dandruff shampoos.
Petroleum is used in preparation of liquid paraffin
Marine Source
.
Sources of drug
•Marine Pharmacognosy is a sub-branch of Pharmacognosy, which is mainly
concerned with the naturally occurring substances of medicinal value from
marine.
•
•Generally the drugs are obtained from the marine species of bacteria,
virus, algae fungi, sponges etc.
•In the western medicine Agar, Alginic acid, Carrageenan,
Protaminesulphate, Spermaceti and Cod & halibut liver oils are the
marinemedicinal established products. Macroalgae or seaweeds have been
used as crude drugs in the treatment of Iodine deficiency states such as
Goiter etc
• Some seaweeds (algae growing in sea or rock below high water mark)
have also been utilized as sources of additional vitamins and in the
treatment of Anemia during pregnancy.
• Marine flora and fauna play significant role as a source of new molecular
entity
•
• During the past 30-40 years, Numerous novel compounds have been
isolated from marine organisms having biological activities such as
antibacterial ,antiviral, antitumour, antiparasitic, anticoagulants,
antimicrobial, antiinflammatory and cardiovascular compounds.
• of cough syrup.
Sources of drug
• Tissue Culture
• Tissue culture is invitro cultivation of plant cell or tissue under aseptic and
controlled environmental condition, in liquid and semi solid well defined
nutrient media for production of primary and secondary metabolites or to
regenerate whole plant.
• Eg. Ten times more production of anthraquinone derivatives from cassia
tora (6%) as compared to crude drug. The suspension culture of
Dioscorea deltoidea produce up to 1.5 % dry content of diosgenin. The
catharanthus roseus cell culture yielded 4 times more ajamalicene and
serpentine than the whole plant. (Shah B, 2010)
Sources of drug
Types of crude drugs
• 2 types exist depending on this type of
classification;
1. Organized drugs
2. Unorganized drugs
Organized drugs
These are drugs obtained from the direct parts of the plants and
containing cellular tissues
E.g. Rhizomes, barks, leaves, fruits, entire plants, hairs and fibers
Examples of organized drugs
Leaves– Digitalis, Eucalyptus, Mint, Senna, Spearmint, Squill, Tulsi, Vasaka, Coca,
Buchu, Hyoscyamus, Belladonna, Tea.
Barks–Cascara, Cassia, Cinchona, Cinnamon, Quillia.
Flowering parts– Clove, Pyrethrum, Chamomile.
•Fruits– Amla, Anise, Bitter Orange peel, Capsicum, Caraway, Cardamom,
Coriander, Dill, Fennel, Lemon peel, Star anise.
•Seeds– Bitter almond, Black Mustard, Cardamom, Colchicum, Ispaghula,,
Linseed, Nutmeg, Nux vomica.
•Roots and Rhizomes– Ashwagandha, Colchicum corm, Dioscorea, Garlic,
Ginger, Ginseng, Glycyrrhiza.
•Plants- Ephedra, Ergot, Bacopa.
•Hairs & fibres: cotton, jute, silk.
Unorganized drugs
These are drugs which are prepared from plants by some
intermediate physical process such as incision, drying or
extraction and not containing any cellular plant tissue
E.g. Aloe, honey, beeswax, essential oil
Differences between organized
and unorganized drugs
Organized Drugs Unorganized Drugs
These may be of plant or animal
origin.
These may be of plant, animal or
mineral origin.
These are direct part of plants or
animals.
These are the product of plant or
animals.
These have well defined cellular
structure.
These do not have well defined
cellular structure.
Generally identified by morphological
character.
Generally identified by organoleptic
properties.
Dried latex
The latex is a product contained in special secretory tissues of certain plants. It is
usually a white aqueous suspension, wherein microscopically small particles of oil
globules are suspended. These natural suspensions of milky consistency may
contain proteins, sugars, minerals and alkaloid salt in the true solution where gums,
starch and resins are in the suspended form. Examples of commercially important
lattices are opium, papain, ficin and gutta percha .
Dried Juice
The juices are obtained by fleshy leaves(aloe) or from stems of the tree(kino). In all
cases incisions are made to respective part of the plant and juice coming out is
collected and dried.
Dried Extract
The extract covered under crude drugs differ from galenical extracts. The extract of
pharmacognostic origin consisting of extracting the parts of the plants with water
followed by concentration , while pharmaceutical preparations known as extracts
are prepared by using alcoholic or hydroalcoholic solutions and adjusting the
Gums
Gums are translucent and amorphous substances produced by plants. Gums
are usually pathological products and are produced when the plant is growing
under unfavourable conditions or is injured. Thus they are the abnormal
products of plant metabolism. The gums are produced by the process known as
“Gummosis”.
Gums are soluble or partly soluble in water. They are insoluble in alcohol and
in most of the organic solvents. They form viscous adhesive solution with water
either by swelling or due to absorption. Aqueous solutions of gums are usually
leavorotatory. Gums are plant hydrocolloids and may be anionic or non ionic
polysaccharides. On hydrolysis(acidic hydrolysis or prolonged boiling with water),
gum yield sugar and uronic acids which form salts with calcium and magnesium.
The uronic acids are glucuronic acids are glucoronic acid, galactouronic acid or
aldobionic acid. Pharmaceutically important gums are gum acacia, tragacanth,
gum karaya, gum ghatti and guar gum
Mucilages
Mucilages are also plant products similar to gums and regarded to be the
normal products of plant metabolism. They are produced inside the cells of the
plant. Mucilages form slimy masses with water but do not dissolve. Mucilages
are esters of sulphuric acid wherein ester group is a polysaccharide complex.
Various examples along with methods of formation of mucilages are given
below:
From the cell wall of seed epidermis : Isabgol and linceed
From endodermis: Fenugreek
From leaf epidermis: Senna
From bark: Cinnamon, slipper elm
From special secretion cells : Squill
From algae: Agar and chondrus
• Oleo resin
When the natural plant resins are accompanied with the volatile oil in
homogeneous form are known as oleo resin. Canada balsams and copaiba are
suitable example of ole resin – do not compare these with prepared oleo
resins which are prepared by percolating drugs containing volatile oil and resin
together. The examples of this type are capsicum and podophyllum and ginger
oleo resin.
• Oleo-gum-resin
• These are the combinations of volatile oil, gum and resins. Sometimes they
also contain other substances like enzymes e.g. myrrh and asafoetida.
• The resins are found in different parts of plant or animal.
• External glands : Indian hemp
• In the heart wood of certain woods : guaiacum and red sanders
• Internal glands of plants : Male ferns
• Glands on surface of insects : shellac
B. Classification of crude drugs
CHAPTER-2
Classification of crude drugs may include;
 Alphabetical classification
 Morphological classification
 Pharmacological classification
 Chemical classification
 Taxonomic classification
 Chemo taxonomic classification
An ideal method of classification of crude drugs should be;
 Simple
 Easy to use
 Free from confusion
Alphabetical classification is the simplest way of classification
 Crude drugs are arranged in alphabetical order of their:
• Latin names
• English names
• Local/ vernacular names
 The following pharmacopoeia classify crude drugs according to this
system:
• British Pharmacopoeia
• British Herbal Pharmacopoeia
• Herbal Pharmacopoeia
• United States Pharmacopoeia and National Formulary
• British Pharmaceutical codex
• European Pharmacopoeia
 It is easy and quick to use
 There is no repetition of entries and is devoid of confusion.
 In this system location, tracing and addition of drug entries is easy
 There is no relationship between previous and successive drug
entries
 E.g. Acacia, Benzoin, Cinchona, Dill, Ergot, Fennel, Gentian,
Hyoscyamus, Ipecacuanha, Jalap, Kurchi, Liquorice, Mints,
Nuxvomica, Opium, Podophyllum, Quassia, Rauwolfia, Senna,
Vasaka, Wool fat, Yellow bees wax.
 Can not identify source of drug –plant, animal, mineral.
 In this system, the drugs are arranged according to the plant parts
or animal parts.
 Part of the plant is used as a drug e. g. leaves, roots, stem, fruit,
flower, rhizome.
 Convenient for practical study.
 Helpful in identification & detection of adulteration.
Disadvantage:
 No corelation between chemical constituents and therapeutic
action.
 Repetition of drugs.
 This involves grouping of drug according to their pharmacological
action.
 This is also referred to as therapeutic classification of drugs.
 Drugs like digitalis, squill and strophanthus having cardiotonic
action are grouped together irrespective of their parts used or their
phytoconstituents.
Pharmacological Action Drugs
Anticancer Vinca, Podophyllum, Taxus
Anti-inflammatory Colchicum, Turmeric
Antiamoebic Ipecac root, Kurchi bark
Antiasthmatic Ephedra, Lobelia
Anthelminthic Male fern, Quassia wood
Antispasmodic Datura, Hyoscyamus
Astringent Catechu
Analgesic Opium, poppy
Bitter tonic Quassia wood, Nux-vomica, Gentian
Carminatives Coriander, fennel, clove, peppermint
Purgatives Senna, Rhubarb
Expectorant Tulsi, Balsam of Tolu
Cardiotonic Digitalis, Squill, Strophanthus
ADVANTAGE
 This system of classification can be used for suggesting substitutes
of drugs if they are not available at a particular place or point of
time.
DISADVANTAGE
 Drugs having different action on the body gets classified separately
in more than one group that causes confusion.
 E.g. Cinchona is an antimalarial drug because of presence of
quinine but can be put under the group of drug affecting heart
because of antiarrythymic action of quinidine.
 The crude drugs are divided into different groups according to the
chemical nature of their relevant constituent.
 The chemical classification of drugs is dependent upon the
grouping of drugs with identical constituents.
1. Carbohydrates
 Gums - Acacia, Tragacanth, Guargum
 Mucilages - Plantago seed
 Others include Starch, Honey, Agar, Pectin, Cotton .
 Anthraquinone Glycosides - Aloe, Cascara, Rhubarb, Senna
 Saponins Glycosides - Quillaia, Arjuna, Glycyrrhiza
 Cyanogenetic Glycosides – Bitter almond, wild cherry bark.
 Isothiocyanate Glycosides - Mustard
 Cardiac Glycosides - Digitalis, Strophantus , squill
 Bitter Glycoside- Gentian, quassia, Chirata, Kalmegh.
 Tannins are astringent, bitter plant based poly hydroxy
phenols that either bind or precipitate proteins.
E.g.-pale catechue
Black catechue
Amla, Behra,Harde, Myrobalan etc.
 Monoterpenes and sesquiterpenes obtained from plants.
 Examples- Cinnamon, Fennel, Dill, Caraway, Coriander, Cardamom,
Orange peel, Mint, Clove
 Fixed oils – Castor, Olive, Almond, Shark liver oil
 Fats – Theobroma, Lanolin, Lard
 Waxes – Beeswax, Spermaceti , Wool fat.
 Nitrogenous substances of plant origin
 Pyridine and Piperidine – Lobelia, Nicotiana
 Tropane - Coca, Belladonna, Datura, Stramonium, Hyoscyamus,
Henbane
 Quinoline – Cinchona
 Isoquinoline – Opium, Ipecac, Calumba
 Indole – Ergot, Rauwolfia
 Amines – Ephedra
 Purine bases – Tea, coffee
 Complex mixture of resinol, resin acids, resitannols, resene etc.
 Eg. Colophony, podophyllum, jalap, capsicum, turmeric, asafoitida
etc.
d on an accepted system of botanical classification
 Natural relationship
 Grouped into:
• Kingdom- Plant
• Division-Angiosperm
• Class- Dicotylendons
• Order- Tubiflorae
• Family- Solanaceae
• Genus- Datura
• Species- metal
• Variety- fastuosa
Plant kingdom
Thallophyta
Algae, Fungi
Eg. Agar
Red algae
Pteridophyta
Derived name
from fern
Silent feature of
plants
Rise in pot as
ornamentals
eg,. Male fern
Gymnosperm
Gymno- naked
Sperm-seed
Plant body is
sporophytes and
differentiate
into root, stem
and leaves.
Eg. Ephedra
Angiosperm
Enclosed seed
Dicot
Two
cotylendon
s
Coriander
capsicum
Monocot
One
cotylendon
s
Colchicum
Maize
Allows for precise and ordered arrangement of drugs.
 Accommodates any drug without ambiguity
DISADVANTAGES
Can not recognize drug is organized or unorganize
 Chemo- nature of active constituents of drug
 Taxonomy- science deal with identification, nomenclature
and classification of plants.
 It involves study of chemical variation in different type of
plant and used of this information in classification of crude
drugs.
 Generally secondary metabolites are studied.
e.g.. Tropane alkaloids- solanaceae family
Isothiocynates- crucifareae
Anthocynidines- red, blue and other colors of flower.
DRUG PLANT SOURCE USE(S)
Vinblastine Catharanthus roseus Anticancer
Vinblastine Catharanthus roseus Anticancer
Reserpine Rauvolfia serpentina Antipsychoyic,
antihypertensive
Quinine Cinchona sp. Antimalarial,
amoebic dysentery
Pilocarpine Pilocarpus jaborandi Antiglucoma
Cocaine
Lidocaine, procain
Erythroxylum coca Topical anesthetic
DRUG PLANT SOURCE USE(S)
Codeine Morphine Papaver somniferum Antitussive, Narcotic
analgesic
Atropine Atropa belladonna Spasmolytic
Artemisinin Artemesia annua Antimalarial
Taxol Taxus baccata,
T. brevifolia
Breast and ovarian
cancer
Allicin Allium sativum Antifungal, amoebiasis
Podophyllotoxin
Etoposide, Teniposide
Podophyllum Anticancer
Adulteration and Quality control of crude drug
c
• Inferiority is a natural substandard condition (e.g. where a crop is taken whose
natural constituent is below the minimum standard for that particular drug)
which can be avoided by more careful selection of the plant material.
• Eg- minimum value of strychnine is 1.15%
• Less than 1.15% strychnine content - inferior
• The term 'adulteration' or debasement of an article covers a number of
conditions, which may be deliberate or accidental. Usually in crude
drugs, this practice includes substitution of the original crude drugs
partially or fully with other substances which is either free from or
inferior in therapeutic and chemical properties.
• DEFINATION- comes from Latin word which means mixing the original
drug material with other false material.
• Spoilage is a substandard condition produced by microbial or other pest
infestation, which makes a product unfit for consumption, which can be
avoided by careful attention to the drying, and storage conditions.
• Deterioration Due to age Less chemical component of Drug,also can be due to
high temp., light, improper storage condition, bacterial growth or intentional
extraction of the constituents and the sale of the residue as the original drugs.
• Admixture is the addition of one article to another through accident,
ignorance or carelessness e.g.
• During collection of root and rhizome, stones are collected.
• inclusion of soil on an underground organ or the co- collection of two similar
species.
• Sophistication intentional addition of other substance in original drug with
intent to defraud; such materials are carefully produced and may appear
at first sight to be genuine
• e.g. powder ginger may be diluted with starch with addition of little
coloring material ( turmeric ) to give the correct shade of yellow colour.
• Substitution is the addition of an entirely different subastance in
replacement of that which is required e.g. supply of cheap cottonseed oil
in place of olive oil.
• Addition of water in cow milk- adultration
• Replacement of goat milk in place of cow milk- substitution
• Non availability
• Ignorance and inadequate knowledge of collectors
• Regional/ linguistic nomenclature
• Ecotype, genotype, chemo-type variation
• Making profit
Different methods used for adulteration may be grouped as follows:
1.Substitution with Inferior Commercial Varieties
Due to morphological resemblance to the authentic drugs, different inferior
commercial varieties are used as adulterant which may or may not have any
chemical or therapeutic potential as that original natural drug.
E.g. Arabian Senna (Cassia angustifolia),dog Senna (Cassia obovata) and
avaram (Cassia auriculata) have been used to adulterate Senna (Cassia
senna);
-
TYPES OF ADULTERATION OR SUBSTITUTION OF HERBAL DRUGS:
TYPES OF ADULTERATION OR SUBSTITUTION OF HERBAL DRUGS:
TYPES OF ADULTERATION OR SUBSTITUTION OF HERBAL
DRUGS:
- Japanese ginger (Zingiber mioga), African and cochin ginger to adulterate
medicinal ginger (Zingiber officinale).
- Strychnous nux-blanda or S. potatorum in place of S. nux-vomica
- Capsicum minimum replaced by C.annuum.
- Gentian substituted by kutki
2. Adulteration by Artificially Manufactured Substitutes
• To provide the general form and appearance of various drugs, some
materials are artificially manufactured and are used as substitute of
the original one
• Generally this practice is for much costlier drugs.
•
E.g.
- artificial invert sugar for honey;
- paraffin wax after yellow coloration substituted for bees wax.
- Compressed chicory in place in place ofcoffee
- Properly cut and shaped baswood for Nutmeg
3. Substitution by Exhausted Drugs
Here the same plant material is mixed which is having no active medicinal
components as they have already been extracted out.
This practice is most common in case of volatile oil containing materials
like clove, fennel, coriander, caraway etc., where the dried exhausted
material resembles the same like original drug (similarly with drugs like
Cascara sagrada and ginger).
Sometimes when coloring matters have been extracted or removed during
exhaustion, the residue is re-colored with artificial dyes as is done with
saffron and red rose petals.
Exhausted ginger made bitter with aloes
4.Substitution by Superficially Similar but Cheaper Natural Substances
(Inferior drugs)
Usually here the adulterated product has no relation with the genuine
article, may or may not have any therapeutic or chemical component
desired,
Due to their morphological resemblance, they are marketed as adultrants.
- leaves of species - Ailanthus are substituted for belladonna, senna, mint etc.;
-Leaves of Phytolacca and Scopolia for belladona;
-Leaves of Xanthium for stramonium and dandelion for henbane;
-Indian dill with European dill or caraway etc.
Mother cloves and clovestalks are mixed with clove
-Beeswax is substituted by Japan wax.
5. Adulteration by Addition of Worthless Heavy Materials i.e. Harmful
adulterants
The wastes from market are collected and admixed with authentic drug.
This is particularly noticed for liquids or unorganised drugs.
E.g.
-A large mass of stone mixed with Liquorice root
-pieces of limestone are found in asafoetida
-
- lead shot has occurred in pieces of opium
-Pieces of ambered coloured glass in colophony
-White oil in coconut oil
-Cocoa butter mixed with stearin or paraffin
The addition of rodent faecal matter to cardamom seed is a very harmful
adulterant.
6.Addition of Synthetic Principles
Sometimes to fortify inferior natural products, synthetic principles are added
E.g. adding citral to oil of lemon and orange;
benzyl benzoate to balsam of Peru etc.
7. Usage of Vegetative Matter from the Same Plant
This is done by mixing adventitious matters, miniature plants growing
alongwith with medicinal plant in excessive amount or parts of plant other
than that which constitutes the drugs due to their resembling colour, odour
and in some cases constituents.
E.g. The lower plants like moss,liver warts and epiphytes growing in bark
portion are mixed with Cascara or Cinchona;
- stems of buchu are sometimes cut into short lengths and added to the drug.
- Stem portions are mixed alongwith leaf drugs like stramonium, lobelia and
senna.
8. Adulteration of Powders
Powdered forms are frequently found to be adulterated.
E.g.
-Dextrin in Ipecacuanha
-Powdered liquorice or gentian admixed with powdered olive stones
-Exhausted ginger powder in powdered colocynth or ginger
-Red sanders wood in capsicum
The powdered bark is frequently found to be adulterated with brick powder.
EFFECT OF USING ADULTERANTS
Effect of using adulteration
EFFECT OF USING ADULTERANTS
EFFECT OF USING ADULTERANTS
Adulterated or spurious raw materials may not possess the
therapeutically active constituents, or
even may be having some toxic compounds that may not only affect the
efficacy of the finished product but also cause deleterious effect on
human health.
•Reliability of the finished product is at national and international level
is affected and betraying the faith of people on the Indian System of
Medicine
DRUG- EVALUATION
Drug Evaluation means conformation of its identity and determination of its
quality and purity and detection of nature of adulteration.
It is necessary because
1) Biochemical variation in the drug
2) Deterioration due to treatment and storage
3) Substitution and adulteration, as a result of carelessness, ignorance or fraud.
Initially, crude drugs were identified by comparison only with the standard
description available.
Due to advancement, at present evaluation also includes method of estimating
active constituent present in the crude drug, in addition to its morphological and
microscopic analysis.
Its possible to perform physical evaluation which could be both qualitative and
quantitative nature
Different techniques involved are
1) Morphological and Organoleptic evaluation
2) Microscopic evaluation
3) Chemical evaluation
4) Physical Evaluation
5) Biological Evaluation
Standardization (Drug Evaluation) of Crude drugs
MORPHOLOGICAL EVALUATION
•Organoleptic evaluation means conclusions drawn from studies resulted due to
impressions on organs of senses.
Done for unorganized drug.
• Morphological evaluation means identification of drug by size, shape, specific
textute ( done for organized drug)
The study of form of crude drug is MORPHOLOGY, while description of the form is
MORPHOGRAPHY.
E.g.
-Fractured surface in cinchona, quillaia, cascara barks and quassia wood
(SHAPE) Nux vomica- disc
shape
Sarpgandha –wavy shape
(TASTE)Liquorice & stevia –
sweet taste
ODOUR- MENTHA & CLOVE-
AROMATIC
(COLOUR)Cardmom &
coriander if shade
dried- green colour
Sun dried- pale green
ORGANOLEPTIC EVALUATION
 Aromatic odor of Umbelliferous fruits
 -Sweet taste of Liquorice
 -Brown color of cinnamon
 -Odor and taste of spice-drugs like, asafoetida, black pepper, nutmeg,
caraway, cummin etc
MICROSCOPIC EVALUATION
• Allows more detailed examination of a drug BY USE OF MICROSCOPE
• Can be used to identify the organised drugs by their known histological
characters (cellular)
• Mostly used for qualitative evaluation of organised crude drugs in entire and
powdered forms .
CHEMOMICROSCOPY:
• For the effective results, various reagents or stains can be used to distinguish
cellular structure (chemo-microscopy)
 A drop of phloroglucinol and conc. HCL give red stain with lignin.
 Mucilage is stained pink with Ruthenium red
QUANTITATIVE MICROSCOPIC EVALUATIONS-
it involves a specific histological characters.
LEAF CONSTANT CONTAINS 5 PARAMETERS
1. STOMATAL NUMBER
2. STOMATAL INDEX
3. VEIN ISLET NUMBER
4. VEIN TERMINATION NUMBER
5. PALISADE RATIO
•1.LEAF CONSTANT : Stomatal number and index
- Why- to identify crude leafy drugs & to distinguish between some closely
related species.
- Evaluation and purity determination of crude leafy drug
- Eg- Indian senna & Alexandrian senna.
- i) Stomatal number – is average number of stomata per sq. mm of the
leaf.
- Example- Indian senna stmatal no. in upper epidermis 220-260, lower
epidermis-240-265
ii) Vein islet number – is defined as the number of vein-
islets per sq. mm of the leaf surface midway between midrib
and the margin. Levin in 1929 determined vein islet no.
Name of Plant Range
Adhatoda vasica 6-8
Datura stramonium 12-16
Digitalis purpurea 2.5-3.0
Eucalyptus globulus 8-13.5
Ocimum sanctum 19-23
2) Stomatal Index – it is the percentage proportion of number of stomata form from
the total no. of epidermis cell. Stomatal index are constant.
Calculated by using the following equation:
S.I. = [S / (E + S) ] X 100
Where, S.I.= Stomatal Index
S = No. of stomata per unit area
E = No. of epidermal cells in the same unit area
iii) Vein-termination number - is defined as the number of
veinlet terminations per sq. mm of the leaf surface midway
between midrib and margin.
Leaf constants or Diagnostic characters of leaves
i)Palisade ratio – is defined as average number of palisade cells beneath each
epidermal cell. It can be determined with powdered drugs also.
Table: Palisade ratios of various leaf drugs
Name of Plant Range
Adhatoda vasica 5.5-6.5
Datura stramonium 4.2-6.5
Digitalis purpurea 3.7-4.2
Eucalyptus globules 5.5-6.5 (upper)
Ocimum sanctum 02-3.5
STOMATA
It is a minute epidermal opening present on aerial parts of the plants, with followin
characteristics:
i) A central pore
ii) Two kidney shaped similar cells containing chloroplasts known as guard cells
and varying number of subsidiary (epidermal) cells covering the guard cells.
 Function of stomata is gaseous exchange and the secondary function
transpiration.
 It is not essential that each plant must have stomata.
EG-They are also present in stems (Ephedra), flowers (clove), and fruits (fennel).
 It is generally observed that stomata are abundantly present in dicot leaves.
 In some cases, they are present on the upper surface of leaves, while in others on
lower surface only (coca and cherry).
 In some, the stomata are present on both surfaces of the leaves (senna,
belladonna, datura etc.).
 The distribution of stomata between upper and lower epidermis in dicot leaves
shows great variation.
 Types of Stomata
•Depending upon the guard cells and arrangement of subsidiary cells, stomata are
divided into four types:
1. Moss type
2. Gymnospermous type
3. Gramineous type
4. Dicotyledonous type
ACCORDING TO ARRANGEMENT OF SUBSIDARY CELLS:
i) Paracytic or Rubiaceous stomata
This type of stomata comprises of two guard cells covered by two subsidiary
cells, the long axis of which are parallel to that of stoma
e.g. coca and senna leaves
ii) Diacytic or caryophyllaceous or cross-celled stomata
The guard cells are covered by two subsidiary cells, as in case of paracytic
stoma, but the arrangement of subsidiary cells on the guard cell is at right
angle to that of stoma,
e.g. Peppermint, spearmint and Vasaka
iii) Anisocytic or cruciferous or unequal celled stomata:
The guard cells are covered by three subsidiary cells, of which one is markedly
smaller than the other two,
e.g. Belladonna, datura and stramonium
iv) Anomocytic or ranunculaceous or irregular celled stomata:
Stoma is surrounded by varying number of subsidiary cells resembling other
epidermal cells
e.g. Digitalis, lobelia, buchu
IP recognise one more type of stomata,
v) Actinocytic or radiate celled stomata
Two guard cells are surrounded by a circle of radiating subsidiary cells.
TRICHOMES
• Trichomes are the tubular elongated or glandular outgrowth of the epidermal
cells .
• Trichomes are also called as plant hairs. trichomes consists of two parts root
and body.
• Ttrichomes present in most of plant parts and are function less but some
times perform secretory function.
• Depending up on the structure and the number of cells present in
trichomes,they are classified in to following.
1.Covering Trichomes or non glandular trichomes:
2.Glandular Trichomes: eg dhatura
3.Hydathode or special Trichomes:
Trichomes absent in coca, less in senna leaves.
TRICHOMES
QUANTITATIVE MICROSCOPY
Lycopodium spore method:
It is use to determination of percentage of foreign organic matter, mainly use
for powdered drugs. Developed by T.E WALLIS.
• It is used when especially chemical and other methods of evaluation of drugs
fails to determine quality.
• Lycopodium spores are very characterized in shape and appearance and
uniform in size (25μm) on avg., 94000 spores present/mg of lycopodium
powder.
• It consists of 1.well defined particles which may be counted. 2.Single layered
cells or tissues the area of which may be traced under suitable magnification
and actual area calculated 3.The objects of uniform thickness, the length of
which can be measured, and actual area calculated.
PHYSICAL EVALUATION
These are rarely constant for crude drug , but may help in evaluation with
reference to crude drug.
1. Moisture content
2. Specific gravity
3. Density
4. Optic rotation
5. Refractive index
6. Melting point
7. Viscosity
1. Moisture content is responsible for the decomposition of crude drug due
to chemical change or microbial attack. It is necessary to determine and
control the moisture content of crude drug . It is determined by heating
the drug at 105°C in an oven to a constant weight. Moisture content of is
5% w/w.
2. Viscosity:
Resistance of the fluid to flow .
Viscosity of a liquid is constant at given temperature .
Hence it is used as a means of standardizing liquid drugs.
PYROXYLIN kinematic viscosity is 1100-2450 centistokes .
LIQUID PARAFFIN kinematic viscosity is 64 centistokes at 37.8°C.
3. Melting Point:
It is one of the parameter to judge the purity of crude drug. In case of
pure chemical or phytochemicals , melting point are very sharp and
constant . Since the crude drug from animals and plants origin contain
the mixed chemicals . Purity of crude drug can be determined by their
melting points. Eg. COLOPHONY 75-85°C, BEES WAX 62-65°C, WOOL
FAT 34-44°C COCOA BUTTER 30-33°C.
Certain drugs fluoresce when the cut surface or the powder is exposed
to
Certain drugs fluoresce when the cut surface or the powder is exposed to
ultraviolet radiation , And it is useful in the identification of those drugs .
Indian and Chinese Rhubarb are very difficult to distinguish and is very difficult
form , but examination in ultraviolet light gives such marked differences in
florescence that the varieties can be easily distinguish from each other .
4. Solubility:
The number of ml of solvent required to dissolve 1g of drug . The presence of
adulteration in a drug could be indicated by solubility studies. Eg. Balsam of
peru is soluble in chloral hydrate solution, Colophony is freely soluble in light
petroleum, Asafoetida is soluble in carbon disulphide., Alkaloidal bases are
soluble in chloroform.
The following are the most important parameter with respect to crude drug.
Ash Value:
The residue remaining after incineration is the ash content of the drug
(inorganic salts of carbonates , phosphates , silicates of sodium , potassium,
calcium and magnesium ) is known as ash content.
Ash value is the criterion to judge the identity and purity of crude drug. It is
also useful for detecting low grade products, exhausted drug and drugs from
earthy matter .
There are three types of Ash value:
Total Ash
Acid Insoluble Ash
Water soluble Ash
The total ash includes both “physiological
ash”, which is derived from the plant tissue
itself, and “non-physiological” ash, which is
the residue of the extraneous matter (e.g.
sand and soil) adhering to the plant surface.
Acid-insoluble ash is the residue obtained
after boiling the total ash with dilute
hydrochloric acid, and igniting the remaining
insoluble matter. This measures the amount
of silica present, especially as sand and
siliceous earth.
Water-soluble ash is the difference in weight
between the total ash and the residue after
treatment of the total ash with water.
Extractive Value
• It is used to detect either material exhausted by water or not (tea leaves ,
ginger rhizomes) The extract obtained by exhausting crude drugs with different
solvents are approximate measures of their chemical constituents .
• Various solvents are used according to the type of the constituents to be
analyzed .
• Useful for the evaluation especially when the constituents of the drugs can
not be readily estimated by any other means .
• It also helps to indicate the nature of chemical constituents present in the
drug.
• Also helps in the identification of adulteration of drug .
Solubility of drug is also detected .
• There are three types of extractive value.
Water soluble extractive value
Alcohol soluble extractive value
Ether soluble extractive value.
1. Water soluble Extractive
It is applied for the drugs which contain water soluble constituents such as
tannins , sugar , plant acids and mucilage.
2. Alcohol soluble Extarctive
It is applied for drugs which contain alcohol soluble constituents such as
tannins, resins and alkaloids.
3. Ether soluble Extractives:
It is applied for the extraction of volatile oils , fixed oils and resins .
Volatile ether soluble extractive value
Non volatile ether soluble extractive value.
.
Foreign Organic matter :
The parts of the organ or organs other than those named in the definition and
description of the drug are defined as foreign organic matter. The maximum limit for
the foreign organic matter is defined in the monograph of crude drug. If it exceedsthe
limits, deterioration in quality of the drug takes place.
Swelling factor:
Significances : Useful in the evaluation of crude drugs containing mucilage
Useful for the detection of purity of the crude drug
Determination : 1. Transfer 1 gm of the seeds to a 25ml stoppered cylinder
2. Fill up to the 20ml mark on the cylinder with water. Agitate gently and
occasionally during 24 hours and allowed to stand
3.Measure the volume occupied by the swollen seeds.
and occasionally during 24 hours and allowed to stand
3.Measure the volume occupied by the swollen seeds.
Spectrophotometric methods:
i) UV- Ultra violet /visible spectroscopy
ii)IR-Infra Red spectroscopy
iii) Fluorescence analysis
iv) NMR-nuclear magnetic resonance spectroscopy
v) MS-Mass spectroscopy
vi) X-ray diffraction
vii) RIA-radio immuno assay.
Above mentioned all spectrphotometric evaluation techniques are useful for
structure elucidation of Phytoconstituents.
Chromatographic techniques:
TLC-Thin layer chromatography
Principle :Adsorption
Adsorbent silica gel G/C coated to a thickness of minutes and used.
After development of chromatography spots are revealed by spraying with
suitable detecting agent
TLC is useful to analyse Alkaloids, Glycosides like all bio- constituents.
The Rf value vary depend on the nature of substance, composition of solvent
and impurities.
Rf value =
TLC/HPTLC are micro analytical techniques used for determination of natural
products Advantages : simple in operation and rapid Chromatographic
techniques.
CHEMICAL EVALUATION
Determination of the active constituent in a drug by chemical tests is referred to
as chemical evaluation.
The following are various methods of chemical evaluation
1. Instrumental methods
2. Chemical tests
3. Individual constituent chemical tests
4. Micro chemical tests
1. Instrumental methods: They make use of various instruments for evaluation like
colorimetry, flourimetry, spectrophotometry etc.
2. Chemical constants tests: These are like acid value, iodine value and ester value
etc are used for the identification of fixed oils and fats.
3. Individual chemical tests: These are the tests which are used for identifying
particular drugs.
4. Micro-chemical tests: These are the tests which are carried on slides. Example:
Euginol in clove oil is precipitated as potassium euginate crystals.
Method for chemical evaluation
Extract obtained using petroleum ether, chloroform, ethanol and water was
prepared using the respective solvent. These extracts along with positive and
negative controls were tested for the presence of active phytochemicals viz:
tannins, alkaloids, phytosterols, triterpenoids, falvonoids, cardiac glycosides,
anthroquinone glycosides, saponins, carbohydrates, proteins, amino acids and
fixed oils & fats following standard methods
Tannins
1. Ferric chloride Test: Added a few drops of 5% ferric chloride solution to 2 ml of
the test solution. Formation of blue color indicated the presence of hydrolysable
tannins.
2. Gelatin Test: Added five drops of 1% gelatin containing 10% sodium chloride to 1
ml of the test solution. Formation of white precipitates confirmed the test.
Alkaloids
Approximately 50 mg of extract was dissolved in 5 ml of distilled water. Further 2M
hydrochloric acid was added until an acid reaction occurred and filtered. The
filtrate was tested for the presence of alkaloids as detailed below
1.Dragendorff’s Test: To 2 ml of the filtrate was added 1 ml of Dragendorff’s
reagent. Formation of orange or reddish brown precipitate indicated the test as
positive.
2.Mayer’s Test: To 1 ml of test solution or filtrate was added a drop or two of the
Mayer’s reagent. white or a creamy precipitate confirmed the test as positive.
3.Hager’s Test: To 1 ml of test solution or filtrate, a drop or two of Hager’s reagent
formation of yellow precipitate indicated the test as positive.
4.Wagner Test: Two drops of Wagner’s reagent was added to 1ml of the test
solution. The formation of yellow or brown precipitate confirmed the test as
positive for alkaloids.
Phytosterols
1. Liebermann-Burchard’s Test: The extract (2 mg) was dissolved in 2 ml of acetic
anhydride, heated to boiling, cooled and then 1 ml of concentrated sulfuric acid was
added. A brown ring formation at the junction and the turning of the upper layer to
dark green color confirmed the test for the presence of phytosterols.
Triterpenoids
Salkowski Test: Approximately 2 mg of dry extract was shaken with 1 ml of
chloroform and a few drops of concentrated sulfuric acid were added.A red brown
color formed at the interface indicated the test as positive for triterpenoids.
Saponins
1.Foam Test: 5 ml of the test solution taken in a test tube was shaken well for five
minutes. Formation of stable foam confirmed the test.
2. Olive oil test: - Added a few drops of olive oil to 2ml of the test solution and
shaken well. The formation of a soluble emulsion confirmed the test.
Flavonoids
1.Shinoda test: A few magnesium turnings and 5 drops of concentrated hydrochloric
acid was added drop wise to 1 ml of test solution. A pink, scarlet, crimson red or
occasionally green to blue color appeared after few minutes confirmed the test.
2. Alkaline reagent test: Addition of 5 drops of 5% sodium hydroxide to 1 ml of the
test solution resulted an increase in the intensity of the yellow color which became
colorless on addition of a few drops of 2 M hydrochloric acid which indicated the
presence of falvonoids.
3.Lead acetate test: A few drops of 10% lead acetate added to 1ml of the test
solution resulted in the formation of yellow precipitate confirmed the presence of
falvonoids.
Cardiac glycosides
1.Keller -Killiani test: Added 0.4 ml of glacial acetic acid and a few drops of 5% ferric
chloride solution to a little of dry extract. Further 0.5 ml of concentrated sulfuric acid
was added .The formation of blue color in acetic acid layer confirmed the test..
Carbohydrates
1.Molisch’s test: To 1 ml of test solution added a few drops of 1 % alpha-napthol and
2-3 ml concentrated sulfuric acid. The reddish violet or purple ring formed at the
junction of two liquids confirmed the test.
2. Barfoed’s test: 2ml of reagent was added to 2 ml of the test solution, mixed & kept
a in boiling water bath for 1 min. Red precipitate formed indicates the presence of
monosaccharide's.
3. Seliwanoffs test: To 3 ml of Seliwanoffs reagent was added to 1 ml of the test
sample and heated on a water bath for one minute. The formation of rose red color
confirmed carbohydrates
4. Fehlings test: Dissolved 2 mg dry extract in 1 ml of distilled water and added 1ml of
Fehling’s(A+B) solution, shooked and heated on a water bath for 10 minutes. The
brick red precipitate formed confirmed the test.
Anthraquinone glycosides
Hydroxyanthraquinone Test: To 1 ml of the extract, added a few drops of 10%
potassium hydroxide solution. The Formation of red color confirmed the test.
.
Test for proteins:
Biuret test: To 2 ml of the test solution added 5 drops of 1% copper sulphate solution
and 2 ml of 10% NaOH .Mix thoroughly. Formation of purple or violet color confirmed
proteins.
Test for amino acids
1. Millon’s test: Added 5 drops of millons reagent to 1 ml of test solution and heated
on a water bath for 10 min, cooled and added 1% sodium nitrite solution.
Appearance of red color confirmed the test.
Fats and fixed oils :
To 5 drops of the sample was added 1 ml of 1% copper sulphate solution and a few
drops of 10% sodium hydroxide. The formation of a clear blue solution confirmed the
test.
BIOLOGICAL EVALUATION
• It is employed when the drug cannot be evaluated satisfactorily by chemical and
physical methods.
• In this method, the response produced by the test drug on a living system is
compared with that of the stranded preparation.
• Such an activity is represented in units as International Units (I.U).
Dose is termed as International units IU eg. Digitalis 1IU=76mg of standard ,Vit-A
1IU=0.344 of standard , Vit-D 1IU=0.025of standard.
Indication of Biological Evaluation
• When the chemical nature of the drug is not known but is has an biological action.
• When chemical methods are not available.
• When the quantity of the drug is small and so it cannot be evaluated chemically.
• Drugs which have different chemical composition but same biological activity.
Example: Cardiac glycosides are evaluated by this method on cats, frogs or pigeons.
SIGNIFICANCE
1.The method is generally used when standardization is not done satisfactory by
chemical or physical methods
2.When the quantity of the drug /sample are very less then the drugs are evaluated
by biological methods.
3.These methods are performed on living animals, isolating living organ and tissue,
animal preparation, and micro-organism ( Bioassay).
METHODS OF STUDIES
1)Toxic----animals are used
2)Symptomatic-----animals are used
3)Tissue-------isolated tissue is used
To estimate potency of drug ,With entire animal or with tissue, To conform
therapeutic activity.
Evaluation of Hypoglycemic activity deficiency of glucose in the bloodstream.
• Traditional Diabetic drugs -Momordica charantaka,Fenu greek,Gudmar.
Diabetis is induced in animals by Alloxan & Streptazocin.
• Alloxan cause necrosis of pancreatic islet-B cells which shows 180-250mg/ml fasting
blood glucose levels
• Streptazocin cause formation of streptomycin they produce cytotoxic nitrourcido
glucopyranose which cause diabetes
• ANIMALS USED: Rabbits,Rats,Mice—(4 to 7 days) Dose :rats--80mg/kg ,mice—
150mg/kg of streptazocin single oral injection 140-180 mg/kg of alloxan for rabbits at
marginal ear vein for 7 days for rats and mice intraperitoneally 2 days Insulin levels
are noted by tests like RIA.ELISA.
•RIA-detects antibody antigen reaction by using radioisotopes.
•ELISA-detects antibody antigen reaction by using enzyme.
Reference
WHO guidelines for assessing quality of herbal medicines By World Health
Organization
THANK YOU
www.paruluniversity.ac.in

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chapter -1 pharmacognosy bpharm 4th sem cognosy

  • 1. Pharmacognosy & Phytochemistry I BP405T-1 Ms. Sapna kumari Assistant Professor School of Pharmacy
  • 2. Introduction to Pharmacognosy Classification of drugs Quality Control of Drugs of Natural origin CHAPTER-1
  • 3. Definition, History, scope and development of Pharmacognosy A
  • 4. Definition • Study of crude drugs obtained from plant, animals and minerals and study of their active constituents. • Derived from Latin term • Pharmakon- A drug • Gignosco- To Acquire knowledge of • Pharmacognosy word first used as Pharmacognosis. • Term pharmacognosy first used by John .Adam .Schmidt, A Austrian Physicist and he used the term Pharmacognosist in 1811 in his work Lehrbuch der Materia Medica“. • After J. A. Schmidt, C.A Seydler used the term Pharmacognosis in his work Anlactica Pharmacognostica in 1815.
  • 5. WHAT IS PHARMACOGNOSY? • Pharmacognosy is the study of crude drugs from natural sources including information regarding History, Distribution, Cultivation & collection Sensory, physical and physicochemical characterization Structural features (Phytochemical Analysis) Medicinal uses, Processing of crude drug for commercial market Storage Preservation of drug.
  • 6. AD (ANNO DOMINI) In the year of lord BC (BEFORE CHRIST) 0 BIRTH OF JESUS CHRIST  Concept of AD and BC was given by POP DIONYSIUS, he was a Christian. Before the year of Christ
  • 7. History In India medicinal properties of plants described in Rigveda and Ayurveda (3500- 1500 B.C.) Charaka Samhita – plants Sushruta Samhita-surgery Papyrus Eber (1500 B.C.)- it is an oldest written document of egypt it contains 876 formulations and 700 crude drugs. In china medicinal plants had been used since 5000 B.C. Egyptians (Ebers papyrus, 1550 BC)
  • 8. History Hippocrates –Father of medicine (460-360 BC) • He was a Greek scientist and has described human anatomy and physiology. • He has also describe collection, identification, uses, property, of around 300 drugs.
  • 9. • Aristotle- Father of biology(384-322 BC) • Student of Pluto & known for his writing on animal kingdom. • Theophrastus - Father of botany (384-322 BC) • Student of Aristotle wrote about plant kingdom. • Galen- Described method of preparations of plant and animal drugs known as “Galenicals”
  • 10. History • Dioscorides • “De Materia Medica” (600 medicinal plants) • Described some medicinal plants like Belladona, Ergot, Opium, Colchicum etc.
  • 11. History • Swede linnaeus –Classified plants and introduced binomial system for naming plants. • It is a system used to name species • Eg- genus-homo, species-sepiens • To ensure that scientific name of an organism is same all over the world. • Classification further developed by Bentham & Hooker- Monocotyledon, dicotyledon & gymnosperm. • Eicher & Engler. • Mendel – Study on hybrids. • Paracelus – Developed mineral salts. • Le’mary- Importance of extraction method and alcohol as extractant. • Willium withering – Published some of medicinal properties of fox glove leaves. • Stass & otto- Developed extraction process for alkaloids.
  • 12. Development • 1934-1960 • Development of modern pharmacognosy with integration of chemistry, biotechnology, analysis, cology etc. • Around 75 substance from plant were isolated and structures were determined along with pharmacological studies. 1928 Penicillin Antibiotic 1952 Reserpine Hypotensive and tranquilizer( drug reduce reduce tension) 1955 Vincrisitin and vinblastin Anticancer Blood and lymphcancer 1960 Diosgenin Preparation of sterodal hormones
  • 13. Development • 1960-1970 • Antibiotics like • Streptomycin, • Chloramphenicol, • Tetracycline , • Gresiofulvin etc isolated so called antibiotic age. • Phytoconstituents from • Digitalis • Senna • Aloe • Belladona • Ergot were isolated and their clinical used were established
  • 14. Development • SAR • Semi synthetic preparations • E.g. Ergometrin Methyl Ergometrin Better oxitocic activity • Study of biosynthetic pathways • Shikimmic acid & other biosynthetic path way Constituents Acivity Responsible group Reserpine Hypotensive Trimethoxy benzoic acid Tubocurarine Muscle relaxant Quaternary ammonium group
  • 15. Development • Natural molecule as model for synthesis of new drug • E.g. During world war –II Quinine as an antimalarial drug was short in supply so similar type of compound chloroquine was isolated • Cocaine to lignocaine, xylocaine prepared as local analgesic • Silicin to salicylic acid (Aspirin) prepared which is used as Analgesic, Anti thrombolytic and Anti inflammatory agent.
  • 16. Development • 1970-1990 Phytoconstituents Plant Name Activity Podophllotoxin (Etoposide, Teniposide) Podophyllum Lung and testicular cancer Silymarin Milk thistle Liver disease Taxol Taxus Ovarian, Breast and lung cancer Guggulosterol Guggul Arthritis, Hypercholesteremia Ginkolide Ginko Increase blood circulation in brain Ginsenoside Ginseng Tonic
  • 17. Development • 1990 onwards  Herbal drug demand increase due to less toxicity, side effect and cost.  To control quality WHO published guideline and prepared monograph.  Gov. Of India published Ayurvedic pharmacopoeia part I &II.  Indian drug manufacturers published 3 volume of Indian herbal pharmacopoeia.  USP 2010- 18 plants  IP 2009 - 27 plants  TKDL – Traditional knowledge digital library  Prepared by india from 2 feb 2009 to protect traditional knowledge.  7 crore TKDL project took 9 year to enlist 2 lakh formulations from Ayurveda Siddha and unani.  Available in english, German, Japaniese, french and spanish.
  • 18. Scope • Identification of proper drug by morphology and microscopy. • Quality of herbs known by evaluation. • Good quality of drug obtained by proper cultivation and harvesting, drying & storing at proper time. • Cultivation of drug which are in demand and earn by selling it. • Active constituents are isolated from plants and can be marked to earn.
  • 19. • Pharmacognosy is an important link between Pharmacology and Medicinal chemistry. • Crude drugs also provides the intermediate precursor for synthesis of new drug. • Better active constituent’s semisynthetic derivatives can be prepared. • Eg. Ergotamine to Methyl Ergotamine • Synthetic molecule based on plant active constituent provide better activity. Eg. Quinine –Chloroquine (Antimalarial activity) Vasicine – Bromohexine (Bronchodilator) Scope
  • 20. • we can obtained new medicine for new diseases like AIDS, Cancer etc. • Plant tissue culture and genetic engineering gives more advance natural products. • Natural products as Pharmaceutical aid. Accacia & tragacanth – Binding, emulsifying & suspending agent. Carbohydrates- Sweetening agent Volatile oil – Flavoring agent Fixed oil- Ointment base, emolient Starch- Disintigrating agent Piperine- Bioavaibility enhancer. Scope
  • 22. Exploration of medicinal plant in ancient time • Trial and error- try and discard until its serves the purpose • While searching for food- herbs and species are identified. • Signature of nature- similarity between human organ and plant part • Zoo pharmacognosy-animal self medication power • Accidental discovery- cinchona,penicillin
  • 23. • Drugs are substances that are used or intended to be used in the diagnosis, prevention, treatment or cure of diseases. In early times, these substances were derived from natural sources, of which plants took up the major share. With the introduction of technology, most drugs today are manufactured synthetically in the laboratory. The major sources of drugs can be grouped into the following 1. Plant source 2. Animal source 3. Microbial source 4. Mineral Source 5. Marine source 6. Tissue culture Sources of drug
  • 24. Plant Source Plant source is the oldest source of drugs. Most of the drugs in ancient times were derived from plants. Almost all parts of the plants are used i.e. leaves, stem, bark, fruits and roots. Leaves: a. The leaves of Digitalis Purpurea are the source of Digitoxin and Digoxin, which are cardiac glycosides. b. Leaves of Eucalyptus give oil of Eucalyptus, which is important component Sources of drug
  • 25. Plant Source c. Tobacco leaves give nicotine. d. Atropa belladonna gives atropine. Flowers: Poppy papaver somniferum gives morphine (opoid) Vinca rosea gives vincristine and vinblastine Rose gives rose water used as tonic. Photo of Papaver somniferum by Evelyn Simak Fruits: Senna pod gives anthracine, which is a purgative (used in constipation) Calabar beans give physostigmine, which is cholinomimetic agent. of cough syrup. Sources of drug
  • 26. • Seeds: • Seeds of Nux Vomica give strychnine, which is a CNS stimulant. • Castor oil seeds give castor oil. • Calabar beans give Physostigmine, which is a cholinomimetic drug. • Roots: • Ipecacuanha root gives Emetine, used to induce vomiting as in accidental poisoning. It also has amoebicidal properties. • Rauwolfia serpentina gives reserpine, a hypotensive agent. • Reserpine was used for hypertension treatment. • syrup. • Bark: • Cinchona bark gives quinine and quinidine, which are antimalarial drugs. Quinidine also has antiarrythmic properties. • Atropa belladonna gives atropine, which is anticholinergic. • Hyoscyamus Niger gives Hyosine, which is also anticholinergic.
  • 27. Sources of drug Animal source • Drugs obtained from animals sources are whole animals, glandular products (thyroid organ), liver extract, polypeptide venoms, non-peptide toxins, etc. Fish liver oil, musk, beeswax, hormones, enzymes, and antitoxins sera are the products obtained from animal sources (kokate). A large number of other natural products from animal sources are used as pharmaceutic excipient and others are used as important drugs or as nutritional supplements. The examples are as given below (Annamalaiuniversity): • Pancreas is a source of Insulin, used in treatment of Diabetes. • Urine of pregnant women gives human chorionic gonadotropin (hCG) used for the treatment of infertility. • Sheep thyroid is a source of thyroxin, used in hypertension.
  • 28. Sources of drug •Cod liver is used as a source of vitamin A and D. •Anterior pituitary is a source of pituitary gonadotropins, used in treatment of infertility. •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. Nowadays better drugs have replaced them. •cough syrup.
  • 29. Sources of drug Microbial Source Many life-saving drugs are obtained from microbes, Penicillium notatum is a fungus which gives penicillin. Actinobacteria give Streptomycin. Aminoglycosides such as gentamicin and tobramycin are obtained from streptomycis and micromonosporas.
  • 30. Sources of drug Mineral Source Metallic and Non metallic sources. Iron is used in treatment of iron deficiency anemia. Mercurial salts are used in Syphilis. Zinc is used as zinc supplement. Zinc oxide paste is used in wounds and in eczema . Iodine is antiseptic. Iodine supplements are also used. Gold salts are used in the treatment of rheumatoid arthritis. Miscellaneous Sources Fluorine has antiseptic properties. Borax has antiseptic properties as well. Selenium as selenium sulphide is used in anti dandruff shampoos. Petroleum is used in preparation of liquid paraffin
  • 31. Marine Source . Sources of drug •Marine Pharmacognosy is a sub-branch of Pharmacognosy, which is mainly concerned with the naturally occurring substances of medicinal value from marine. • •Generally the drugs are obtained from the marine species of bacteria, virus, algae fungi, sponges etc. •In the western medicine Agar, Alginic acid, Carrageenan, Protaminesulphate, Spermaceti and Cod & halibut liver oils are the marinemedicinal established products. Macroalgae or seaweeds have been used as crude drugs in the treatment of Iodine deficiency states such as Goiter etc
  • 32. • Some seaweeds (algae growing in sea or rock below high water mark) have also been utilized as sources of additional vitamins and in the treatment of Anemia during pregnancy. • Marine flora and fauna play significant role as a source of new molecular entity • • During the past 30-40 years, Numerous novel compounds have been isolated from marine organisms having biological activities such as antibacterial ,antiviral, antitumour, antiparasitic, anticoagulants, antimicrobial, antiinflammatory and cardiovascular compounds. • of cough syrup. Sources of drug
  • 33. • Tissue Culture • Tissue culture is invitro cultivation of plant cell or tissue under aseptic and controlled environmental condition, in liquid and semi solid well defined nutrient media for production of primary and secondary metabolites or to regenerate whole plant. • Eg. Ten times more production of anthraquinone derivatives from cassia tora (6%) as compared to crude drug. The suspension culture of Dioscorea deltoidea produce up to 1.5 % dry content of diosgenin. The catharanthus roseus cell culture yielded 4 times more ajamalicene and serpentine than the whole plant. (Shah B, 2010) Sources of drug
  • 34. Types of crude drugs • 2 types exist depending on this type of classification; 1. Organized drugs 2. Unorganized drugs
  • 35. Organized drugs These are drugs obtained from the direct parts of the plants and containing cellular tissues E.g. Rhizomes, barks, leaves, fruits, entire plants, hairs and fibers
  • 36. Examples of organized drugs Leaves– Digitalis, Eucalyptus, Mint, Senna, Spearmint, Squill, Tulsi, Vasaka, Coca, Buchu, Hyoscyamus, Belladonna, Tea. Barks–Cascara, Cassia, Cinchona, Cinnamon, Quillia. Flowering parts– Clove, Pyrethrum, Chamomile.
  • 37. •Fruits– Amla, Anise, Bitter Orange peel, Capsicum, Caraway, Cardamom, Coriander, Dill, Fennel, Lemon peel, Star anise. •Seeds– Bitter almond, Black Mustard, Cardamom, Colchicum, Ispaghula,, Linseed, Nutmeg, Nux vomica. •Roots and Rhizomes– Ashwagandha, Colchicum corm, Dioscorea, Garlic, Ginger, Ginseng, Glycyrrhiza. •Plants- Ephedra, Ergot, Bacopa. •Hairs & fibres: cotton, jute, silk.
  • 38. Unorganized drugs These are drugs which are prepared from plants by some intermediate physical process such as incision, drying or extraction and not containing any cellular plant tissue E.g. Aloe, honey, beeswax, essential oil
  • 39. Differences between organized and unorganized drugs Organized Drugs Unorganized Drugs These may be of plant or animal origin. These may be of plant, animal or mineral origin. These are direct part of plants or animals. These are the product of plant or animals. These have well defined cellular structure. These do not have well defined cellular structure. Generally identified by morphological character. Generally identified by organoleptic properties.
  • 40. Dried latex The latex is a product contained in special secretory tissues of certain plants. It is usually a white aqueous suspension, wherein microscopically small particles of oil globules are suspended. These natural suspensions of milky consistency may contain proteins, sugars, minerals and alkaloid salt in the true solution where gums, starch and resins are in the suspended form. Examples of commercially important lattices are opium, papain, ficin and gutta percha . Dried Juice The juices are obtained by fleshy leaves(aloe) or from stems of the tree(kino). In all cases incisions are made to respective part of the plant and juice coming out is collected and dried. Dried Extract The extract covered under crude drugs differ from galenical extracts. The extract of pharmacognostic origin consisting of extracting the parts of the plants with water followed by concentration , while pharmaceutical preparations known as extracts are prepared by using alcoholic or hydroalcoholic solutions and adjusting the
  • 41. Gums Gums are translucent and amorphous substances produced by plants. Gums are usually pathological products and are produced when the plant is growing under unfavourable conditions or is injured. Thus they are the abnormal products of plant metabolism. The gums are produced by the process known as “Gummosis”. Gums are soluble or partly soluble in water. They are insoluble in alcohol and in most of the organic solvents. They form viscous adhesive solution with water either by swelling or due to absorption. Aqueous solutions of gums are usually leavorotatory. Gums are plant hydrocolloids and may be anionic or non ionic polysaccharides. On hydrolysis(acidic hydrolysis or prolonged boiling with water), gum yield sugar and uronic acids which form salts with calcium and magnesium. The uronic acids are glucuronic acids are glucoronic acid, galactouronic acid or aldobionic acid. Pharmaceutically important gums are gum acacia, tragacanth, gum karaya, gum ghatti and guar gum
  • 42. Mucilages Mucilages are also plant products similar to gums and regarded to be the normal products of plant metabolism. They are produced inside the cells of the plant. Mucilages form slimy masses with water but do not dissolve. Mucilages are esters of sulphuric acid wherein ester group is a polysaccharide complex. Various examples along with methods of formation of mucilages are given below: From the cell wall of seed epidermis : Isabgol and linceed From endodermis: Fenugreek From leaf epidermis: Senna From bark: Cinnamon, slipper elm From special secretion cells : Squill From algae: Agar and chondrus
  • 43. • Oleo resin When the natural plant resins are accompanied with the volatile oil in homogeneous form are known as oleo resin. Canada balsams and copaiba are suitable example of ole resin – do not compare these with prepared oleo resins which are prepared by percolating drugs containing volatile oil and resin together. The examples of this type are capsicum and podophyllum and ginger oleo resin. • Oleo-gum-resin • These are the combinations of volatile oil, gum and resins. Sometimes they also contain other substances like enzymes e.g. myrrh and asafoetida. • The resins are found in different parts of plant or animal. • External glands : Indian hemp • In the heart wood of certain woods : guaiacum and red sanders • Internal glands of plants : Male ferns • Glands on surface of insects : shellac
  • 44. B. Classification of crude drugs CHAPTER-2
  • 45. Classification of crude drugs may include;  Alphabetical classification  Morphological classification  Pharmacological classification  Chemical classification  Taxonomic classification  Chemo taxonomic classification
  • 46. An ideal method of classification of crude drugs should be;  Simple  Easy to use  Free from confusion
  • 47. Alphabetical classification is the simplest way of classification  Crude drugs are arranged in alphabetical order of their: • Latin names • English names • Local/ vernacular names
  • 48.  The following pharmacopoeia classify crude drugs according to this system: • British Pharmacopoeia • British Herbal Pharmacopoeia • Herbal Pharmacopoeia • United States Pharmacopoeia and National Formulary • British Pharmaceutical codex • European Pharmacopoeia
  • 49.  It is easy and quick to use  There is no repetition of entries and is devoid of confusion.  In this system location, tracing and addition of drug entries is easy
  • 50.  There is no relationship between previous and successive drug entries  E.g. Acacia, Benzoin, Cinchona, Dill, Ergot, Fennel, Gentian, Hyoscyamus, Ipecacuanha, Jalap, Kurchi, Liquorice, Mints, Nuxvomica, Opium, Podophyllum, Quassia, Rauwolfia, Senna, Vasaka, Wool fat, Yellow bees wax.  Can not identify source of drug –plant, animal, mineral.
  • 51.  In this system, the drugs are arranged according to the plant parts or animal parts.  Part of the plant is used as a drug e. g. leaves, roots, stem, fruit, flower, rhizome.
  • 52.  Convenient for practical study.  Helpful in identification & detection of adulteration. Disadvantage:  No corelation between chemical constituents and therapeutic action.  Repetition of drugs.
  • 53.  This involves grouping of drug according to their pharmacological action.  This is also referred to as therapeutic classification of drugs.  Drugs like digitalis, squill and strophanthus having cardiotonic action are grouped together irrespective of their parts used or their phytoconstituents.
  • 54. Pharmacological Action Drugs Anticancer Vinca, Podophyllum, Taxus Anti-inflammatory Colchicum, Turmeric Antiamoebic Ipecac root, Kurchi bark Antiasthmatic Ephedra, Lobelia Anthelminthic Male fern, Quassia wood Antispasmodic Datura, Hyoscyamus Astringent Catechu Analgesic Opium, poppy Bitter tonic Quassia wood, Nux-vomica, Gentian Carminatives Coriander, fennel, clove, peppermint Purgatives Senna, Rhubarb Expectorant Tulsi, Balsam of Tolu Cardiotonic Digitalis, Squill, Strophanthus
  • 55. ADVANTAGE  This system of classification can be used for suggesting substitutes of drugs if they are not available at a particular place or point of time. DISADVANTAGE  Drugs having different action on the body gets classified separately in more than one group that causes confusion.  E.g. Cinchona is an antimalarial drug because of presence of quinine but can be put under the group of drug affecting heart because of antiarrythymic action of quinidine.
  • 56.  The crude drugs are divided into different groups according to the chemical nature of their relevant constituent.  The chemical classification of drugs is dependent upon the grouping of drugs with identical constituents.
  • 57. 1. Carbohydrates  Gums - Acacia, Tragacanth, Guargum  Mucilages - Plantago seed  Others include Starch, Honey, Agar, Pectin, Cotton .
  • 58.  Anthraquinone Glycosides - Aloe, Cascara, Rhubarb, Senna  Saponins Glycosides - Quillaia, Arjuna, Glycyrrhiza  Cyanogenetic Glycosides – Bitter almond, wild cherry bark.  Isothiocyanate Glycosides - Mustard  Cardiac Glycosides - Digitalis, Strophantus , squill  Bitter Glycoside- Gentian, quassia, Chirata, Kalmegh.
  • 59.  Tannins are astringent, bitter plant based poly hydroxy phenols that either bind or precipitate proteins. E.g.-pale catechue Black catechue Amla, Behra,Harde, Myrobalan etc.
  • 60.  Monoterpenes and sesquiterpenes obtained from plants.  Examples- Cinnamon, Fennel, Dill, Caraway, Coriander, Cardamom, Orange peel, Mint, Clove
  • 61.  Fixed oils – Castor, Olive, Almond, Shark liver oil  Fats – Theobroma, Lanolin, Lard  Waxes – Beeswax, Spermaceti , Wool fat.
  • 62.  Nitrogenous substances of plant origin  Pyridine and Piperidine – Lobelia, Nicotiana  Tropane - Coca, Belladonna, Datura, Stramonium, Hyoscyamus, Henbane  Quinoline – Cinchona
  • 63.  Isoquinoline – Opium, Ipecac, Calumba  Indole – Ergot, Rauwolfia  Amines – Ephedra  Purine bases – Tea, coffee
  • 64.  Complex mixture of resinol, resin acids, resitannols, resene etc.  Eg. Colophony, podophyllum, jalap, capsicum, turmeric, asafoitida etc.
  • 65. d on an accepted system of botanical classification  Natural relationship  Grouped into: • Kingdom- Plant • Division-Angiosperm • Class- Dicotylendons • Order- Tubiflorae • Family- Solanaceae • Genus- Datura • Species- metal • Variety- fastuosa
  • 66. Plant kingdom Thallophyta Algae, Fungi Eg. Agar Red algae Pteridophyta Derived name from fern Silent feature of plants Rise in pot as ornamentals eg,. Male fern Gymnosperm Gymno- naked Sperm-seed Plant body is sporophytes and differentiate into root, stem and leaves. Eg. Ephedra Angiosperm Enclosed seed Dicot Two cotylendon s Coriander capsicum Monocot One cotylendon s Colchicum Maize
  • 67. Allows for precise and ordered arrangement of drugs.  Accommodates any drug without ambiguity DISADVANTAGES Can not recognize drug is organized or unorganize
  • 68.  Chemo- nature of active constituents of drug  Taxonomy- science deal with identification, nomenclature and classification of plants.  It involves study of chemical variation in different type of plant and used of this information in classification of crude drugs.  Generally secondary metabolites are studied. e.g.. Tropane alkaloids- solanaceae family Isothiocynates- crucifareae Anthocynidines- red, blue and other colors of flower.
  • 69. DRUG PLANT SOURCE USE(S) Vinblastine Catharanthus roseus Anticancer Vinblastine Catharanthus roseus Anticancer Reserpine Rauvolfia serpentina Antipsychoyic, antihypertensive Quinine Cinchona sp. Antimalarial, amoebic dysentery Pilocarpine Pilocarpus jaborandi Antiglucoma Cocaine Lidocaine, procain Erythroxylum coca Topical anesthetic
  • 70. DRUG PLANT SOURCE USE(S) Codeine Morphine Papaver somniferum Antitussive, Narcotic analgesic Atropine Atropa belladonna Spasmolytic Artemisinin Artemesia annua Antimalarial Taxol Taxus baccata, T. brevifolia Breast and ovarian cancer Allicin Allium sativum Antifungal, amoebiasis Podophyllotoxin Etoposide, Teniposide Podophyllum Anticancer
  • 71. Adulteration and Quality control of crude drug c
  • 72. • Inferiority is a natural substandard condition (e.g. where a crop is taken whose natural constituent is below the minimum standard for that particular drug) which can be avoided by more careful selection of the plant material. • Eg- minimum value of strychnine is 1.15% • Less than 1.15% strychnine content - inferior • The term 'adulteration' or debasement of an article covers a number of conditions, which may be deliberate or accidental. Usually in crude drugs, this practice includes substitution of the original crude drugs partially or fully with other substances which is either free from or inferior in therapeutic and chemical properties. • DEFINATION- comes from Latin word which means mixing the original drug material with other false material.
  • 73. • Spoilage is a substandard condition produced by microbial or other pest infestation, which makes a product unfit for consumption, which can be avoided by careful attention to the drying, and storage conditions. • Deterioration Due to age Less chemical component of Drug,also can be due to high temp., light, improper storage condition, bacterial growth or intentional extraction of the constituents and the sale of the residue as the original drugs. • Admixture is the addition of one article to another through accident, ignorance or carelessness e.g. • During collection of root and rhizome, stones are collected. • inclusion of soil on an underground organ or the co- collection of two similar species.
  • 74. • Sophistication intentional addition of other substance in original drug with intent to defraud; such materials are carefully produced and may appear at first sight to be genuine • e.g. powder ginger may be diluted with starch with addition of little coloring material ( turmeric ) to give the correct shade of yellow colour. • Substitution is the addition of an entirely different subastance in replacement of that which is required e.g. supply of cheap cottonseed oil in place of olive oil. • Addition of water in cow milk- adultration • Replacement of goat milk in place of cow milk- substitution
  • 75. • Non availability • Ignorance and inadequate knowledge of collectors • Regional/ linguistic nomenclature • Ecotype, genotype, chemo-type variation • Making profit
  • 76. Different methods used for adulteration may be grouped as follows: 1.Substitution with Inferior Commercial Varieties Due to morphological resemblance to the authentic drugs, different inferior commercial varieties are used as adulterant which may or may not have any chemical or therapeutic potential as that original natural drug. E.g. Arabian Senna (Cassia angustifolia),dog Senna (Cassia obovata) and avaram (Cassia auriculata) have been used to adulterate Senna (Cassia senna); - TYPES OF ADULTERATION OR SUBSTITUTION OF HERBAL DRUGS: TYPES OF ADULTERATION OR SUBSTITUTION OF HERBAL DRUGS: TYPES OF ADULTERATION OR SUBSTITUTION OF HERBAL DRUGS:
  • 77. - Japanese ginger (Zingiber mioga), African and cochin ginger to adulterate medicinal ginger (Zingiber officinale). - Strychnous nux-blanda or S. potatorum in place of S. nux-vomica - Capsicum minimum replaced by C.annuum. - Gentian substituted by kutki 2. Adulteration by Artificially Manufactured Substitutes • To provide the general form and appearance of various drugs, some materials are artificially manufactured and are used as substitute of the original one • Generally this practice is for much costlier drugs.
  • 78. • E.g. - artificial invert sugar for honey; - paraffin wax after yellow coloration substituted for bees wax. - Compressed chicory in place in place ofcoffee - Properly cut and shaped baswood for Nutmeg 3. Substitution by Exhausted Drugs Here the same plant material is mixed which is having no active medicinal components as they have already been extracted out. This practice is most common in case of volatile oil containing materials like clove, fennel, coriander, caraway etc., where the dried exhausted material resembles the same like original drug (similarly with drugs like Cascara sagrada and ginger).
  • 79. Sometimes when coloring matters have been extracted or removed during exhaustion, the residue is re-colored with artificial dyes as is done with saffron and red rose petals. Exhausted ginger made bitter with aloes 4.Substitution by Superficially Similar but Cheaper Natural Substances (Inferior drugs) Usually here the adulterated product has no relation with the genuine article, may or may not have any therapeutic or chemical component desired, Due to their morphological resemblance, they are marketed as adultrants.
  • 80. - leaves of species - Ailanthus are substituted for belladonna, senna, mint etc.; -Leaves of Phytolacca and Scopolia for belladona; -Leaves of Xanthium for stramonium and dandelion for henbane; -Indian dill with European dill or caraway etc. Mother cloves and clovestalks are mixed with clove -Beeswax is substituted by Japan wax. 5. Adulteration by Addition of Worthless Heavy Materials i.e. Harmful adulterants The wastes from market are collected and admixed with authentic drug. This is particularly noticed for liquids or unorganised drugs. E.g. -A large mass of stone mixed with Liquorice root -pieces of limestone are found in asafoetida -
  • 81. - lead shot has occurred in pieces of opium -Pieces of ambered coloured glass in colophony -White oil in coconut oil -Cocoa butter mixed with stearin or paraffin The addition of rodent faecal matter to cardamom seed is a very harmful adulterant. 6.Addition of Synthetic Principles Sometimes to fortify inferior natural products, synthetic principles are added E.g. adding citral to oil of lemon and orange; benzyl benzoate to balsam of Peru etc.
  • 82. 7. Usage of Vegetative Matter from the Same Plant This is done by mixing adventitious matters, miniature plants growing alongwith with medicinal plant in excessive amount or parts of plant other than that which constitutes the drugs due to their resembling colour, odour and in some cases constituents. E.g. The lower plants like moss,liver warts and epiphytes growing in bark portion are mixed with Cascara or Cinchona; - stems of buchu are sometimes cut into short lengths and added to the drug. - Stem portions are mixed alongwith leaf drugs like stramonium, lobelia and senna.
  • 83. 8. Adulteration of Powders Powdered forms are frequently found to be adulterated. E.g. -Dextrin in Ipecacuanha -Powdered liquorice or gentian admixed with powdered olive stones -Exhausted ginger powder in powdered colocynth or ginger -Red sanders wood in capsicum The powdered bark is frequently found to be adulterated with brick powder.
  • 84. EFFECT OF USING ADULTERANTS Effect of using adulteration EFFECT OF USING ADULTERANTS EFFECT OF USING ADULTERANTS Adulterated or spurious raw materials may not possess the therapeutically active constituents, or even may be having some toxic compounds that may not only affect the efficacy of the finished product but also cause deleterious effect on human health. •Reliability of the finished product is at national and international level is affected and betraying the faith of people on the Indian System of Medicine
  • 85. DRUG- EVALUATION Drug Evaluation means conformation of its identity and determination of its quality and purity and detection of nature of adulteration. It is necessary because 1) Biochemical variation in the drug 2) Deterioration due to treatment and storage 3) Substitution and adulteration, as a result of carelessness, ignorance or fraud. Initially, crude drugs were identified by comparison only with the standard description available. Due to advancement, at present evaluation also includes method of estimating active constituent present in the crude drug, in addition to its morphological and microscopic analysis. Its possible to perform physical evaluation which could be both qualitative and quantitative nature
  • 86. Different techniques involved are 1) Morphological and Organoleptic evaluation 2) Microscopic evaluation 3) Chemical evaluation 4) Physical Evaluation 5) Biological Evaluation Standardization (Drug Evaluation) of Crude drugs
  • 87. MORPHOLOGICAL EVALUATION •Organoleptic evaluation means conclusions drawn from studies resulted due to impressions on organs of senses. Done for unorganized drug. • Morphological evaluation means identification of drug by size, shape, specific textute ( done for organized drug) The study of form of crude drug is MORPHOLOGY, while description of the form is MORPHOGRAPHY. E.g. -Fractured surface in cinchona, quillaia, cascara barks and quassia wood
  • 88. (SHAPE) Nux vomica- disc shape Sarpgandha –wavy shape (TASTE)Liquorice & stevia – sweet taste ODOUR- MENTHA & CLOVE- AROMATIC (COLOUR)Cardmom & coriander if shade dried- green colour Sun dried- pale green
  • 89. ORGANOLEPTIC EVALUATION  Aromatic odor of Umbelliferous fruits  -Sweet taste of Liquorice  -Brown color of cinnamon  -Odor and taste of spice-drugs like, asafoetida, black pepper, nutmeg, caraway, cummin etc
  • 90. MICROSCOPIC EVALUATION • Allows more detailed examination of a drug BY USE OF MICROSCOPE • Can be used to identify the organised drugs by their known histological characters (cellular) • Mostly used for qualitative evaluation of organised crude drugs in entire and powdered forms . CHEMOMICROSCOPY: • For the effective results, various reagents or stains can be used to distinguish cellular structure (chemo-microscopy)  A drop of phloroglucinol and conc. HCL give red stain with lignin.  Mucilage is stained pink with Ruthenium red
  • 91. QUANTITATIVE MICROSCOPIC EVALUATIONS- it involves a specific histological characters. LEAF CONSTANT CONTAINS 5 PARAMETERS 1. STOMATAL NUMBER 2. STOMATAL INDEX 3. VEIN ISLET NUMBER 4. VEIN TERMINATION NUMBER 5. PALISADE RATIO •1.LEAF CONSTANT : Stomatal number and index - Why- to identify crude leafy drugs & to distinguish between some closely related species. - Evaluation and purity determination of crude leafy drug - Eg- Indian senna & Alexandrian senna. - i) Stomatal number – is average number of stomata per sq. mm of the leaf. - Example- Indian senna stmatal no. in upper epidermis 220-260, lower epidermis-240-265
  • 92.
  • 93. ii) Vein islet number – is defined as the number of vein- islets per sq. mm of the leaf surface midway between midrib and the margin. Levin in 1929 determined vein islet no. Name of Plant Range Adhatoda vasica 6-8 Datura stramonium 12-16 Digitalis purpurea 2.5-3.0 Eucalyptus globulus 8-13.5 Ocimum sanctum 19-23 2) Stomatal Index – it is the percentage proportion of number of stomata form from the total no. of epidermis cell. Stomatal index are constant. Calculated by using the following equation: S.I. = [S / (E + S) ] X 100 Where, S.I.= Stomatal Index S = No. of stomata per unit area E = No. of epidermal cells in the same unit area
  • 94.
  • 95. iii) Vein-termination number - is defined as the number of veinlet terminations per sq. mm of the leaf surface midway between midrib and margin.
  • 96. Leaf constants or Diagnostic characters of leaves i)Palisade ratio – is defined as average number of palisade cells beneath each epidermal cell. It can be determined with powdered drugs also. Table: Palisade ratios of various leaf drugs Name of Plant Range Adhatoda vasica 5.5-6.5 Datura stramonium 4.2-6.5 Digitalis purpurea 3.7-4.2 Eucalyptus globules 5.5-6.5 (upper) Ocimum sanctum 02-3.5
  • 97.
  • 98. STOMATA It is a minute epidermal opening present on aerial parts of the plants, with followin characteristics: i) A central pore ii) Two kidney shaped similar cells containing chloroplasts known as guard cells and varying number of subsidiary (epidermal) cells covering the guard cells.  Function of stomata is gaseous exchange and the secondary function transpiration.  It is not essential that each plant must have stomata. EG-They are also present in stems (Ephedra), flowers (clove), and fruits (fennel).  It is generally observed that stomata are abundantly present in dicot leaves.
  • 99.
  • 100.  In some cases, they are present on the upper surface of leaves, while in others on lower surface only (coca and cherry).  In some, the stomata are present on both surfaces of the leaves (senna, belladonna, datura etc.).  The distribution of stomata between upper and lower epidermis in dicot leaves shows great variation.  Types of Stomata •Depending upon the guard cells and arrangement of subsidiary cells, stomata are divided into four types: 1. Moss type 2. Gymnospermous type 3. Gramineous type 4. Dicotyledonous type
  • 101. ACCORDING TO ARRANGEMENT OF SUBSIDARY CELLS: i) Paracytic or Rubiaceous stomata This type of stomata comprises of two guard cells covered by two subsidiary cells, the long axis of which are parallel to that of stoma e.g. coca and senna leaves ii) Diacytic or caryophyllaceous or cross-celled stomata The guard cells are covered by two subsidiary cells, as in case of paracytic stoma, but the arrangement of subsidiary cells on the guard cell is at right angle to that of stoma, e.g. Peppermint, spearmint and Vasaka
  • 102.
  • 103. iii) Anisocytic or cruciferous or unequal celled stomata: The guard cells are covered by three subsidiary cells, of which one is markedly smaller than the other two, e.g. Belladonna, datura and stramonium iv) Anomocytic or ranunculaceous or irregular celled stomata: Stoma is surrounded by varying number of subsidiary cells resembling other epidermal cells e.g. Digitalis, lobelia, buchu IP recognise one more type of stomata, v) Actinocytic or radiate celled stomata Two guard cells are surrounded by a circle of radiating subsidiary cells.
  • 104. TRICHOMES • Trichomes are the tubular elongated or glandular outgrowth of the epidermal cells . • Trichomes are also called as plant hairs. trichomes consists of two parts root and body. • Ttrichomes present in most of plant parts and are function less but some times perform secretory function. • Depending up on the structure and the number of cells present in trichomes,they are classified in to following. 1.Covering Trichomes or non glandular trichomes: 2.Glandular Trichomes: eg dhatura 3.Hydathode or special Trichomes: Trichomes absent in coca, less in senna leaves.
  • 105.
  • 106.
  • 107.
  • 108.
  • 109.
  • 110.
  • 111.
  • 112.
  • 114. QUANTITATIVE MICROSCOPY Lycopodium spore method: It is use to determination of percentage of foreign organic matter, mainly use for powdered drugs. Developed by T.E WALLIS. • It is used when especially chemical and other methods of evaluation of drugs fails to determine quality. • Lycopodium spores are very characterized in shape and appearance and uniform in size (25μm) on avg., 94000 spores present/mg of lycopodium powder. • It consists of 1.well defined particles which may be counted. 2.Single layered cells or tissues the area of which may be traced under suitable magnification and actual area calculated 3.The objects of uniform thickness, the length of which can be measured, and actual area calculated.
  • 115.
  • 116.
  • 117. PHYSICAL EVALUATION These are rarely constant for crude drug , but may help in evaluation with reference to crude drug. 1. Moisture content 2. Specific gravity 3. Density 4. Optic rotation 5. Refractive index 6. Melting point 7. Viscosity 1. Moisture content is responsible for the decomposition of crude drug due to chemical change or microbial attack. It is necessary to determine and control the moisture content of crude drug . It is determined by heating the drug at 105°C in an oven to a constant weight. Moisture content of is 5% w/w.
  • 118. 2. Viscosity: Resistance of the fluid to flow . Viscosity of a liquid is constant at given temperature . Hence it is used as a means of standardizing liquid drugs. PYROXYLIN kinematic viscosity is 1100-2450 centistokes . LIQUID PARAFFIN kinematic viscosity is 64 centistokes at 37.8°C. 3. Melting Point: It is one of the parameter to judge the purity of crude drug. In case of pure chemical or phytochemicals , melting point are very sharp and constant . Since the crude drug from animals and plants origin contain the mixed chemicals . Purity of crude drug can be determined by their melting points. Eg. COLOPHONY 75-85°C, BEES WAX 62-65°C, WOOL FAT 34-44°C COCOA BUTTER 30-33°C. Certain drugs fluoresce when the cut surface or the powder is exposed to
  • 119. Certain drugs fluoresce when the cut surface or the powder is exposed to ultraviolet radiation , And it is useful in the identification of those drugs . Indian and Chinese Rhubarb are very difficult to distinguish and is very difficult form , but examination in ultraviolet light gives such marked differences in florescence that the varieties can be easily distinguish from each other . 4. Solubility: The number of ml of solvent required to dissolve 1g of drug . The presence of adulteration in a drug could be indicated by solubility studies. Eg. Balsam of peru is soluble in chloral hydrate solution, Colophony is freely soluble in light petroleum, Asafoetida is soluble in carbon disulphide., Alkaloidal bases are soluble in chloroform.
  • 120. The following are the most important parameter with respect to crude drug. Ash Value: The residue remaining after incineration is the ash content of the drug (inorganic salts of carbonates , phosphates , silicates of sodium , potassium, calcium and magnesium ) is known as ash content. Ash value is the criterion to judge the identity and purity of crude drug. It is also useful for detecting low grade products, exhausted drug and drugs from earthy matter . There are three types of Ash value: Total Ash Acid Insoluble Ash Water soluble Ash
  • 121. The total ash includes both “physiological ash”, which is derived from the plant tissue itself, and “non-physiological” ash, which is the residue of the extraneous matter (e.g. sand and soil) adhering to the plant surface. Acid-insoluble ash is the residue obtained after boiling the total ash with dilute hydrochloric acid, and igniting the remaining insoluble matter. This measures the amount of silica present, especially as sand and siliceous earth. Water-soluble ash is the difference in weight between the total ash and the residue after treatment of the total ash with water.
  • 122. Extractive Value • It is used to detect either material exhausted by water or not (tea leaves , ginger rhizomes) The extract obtained by exhausting crude drugs with different solvents are approximate measures of their chemical constituents . • Various solvents are used according to the type of the constituents to be analyzed . • Useful for the evaluation especially when the constituents of the drugs can not be readily estimated by any other means . • It also helps to indicate the nature of chemical constituents present in the drug. • Also helps in the identification of adulteration of drug . Solubility of drug is also detected . • There are three types of extractive value. Water soluble extractive value Alcohol soluble extractive value Ether soluble extractive value.
  • 123. 1. Water soluble Extractive It is applied for the drugs which contain water soluble constituents such as tannins , sugar , plant acids and mucilage. 2. Alcohol soluble Extarctive It is applied for drugs which contain alcohol soluble constituents such as tannins, resins and alkaloids. 3. Ether soluble Extractives: It is applied for the extraction of volatile oils , fixed oils and resins . Volatile ether soluble extractive value Non volatile ether soluble extractive value. .
  • 124. Foreign Organic matter : The parts of the organ or organs other than those named in the definition and description of the drug are defined as foreign organic matter. The maximum limit for the foreign organic matter is defined in the monograph of crude drug. If it exceedsthe limits, deterioration in quality of the drug takes place. Swelling factor: Significances : Useful in the evaluation of crude drugs containing mucilage Useful for the detection of purity of the crude drug Determination : 1. Transfer 1 gm of the seeds to a 25ml stoppered cylinder 2. Fill up to the 20ml mark on the cylinder with water. Agitate gently and occasionally during 24 hours and allowed to stand 3.Measure the volume occupied by the swollen seeds. and occasionally during 24 hours and allowed to stand 3.Measure the volume occupied by the swollen seeds.
  • 125. Spectrophotometric methods: i) UV- Ultra violet /visible spectroscopy ii)IR-Infra Red spectroscopy iii) Fluorescence analysis iv) NMR-nuclear magnetic resonance spectroscopy v) MS-Mass spectroscopy vi) X-ray diffraction vii) RIA-radio immuno assay. Above mentioned all spectrphotometric evaluation techniques are useful for structure elucidation of Phytoconstituents.
  • 126. Chromatographic techniques: TLC-Thin layer chromatography Principle :Adsorption Adsorbent silica gel G/C coated to a thickness of minutes and used. After development of chromatography spots are revealed by spraying with suitable detecting agent TLC is useful to analyse Alkaloids, Glycosides like all bio- constituents. The Rf value vary depend on the nature of substance, composition of solvent and impurities. Rf value = TLC/HPTLC are micro analytical techniques used for determination of natural products Advantages : simple in operation and rapid Chromatographic techniques.
  • 127. CHEMICAL EVALUATION Determination of the active constituent in a drug by chemical tests is referred to as chemical evaluation. The following are various methods of chemical evaluation 1. Instrumental methods 2. Chemical tests 3. Individual constituent chemical tests 4. Micro chemical tests 1. Instrumental methods: They make use of various instruments for evaluation like colorimetry, flourimetry, spectrophotometry etc. 2. Chemical constants tests: These are like acid value, iodine value and ester value etc are used for the identification of fixed oils and fats. 3. Individual chemical tests: These are the tests which are used for identifying particular drugs. 4. Micro-chemical tests: These are the tests which are carried on slides. Example: Euginol in clove oil is precipitated as potassium euginate crystals.
  • 128. Method for chemical evaluation Extract obtained using petroleum ether, chloroform, ethanol and water was prepared using the respective solvent. These extracts along with positive and negative controls were tested for the presence of active phytochemicals viz: tannins, alkaloids, phytosterols, triterpenoids, falvonoids, cardiac glycosides, anthroquinone glycosides, saponins, carbohydrates, proteins, amino acids and fixed oils & fats following standard methods Tannins 1. Ferric chloride Test: Added a few drops of 5% ferric chloride solution to 2 ml of the test solution. Formation of blue color indicated the presence of hydrolysable tannins. 2. Gelatin Test: Added five drops of 1% gelatin containing 10% sodium chloride to 1 ml of the test solution. Formation of white precipitates confirmed the test.
  • 129. Alkaloids Approximately 50 mg of extract was dissolved in 5 ml of distilled water. Further 2M hydrochloric acid was added until an acid reaction occurred and filtered. The filtrate was tested for the presence of alkaloids as detailed below 1.Dragendorff’s Test: To 2 ml of the filtrate was added 1 ml of Dragendorff’s reagent. Formation of orange or reddish brown precipitate indicated the test as positive. 2.Mayer’s Test: To 1 ml of test solution or filtrate was added a drop or two of the Mayer’s reagent. white or a creamy precipitate confirmed the test as positive. 3.Hager’s Test: To 1 ml of test solution or filtrate, a drop or two of Hager’s reagent formation of yellow precipitate indicated the test as positive. 4.Wagner Test: Two drops of Wagner’s reagent was added to 1ml of the test solution. The formation of yellow or brown precipitate confirmed the test as positive for alkaloids.
  • 130. Phytosterols 1. Liebermann-Burchard’s Test: The extract (2 mg) was dissolved in 2 ml of acetic anhydride, heated to boiling, cooled and then 1 ml of concentrated sulfuric acid was added. A brown ring formation at the junction and the turning of the upper layer to dark green color confirmed the test for the presence of phytosterols. Triterpenoids Salkowski Test: Approximately 2 mg of dry extract was shaken with 1 ml of chloroform and a few drops of concentrated sulfuric acid were added.A red brown color formed at the interface indicated the test as positive for triterpenoids. Saponins 1.Foam Test: 5 ml of the test solution taken in a test tube was shaken well for five minutes. Formation of stable foam confirmed the test. 2. Olive oil test: - Added a few drops of olive oil to 2ml of the test solution and shaken well. The formation of a soluble emulsion confirmed the test.
  • 131. Flavonoids 1.Shinoda test: A few magnesium turnings and 5 drops of concentrated hydrochloric acid was added drop wise to 1 ml of test solution. A pink, scarlet, crimson red or occasionally green to blue color appeared after few minutes confirmed the test. 2. Alkaline reagent test: Addition of 5 drops of 5% sodium hydroxide to 1 ml of the test solution resulted an increase in the intensity of the yellow color which became colorless on addition of a few drops of 2 M hydrochloric acid which indicated the presence of falvonoids. 3.Lead acetate test: A few drops of 10% lead acetate added to 1ml of the test solution resulted in the formation of yellow precipitate confirmed the presence of falvonoids.
  • 132. Cardiac glycosides 1.Keller -Killiani test: Added 0.4 ml of glacial acetic acid and a few drops of 5% ferric chloride solution to a little of dry extract. Further 0.5 ml of concentrated sulfuric acid was added .The formation of blue color in acetic acid layer confirmed the test.. Carbohydrates 1.Molisch’s test: To 1 ml of test solution added a few drops of 1 % alpha-napthol and 2-3 ml concentrated sulfuric acid. The reddish violet or purple ring formed at the junction of two liquids confirmed the test. 2. Barfoed’s test: 2ml of reagent was added to 2 ml of the test solution, mixed & kept a in boiling water bath for 1 min. Red precipitate formed indicates the presence of monosaccharide's.
  • 133. 3. Seliwanoffs test: To 3 ml of Seliwanoffs reagent was added to 1 ml of the test sample and heated on a water bath for one minute. The formation of rose red color confirmed carbohydrates 4. Fehlings test: Dissolved 2 mg dry extract in 1 ml of distilled water and added 1ml of Fehling’s(A+B) solution, shooked and heated on a water bath for 10 minutes. The brick red precipitate formed confirmed the test. Anthraquinone glycosides Hydroxyanthraquinone Test: To 1 ml of the extract, added a few drops of 10% potassium hydroxide solution. The Formation of red color confirmed the test. .
  • 134. Test for proteins: Biuret test: To 2 ml of the test solution added 5 drops of 1% copper sulphate solution and 2 ml of 10% NaOH .Mix thoroughly. Formation of purple or violet color confirmed proteins. Test for amino acids 1. Millon’s test: Added 5 drops of millons reagent to 1 ml of test solution and heated on a water bath for 10 min, cooled and added 1% sodium nitrite solution. Appearance of red color confirmed the test. Fats and fixed oils : To 5 drops of the sample was added 1 ml of 1% copper sulphate solution and a few drops of 10% sodium hydroxide. The formation of a clear blue solution confirmed the test.
  • 135. BIOLOGICAL EVALUATION • It is employed when the drug cannot be evaluated satisfactorily by chemical and physical methods. • In this method, the response produced by the test drug on a living system is compared with that of the stranded preparation. • Such an activity is represented in units as International Units (I.U). Dose is termed as International units IU eg. Digitalis 1IU=76mg of standard ,Vit-A 1IU=0.344 of standard , Vit-D 1IU=0.025of standard.
  • 136. Indication of Biological Evaluation • When the chemical nature of the drug is not known but is has an biological action. • When chemical methods are not available. • When the quantity of the drug is small and so it cannot be evaluated chemically. • Drugs which have different chemical composition but same biological activity. Example: Cardiac glycosides are evaluated by this method on cats, frogs or pigeons.
  • 137. SIGNIFICANCE 1.The method is generally used when standardization is not done satisfactory by chemical or physical methods 2.When the quantity of the drug /sample are very less then the drugs are evaluated by biological methods. 3.These methods are performed on living animals, isolating living organ and tissue, animal preparation, and micro-organism ( Bioassay). METHODS OF STUDIES 1)Toxic----animals are used 2)Symptomatic-----animals are used 3)Tissue-------isolated tissue is used To estimate potency of drug ,With entire animal or with tissue, To conform therapeutic activity.
  • 138. Evaluation of Hypoglycemic activity deficiency of glucose in the bloodstream. • Traditional Diabetic drugs -Momordica charantaka,Fenu greek,Gudmar. Diabetis is induced in animals by Alloxan & Streptazocin. • Alloxan cause necrosis of pancreatic islet-B cells which shows 180-250mg/ml fasting blood glucose levels • Streptazocin cause formation of streptomycin they produce cytotoxic nitrourcido glucopyranose which cause diabetes • ANIMALS USED: Rabbits,Rats,Mice—(4 to 7 days) Dose :rats--80mg/kg ,mice— 150mg/kg of streptazocin single oral injection 140-180 mg/kg of alloxan for rabbits at marginal ear vein for 7 days for rats and mice intraperitoneally 2 days Insulin levels are noted by tests like RIA.ELISA. •RIA-detects antibody antigen reaction by using radioisotopes. •ELISA-detects antibody antigen reaction by using enzyme.
  • 139. Reference WHO guidelines for assessing quality of herbal medicines By World Health Organization THANK YOU