B.pharm. Class Notes

B.PHARM.
CLASS NOTES
AND
HANDOUTS
• Conservation is the process of management of
biosphere in order to obtain the greatest benefit for
the present generation and maintaining the potential
for future.
• Conservation of medicinal plant resources is of global
concern because we don't know what we are losing
and what we will need in future.
DRUG METABOLISM
Presented by
Saroj kanta Bisoyi
Asst.Professor
RCPHS, BAM
INTRODUCTION
 Biotransformation:
Chemical alteration of the drug in body that converts nonpolar or
lipid soluble compounds to polar or lipid insoluble compounds.
 Drug metabolism may be defined as the biochemical modification

of one chemical form to another, occurring usually through
specialised enzymatic systems.
 It often involves the conversion of lipophilic chemical

compounds (drugs) into highly polar derivatives that can be easily
excreted from the body.
FUNCTION OF BIOTRANSFORMATION
1) It causes conversion of an active drug to inactive or less active metabolite(s)
called as pharmacological inactivation.
Example of drug follow by this mechanism.
Phenobarbitone P-hhdroxy phenobarbitone
Phenytoin P-Hydroxy phenytoin
Procaine P-amino benzoic acid
2) It causes conversion of an active to more active metabolite(s) called as
bioactivation or toxicological activation.
Codeine Morphine
Halothane Trifluoro acetic acid
paracetamol N –Hydroxylation derivative
3) It causes conversion of an Inactive drug active metabolite.

Ex of drug follow by this mechanism.
Levodopa- dopamine
SITE/ORGANS OF DRUG METABOLISM
 The major site of drug metabolism is the liver (microsomal

enzyme systems of hepatocytes).
 Secondary organs of biotransformation
➢ kidney (proximal tubule)
✓ lungs
➢ testes
➢ skin
➢ intestines
LIVER
 The primary site for metabolism of almost all drugs
because it is relatively rich in a large variety of
metabolising enzymes.
 Metabolism by organs other than liver (called as extra-

hepatic metabolism) is of lesser importance because lower
level of metabolising enzymes is present in such tissues.
 A few drugs are also metabolised by non-enzymatic means

called as nonenzymatic metabolism.
 The drug metabolising enzymes can be broadly divided

into two groups: microsomal and non-microsomal
enzymes.
TYPE OF DRUG METABOLISM
 The metabolism of drug in the body is achieved by two types
of reactions .these reactions are
1) Phase-I reaction
2) Phase-II reaction
Phase 1 reaction. (Non synthetic phase).
➢ a change in drug molecule. generally results in the

introduction of a functional group into molecules or the
exposure of new functional groups of molecules.
PHASE –I REACTION
❖ In this reaction a functional polar group
(ie-OH,COOH,NH2,SH) is introdused to drug or in
xenobiotics to convert them into more water
soluble compounds so they are easily excreted out
from the body.
Phase-1 reaction convert the parent drug into polar
metabolites by three ways.
a) Oxidation reaction
b) Reduction reaction
C) Hydrolysis reaction
A) OXIDATION REACTIONS
 Addition of oxygen/ negatively charged radical or
removal of hydrogen/ positvely charged radical.
 Reactions are carried out by group of enzyme
monooxygenases in the liver.
 Fianl step: Involves cytochrome P-450 haemoprotein,
NADPH, cytochrome P-450 reductase and O2

CYTOCHROME P FAMILY
 Multiple CYP gene families have been

identified in humans, and the categoriezed
based on protein sequence homology
 Most of the drug metabolizing enzymes are

in CYP 1, 2, & 3 families .
 CYP3A4 is very common to the metabolism

of many drugs.
CYTOCHROMES: METABOLISM OF DRUGS
CYP Enzyme Examples of substrates
1A1 Caffeine, Testosterone, R-Warfarin

1A2 Acetaminophen, Caffeine, Phenacetin, R-Warfarin
2A6 Estradiol, Testosterone
2B6 Cyclophosphamide, Erythromycin, Testosterone
2C-family Acetaminophen, Tolbutamide (2C9); Hexobarbital,
SWarfarin (2C9,19); Phenytoin, Testosterone, R-
Warfarin, Zidovudine (2C8,9,19);
2E1 Acetaminophen, Caffeine, Chlorzoxazone, Halothane
2D6 Acetaminophen, Codeine, Debrisoquine
3A4 Acetaminophen, Caffeine, Carbamazepine, Codeine,
Cortisol, Erythromycin, Cyclophosphamide, S- and R
Warfarin, Phenytoin, Testosterone, Halothane
NON-CYP DRUG OXIDATIONS
 Monoamine Oxidase (MAO), Diamine Oxidase (DAO)
➢ MAO (mitochondrial) oxidatively deaminates endogenous

substrates including neurotransmitters’
➢ Dopamine, serotonin, norepinephrine, epinephrine
 Alcohol & Aldehyde Dehydrogenase
➢ Non-specific enzymes found in soluble fraction of liver
➢ Ethanol metabolism
TYPES OF DRUG METABOLISM OXIDATION
REACTIONS
 Aromatic and side chain hydroxylation.
 Oxidation of benzylic carbon atom
 Oxidation of allylic carbon atom
 Oxidation at the carbon alpha to carbonyl and imino group.
 Oxidatiive N and O-dealkylation
 N- oxidation and sulphur oxidation.
 Desulphuration
1) AROMATIC AND SIDE CHAIN HYDROXYLATION.
 Hydroxylation means addition of –OH group to the

aromatic ring.
 The reaction involves the formation of an epoxides as

an intermediate and under goes rearrangement to
form phenolic products
1) AROMATIC AND SIDE CHAIN HYDROXYLATION.
5) OXIDATIVE N AND O-DEALKYLATION
 oxidative N and O-dealkylation involves hydroxylation of
alpha carbon adjecent to amino group and oxygen to form
carbinolamine or hemiactal as an intermedite than
undergoes clevage to release the alkyl group and the drug
metabolites.
5) OXIDATIVE N AND O-DEALKYLATION
REDUCTION
 Nitro,azo and carbonyl group containg drug are easily

redused by various enzyme present in the body
(reductase enzyme).
reduction of aromatic nitro group reduced to primary
amine as metabolites.
AZO GROUP CONTAINING DRUGS UNDERGOES
ENZYMATIC REDUCTION GIVE TWO AMINES.
CARBONYL CONTINING DRUG ON ENZYMATIC
REDUCTION GIVES ALCOHOL AS METABILITES.
HYDROLYSIS REACTION
 Drugs containing ester or amide functional group

undergoes metabolism by this pathways.
HYDROLYSIS
 Drugs containing ester or amide functional group

undergoes metabolism by this pathways.
AMIDE DRUGS LIKE LIGNOCAINE
UNDERGOES HYDROLYSIS TO FORM AMINE
 Hydrolysis of amide is very slow as compared to

esters.
 The drug produced by hydrolysis get easily
excreted as compared to parent drugs.
 These products have high polarity and easily
show conjugation reactions.
PHASE-II DRUG METABOLISAM
 These are also called conjugation reaction.
 Last step in detoxification reactions and almost always results

in loss of biological activity of a compound.
 May be preceded by one or more of phase I reaction .
 The products formed after phase-II reactions are generally

inactive.
 It involve conjugation with various endogeneous substances

like glucuronic acid,amino acids,glutathion etc.
PHASE –II METABOLISM OCCURS BY
FOLLOWING PATHWAYS
 1) Glucuronidation
 2) Glutathione conjugation
 3) Sulphate conjugation
 4) Methylation
 5) Acetylation
 6) Aminoacid conjugation
1) GLUCURONIDATION
 Also known as glucuronic acid conjugation.
 Phase-1 metabolites having free alcoholic or phenolic groups

cojugates with glucuronic acid to form ether or ester
glucuronides.
 This process is done by the help of the enzyme glucuronyl

transferase.
1) GLUCURONIDATION
 The transfer of glucuronyl group to the drug takes place by the
coenzyme UDPGA (uridine diphospho-α-d-glucuronic acid).
 Glucuronides formed are normally non toxic,highly polar and

easily excreted in the urine or biles.
 It mainly occurs in many tissues like kidney,skin,intenstine,lungs

and brain.
4-HYDROXY PHENYTOIN FORM IN PHASE-1
REACTION FORM PHENYTOIN –O-
GLUCURONIDE
MORPHINE PRODUCES MORPHINE GLUCURONIDES.
GLUTATHIONE CONJUGATION
 Also known as mercaptouric acid is a thiol containing

tripeptide.γ-glutamyl cysteine glycine
 Electron deficient metabolites like alkyl aryl

halides,sulphates,sulphonates,nitrates produced by phase-I
metabolism conjugates with glutathione.
 That catalysed by the enzyme glutathione transferase.

METABOLISM OF ETHACRYNIC ACIDS
SULPHATE CONJUGATION
 Drug having hydroxy groups,phenols and aromatic amines

under goes sulphate conjugation.
 It occurs by the enzyme sulphotransferase and coenzyme

PAPS(Phosphoadenosine phosphosulphate).
 Mostly the steroidal drugs undergoes by this reaction
 The proces occurs mostly in the liver ,kidney and intenstine.
 Ex-Estrone on metabolism produces estrone sulphate.

ESTRONE METABOLISM
METHYLATION
 Most of the endogeneous amine are metabolise by this
methylation conjugation.
 Mostly methyl group is attached with the help of coenzyme
SAM(S-adenosyl methionine) and enzyme methyl transferase.
ACETYLATION
 Acetylation inlvoves conjugation reaction with
acetyl coA by using the enzyme N-Acetyl
transferase present in liver ,lngs,spleen and
gastric cell and red blood cells.
 Drug contains Hydrazino functional group
undergoes acetylation.
 Ex- Isoniazid gets acetylted to form acetyl isoniazid
AMINO ACIDS CONJUGATION
 Drugs having aromatic acids and aryl alkyl acids as
functional group undergoes conjugation with amino acids like
glycine and glutamic acids.
 Metabolites of phase-I reaction having carboxyl group also
conjugates with glycine.
 Ex-Benzoic acid conjugates with glycine to form hippuric acid
DIPHENHYDRAMINE METABOLISM
FACTROR AFFECTING METABOLISM
 Some physical,chemical and biological factors affects the

metabolism of a drugs. These are
 1) Physiochemical properties of the drug molecules.
✓ physiochemical properties like size,,shape,acidity and

basicity,lipophilicity,solubility,pka value of drugs molecules
affects drug metabolism.
2) CHEMICAL FACTORS
 Various chemical affects the metabolism of the drugs.
these are
a) Enzyme inducer
b) Enzyme inhibitors
A) ENZYME INDUCER
 These are the chemical which increases metabolism.
 Ex-3-methyl cholanthrene and cigarette smoke increases
the metabolism of some drugs.
 Alcohol increases the metabolism of coumarins and
phenytoin.
 Barbiturates increases metabolism of oral contraceptives

ENZYME INHIBITORS
 The chemicals and drugs which decreases the

activity of metabolism.
 MAO inhibitors decreases metabolism of barbiturates .
 coumarins dccreases the biotransformation of phenytoin.
 Various halogenated pesticides like DDT,organophasphate

insecticides,heavy metals like mercury,tin ,nickel,cobalt
and arsenic decreases the metabolism of various drugs.
3.ENVIROMENTAL FACTORS
 Enviromental factors like pressure,temperatue and humudity
also affects the metabolism.
4. Biological factors.
Various biological factors which affect metabolism of
drug are
a) Age of the patient
b) Sex of the patient
c) Diet
d) Altered physiological state like pregnancy,diseases

state and hormonal imbalance.
STERIOCHEMICAL ASPECTS OF THE DRUG
MOLECULES.
 Steriochemistry of the drug also affcets the

metabolism.
 Metabolising enzymes have different preference
for one enantiomer than the other.
 Ex-
1. (–) Quinine treat the malaria fever but (+)

quinine does not.
2. D (+) get easily metabolised in the body to give
CO2 and water but L (-) glucose is not metabolised and
excreted as such.
REFERENCES
 Goodman and Gilman, Pharmacological basis of
Therapeutics, 12th edition, Laurence L Bruton
 Essential of Medical Pharmacology, K D Tripathi, 5th
Edition, JP publishers, New Delhi.
 Wilson and Gisvold’s Textbook of Organic Medicinal
and Pharmaceutical Chemistry 11th ed. Lippincott,
Williams & Wilkins ed
 Drug metabolism by S.P. Markey, NIH, accessed on
internet on 02-03-2013 from
www.cc.nih.gov/.../ppt/drug_metabolism_20062007.ppt
 Drug metabolism and pharmacokinetics in drug
discovery: a primer for bioanalytic study: chandrani
gunaratna, current separations, 19:1, 2000
• Edible Vaccine involves, introduction of selected desired
genes into plant and then inducing these altered plants
to manufacture the altered protein.
Antigen in transgenic plant
Ingestion
Delivered by bio-encapsulation
Taken up by Microbial cell
Pass on to the Macrophage
IgG,IgE responses
Local IgA response & Memory cells
Neutralize the attack by the real infectious agent
ADVANTAGE AND DISADVANTAGE OF DIFFERENT PLANTS
Potato
Advantage Disadvantages
• Easily transformed. • Spoils readily.
• Easily propagated. • Need cooking
• Stored for long periods without which denature
refrigeration. antigen
• Grow quickly.
• Cultivate broadly.
• High content Vitamin-A may boost
immune response.
BANANA
• Do not need cooking. • Trees take 2-3 to
• Protein not destroyed even after mature years.
cooking. • Spoils rapidly after
• Inexpensive . ripening.
• Grown widely in developing countries.
RICE
• Commonly • Grows slowly.
used in baby • Requires glasshouse condition.
food. • Do not need cooking.
• High • Protein not destroyed even after cooking.
expression of • Inexpensive .
antigen. • Grown widely in developing countries.
Clinical Trial on
• First human trials of potato-based vaccine against
Hepatitis B have reported encouraging results.
Hepatitis • The amount of HBsAg needed for one dose could
-B be achieved in a single potato.
• Levels of specific antibodies significantly exceeded
the protective level of 10mIU/mL in human.
• 11 Volunteers were feed raw transgenic Potatoes

ETEC expressing LT-B.
• 10 (95%) of these individuals developed neutralizing
antibodies and 6 (55%)develop mucosal response.
• 20 people fed with transgenic potato.

Norwalk
Virus • 19 (95%)of them expressing Norwalk
virus antigen showed seroconversion.
ADVANTAGES OF DISADVANTAGES OF
EDIBLE VACCINES EDIBLE VACCINES
• Cost effective. • Transgenic
• Easy to administer. contamination can
• Easy to store. occur.
• Acceptable to poor developing • Antibiotic resistance
country.
• Fail safe marker genes can
• Activate both mucosal and spread from GM food to
systemic immunity. pathogenic Bacteria.
• Heat stable. • Difficulty in dose
• Do not required cold chain maintenance.
maintenance.
• No fear of contamination.
FUTURE OF EDIBLE VACCINE CONCLUSION: Edible plant
• Resistance to GM foods may derived vaccine may lead to
affect future of Edible Vaccine. a future of safer and more
effective immunization.
• Edible Vaccine involves, introduction of selected desired
genes into plant and then inducing these altered plants
to manufacture the altered protein.
Antigen in transgenic plant
Ingestion
Delivered by bio-encapsulation
Taken up by Microbial cell
Pass on to the Macrophage
IgG,IgE responses
Local IgA response & Memory cells
Neutralize the attack by the real infectious agent
Potato
• Easily transformed. • Spoils readily.
• Easily propagated. • Need cooking
• Stored for long periods without which denature
refrigeration. antigen
• Grow quickly.
• Cultivate broadly.
• High content Vitamin-A may boost
immune response.
BANANA
• Do not need cooking. • Trees take 2-3 to
• Protein not destroyed even after mature years.
cooking. • Spoils rapidly after
• Inexpensive . ripening.
• Grown widely in developing countries.
RICE
• Commonly • Grows slowly.
used in baby • Requires glasshouse condition.
food. • Do not need cooking.
• High • Protein not destroyed even after cooking.
expression of • Inexpensive .
antigen. • Grown widely in developing countries.
Clinical Trial on
• First human trials of potato-based vaccine against
Hepatitis B have reported encouraging results.
Hepatitis • The amount of HBsAg needed for one dose could
-B be achieved in a single potato.
• Levels of specific antibodies significantly exceeded
the protective level of 10mIU/mL in human.
• 11 Volunteers were feed raw transgenic Potatoes

ETEC expressing LT-B.
• 10 (95%) of these individuals developed neutralizing
antibodies and 6 (55%)develop mucosal response.
• 20 people fed with transgenic potato.

Norwalk
Virus • 19 (95%)of them expressing Norwalk
virus antigen showed seroconversion.
ADVANTAGES OF DISADVANTAGES OF
EDIBLE VACCINES EDIBLE VACCINES
• Cost effective. • Transgenic
• Easy to administer. contamination can
• Easy to store. occur.
• Acceptable to poor developing • Antibiotic resistance
country.
• Fail safe marker genes can
• Activate both mucosal and spread from GM food to
systemic immunity. pathogenic Bacteria.
• Heat stable. • Difficulty in dose
• Do not required cold chain maintenance.
maintenance.
• No fear of contamination.
FUTURE OF EDIBLE VACCINE CONCLUSION: Edible plant
• Resistance to GM foods may derived vaccine may lead to
affect future of Edible Vaccine. a future of safer and more
effective immunization.
FACTORS INVOLVED IN THE
PRODUCTION OF CRUDE DRUGS
I. P. Padhy
• Time of collection
• Season of collection
• Ontogenic (age) factor
• Methd of of collection
• Other conditions
Altitude: Height from sea level
• Altitude is a very important factor in cultivation and crude
drug production.
• Tea, Cinchona, eucalyptus requires high altitude.
• In case of Cinchona succirubra the plant grow well at low
levels but practically produce no alkaloids.
Temperature:
• Temperature is a major factor controlling the development
and metabolism of plants. Although each species has
become adapted to its own natural environment, plants
frequently able to exist in a considerable range of
temperature.
• Variation in night and day temperature must also be
considered
• The variation often being considerable, the extent to which
such variation can influence growth
• Hyocyamus muticus.
Rain fall:
• Except the xerophytes like aloe, acacia, most
of the plants need sufficient rain fall or
proper irrigation for their favorable
development.
• The important effects of rainfall on
vegetation need to be considered in relation
to the annual rainfall, its distribution through
out the year.
• Continuous rain can lead to a loss of
aqueous soluble substances from leaves
and roots by leaching.
Day length and radiation characteristics:
• Plants growth vary much in both the amount and intensity
of the light which they require.
• In wild state the plant will be found where its shade
requirements are met, and under cultivation similar shade
must be provide.
• It is observed that light is a factor which helps to
determine the amount of glycosides or alkaloids produced
with Belladonna, Stramonium and Cinchona ledgeriana.
• Full sun shine gives a higher content of alkaloids than
shade.
• Type of radiation has been studied in respect to
morphological development of the plants. Many plants
initiate flowers only in certain day lengths, and where
flowering is essential this factor must be care fully
considered before panting in a new region.
Soil and its fertility:
Different plant species vary enormously in their soil
and nutritive requirement, and this aspect has
received considerable attention with medicinal
plants.
Three basic properties of soils are:
i. Physical
ii. Chemical
iii. Microbiological.
A) Physical Properties:
Depending upon the Depending upon the
size of the mineral matter percentage covered by clay,
the following names are given soil is classified as given
to soil (It is one factor which below.
influence the water holding capacity
and mechanical strength)
Particle size Types of soil Types of Percentage
Soil
Less than 0.002 Fine clay Clay More than 50.0% of
mm clay
0.002 to 0.02 mm Course clay or Loamy 30 to 50% of clay
slit
0.2 to 2.00 mm Course sand Slit loam 20 to 30% of clay
Sandy soil More than 70%

sand
Calcareous More than 20% of
• A soil is good for plant growth should have half of the pore
spaces filled with water and the rest with air, since good
aeration is essential for root development.
• The commonly known soil is shallow upper layer and is the
friable material in which plants find foot hold and
nourishment.
• Clay is one of the highly weathered portions of the soil,
consisting of finest particles. This provides to the soil
adhesive and cohesive properties and also holds plant
nutrients.
B) Chemical properties:
• Any type of soil containing not less than 0.5% of organic
matter is described as rich.
• The highest availability of plant nutrients is in between the
PH range of 6.5 to 7.5.
• To bring the PH to the normal, acidic soils can be limed or
alkaline soils can be reclined by application of gypsum.
• Soil containing more humus and little lime are inclined to
become acid and those with abundant lime are alkaline.
• All plants require calcium as their nutrition but plants known
as calciphobous cannot be grown on chalky soil.
• In some cases different varieties of the same species may
grow on different soils.
C) Microbiological properties:
Little work appears to be performed on the microbiology of soil with respect to
secondary metabolism.
 Soil fertility is the capacity of the soil to provide
nutrients in adequate amount and in balanced
proportion to plants.
 Soil fertility can be maintained by addition of animal
manure, nitrogen fixing bacteria or by application of
chemical fertilizers.
 For vegetative growth, plants need Sunlight, Co2, water,
mineral matter.
 Plants need 16 nutrient elements for synthesizing
various compounds.
• Primary nutrients: Nitrogen, Phosphorous, and
Potassium.
• Secondary Nutrients: Magnesium, calcium and
sulphur.
• Trace elements: Copper, Manganese Iron Boron,
Molybdenum and zinc.
Fertilizers
• Any deficiency from the above nutrients characterized by
certain symptoms.
• Various parts of plants are used in pharmaceutical industry
for their active constituents. Some of the nutrients are
responsible for growth of particular parts of the plant body
and hence their usefulness and deficiency need to be
described systematically.
• NPK fertilizer:
• Manure:
• Bio-fertilizer:
Propagation
Medicinal plants can be propagated by using following
methods as applicable to non-medicinal plants.
These are:
A. Sexual method
B. Asexual Method
C. Aseptic methods of Micro propagation
D. By inoculation for fermentation.
A) Sexual Method: In this method plants are raised from

seeds and such plants are known as seedlings.
A) Asexual method:
i. Vegetative propagation: In case of asexual method of
vegetative propagation, the vegetative part of a plant is
placed in such an environmental that it develops into a
new plant.
• From stem, bulb(squill, garlic)
• From corm (colchicom)
• From tuber (potato, aconite)
• From root, rhizome (ginger, turmeric)
• From runner(peppermint)
• From sucker (mint, chrysinthemum)
• From offset (aloe) or stolon (liquorice) etc.
ii. Grafting or budding: Stock with scion

C) Aseptic methods of Micro propagation:
• It is novel method for the propagation of medicinal
plants.
• In micro propagation the plants are developed in an
artificial medium (provided with nutritional and
hormonal requirements) under aseptic condition
from very fine pieces of plants like single cell, callus,
seed, embryos, root tips, shoot tips, pollen grains
etc.
D) By inoculation for fermentation:

• This process applies particularly to the production
of moulds and the bacteria and is extensively used
in the manufacturer of antibiotics.
Collection
Crude drugs are collected suitably when they contain maximum
concentration of active constituents.
Various factors for collection of crude drugs are:
(I) Time of collection: Time of collection plays an important role for the
crude the collection. E.g. Jasmine at morning time (in low temperature).
(II) Season of collection: The season at which crude drug is collected is
usually a matter of considerable important, since the amount and some
times the nature of the active constituents not constant though out the
years.
• Barks are generally collected in the spring or in the early summer
when the cambium is active as it is easy to detach from the stem,
some time they are collected in autumn or in rainy season (cinnamon)
• The roots are collected in spring before vegetative process stops.
(III) Ontogenic variation (age factor): The age of the plant is also a factor
for consideration as ontogenic variation. E.g. Turpentine oleo-resin and
balsam of peru are collected when the plants is about 8-10 years old.
(IV) Other conditions:
• The drugs which are leaf and the flowering tips of the plants, are
collected just before they reach their flowering stage or maturity;
example - Senna, digitalis, etc.
• Flowers are collected just before pollination or before their full
expansion e.g. clove.
• Fruits are collected when these are fully grown and ripe, but in some
case when partially ripe fruits are collected as per requirement.
Generally fruits are collected just before their dehiscence.
• Leaves, flower fruits should not be collected when covered with dew,
and rain.
• Rhizomes are collected when they store ample of reserve food
material and also contain maximum content of chemical
constituents.
• Unorganized drugs are preferably collected in dry weather
• Drugs such as resins gums, lattices are collected as soon as they
are out of the plants e.g. acacia gum is collected when it is
sufficiently hard after oozing out from the bark.
Drying
Drying consists of removal of moisture content of the crude
drugs. The slicing and cutting into smaller pieces is done to
enhance drying.
Drying should be carried out as soon as possible after
collection (necessary in case of glycoside containing drugs).
Drying facilitates to:
– Improve the quality and make the drug resistance to the
growth of microorganism. Before marketing a crude drug.
It is necessary to process it properly.
– Preserve it for a longer time and also to acquire better
pharmaceutical elegance.
– Process of grinding.
– Prevents partially the enzymatic reaction. If enzymatic
reaction to be bounced, drying
• Depending upon the type of chemical constituents a method of drying
can be adopted which are as the following types;
(I) Natural Drying (Sun Drying):
(A) Drying under sun: If the contents of the drug are quite stable to the
temperature and sun light can be dried directly in sunshine (gum acacia,
seeds and fruits).
(B) Drying in shade: If natural colour of the drug and the volatile
principles of the drug are to be retained then it is dried under shed, in
dry weather.
(II) Artificial drying: Drying by artificial means includes, drying of a drug in

(A) Oven dryer (Tray dryer): Non volatile drugs such as belladonna,
cinchona bark etc.
(B) Vacuum dryer: Drugs which are sensitive to higher temperature e. g.
digitals, tannic acid etc.
(C) Spray dryer: Drugs which are highly sensitive to atmospheric
condition such as leaves herbs, flowers are dried at 20’ to 40’ c.
When delicate structures are over dried, they become brittle and tend to
break in transit.
Packing
• The morphological and economical nature of the drugs, their
ultimate use, the effects of climatic conditions during
transportation and storage should be taken into
consideration while packing the drugs.
• Drugs are packed in sacks, bales wooden cases, card boards,
boxes and paper bags.
– Aloe is packed in goat skin.
– Colophony and tolu balsam in kerosene tins.
– Asafoetida in well closed containers.
– Cod liver oil in light resistant air tight container.
– Senna, vinca are pressed and baled.

Storage
• Usually drugs are stored in dark place to prevent light.
• Drugs sometimes absorb moisture in the time of storage, are often
termed as air dry and for these, air tight containers are recommended;
example - air in the container is released before storing fixed oils.
• Drugs are always liable to attack of insets and other pests, so they
should be frequently examined during storage and any showing
mould, worm or insect should be either rejected or treated.
• Fungal growth can be prevented by using fumigants.
• Temperature control should be done during storage.
• Volatile oils in well filled and sealed containers.
TYPES OF PESTS
1. Fungi:Ascoclyta
atropae (necrosis of
leaves).
2. Viruses: Mosaic virus,
ring spot virus
3. Weds:
4. Insects: Flea beetle,

Heliothis armigera.
5. Non insects:
i. Vertebrates - Rat,
Monkeys, birds
ii. Invertebrates -
Nematodes, Mites.
1. Rodenticides: Red Squill, Strychnine.
2. Insecticides: Rotenoid, Peyrethroids.
3. Acaricides (Miticides): Tetradion.
4. Fungicides : Antibiotics, Chlorophenols.
5. Herbicides: 2, 4-dichlorophenoxy acetic acid.

Insecticides are
applied to vegetative
parts for protective or
eradicant activity.
Act either by:
• Nerotoxication
• Inhibition of acetyl
cholinesterase
• Inhibition of
photosynthesis
• Inhibition of
chlorophyll
synthesis
• Hormone analogs
Classification
• These are used in the

form of aerosol, spray,
solution, suspensions,
fine dust and
fumigation etc.
DESIRED CHARACTERS:
1. Should non toxic, non Injurious to medicinal plants and
human being.
2. Should be selective in action and highly toxic to the insects
in low concentration.
3. Pesticides should be stable under ordinary condition of
storage.
4. It should not be inflammable, corrosive.
5. Should be free from obnoxious odour.
6. Should be non–cumulative in soil.
Source Macroscopy Chemical Use
Constituent
Pyrethrum: Insect The closed Pyrethrum owes its Insectici
flower flowers heads insecticidal property to -dal
Biological Source: are about 6-9 Easters. (Contact
Pyrethrum flowers mm. in diameter 1. Esters of Poison).
Chrysanthemum acid.
are the dried flower and the open
• Pyrethrum-I
heads of ones about 9-12
• Jasmoline – I
ysanthemum mm in diameter.
2. Esters of purethric
erarifolium
Colour–Cream to acid
(Family: Composite).
• Pyrethrin-II
straw coloured,
Geographical Source: • Jasmoline – II
Odour- Aromatic , • Cinerine-II are.
Indigenous to
Balkans. It also contains
Neem-
Leaves: Imparipinnate,, 1. Isoprenoids or Nimbine:
alternate opposite. terpenoids Antiviral,
Shape: ovate to A) Di- terpenoids Salanin:
(Margosa, Indian lilac) laceolate. Antifeedant.
B) Tri-terpenoids
Margin-serrate Nimbidine:
Biological Source: It Colour- green, 2. Norterpenoids Antiviral,
consists of dried Taste - bitter. A) Nor-tri- Azadirechtin:
leaves, root bark, Bark: Moderately thick, terpenoids: Insect
stem bark and other Rough; colour- starchy repellant,
white, laminated. 3. Limonoids - Meliantrol:
parts of Azadiracta
Odour-characteristic, Bitter principles Antifeedant,
indica (Meliaceae). Taste-bitter. (Nimbine, nibibin, Nimbosterol:
Large evergreen tree Flower: Numerous in nimbidine, salanin, Insecticidal,
of height 12 to 18 mt. axillary panicles. Colour- Margolone &
azadirechtin,
with a straight bole white or pale yellow. margolonone:
Fruit: Drupaceous, nimbolin-B etc.). Antibacterial
and long spreading
branches. colour- green turning 4. Other
yellow on ripening. constituents: Other uses:
Geographical Source: Seed: Single seed. spermicidal,
meliantrol,
Found through out Neem oil (margosa oil): antimalarial,
Fixed oils expressed quercetin, anti-
the drier parts
from seed carnels. myrecetin, Inflammatory
(Pakistan,
Colour- pale yellow. margolone, and anti-
Bangladesh, Srilanka,
Odour- garlic like, margolonone, and tumor etc.
Thailand, Malaysia, Taste- bitter. nimatone,
Source Chemical Constituents Use
Derris and Lonchocarpus

The roots of many species of Insecticidal properties Insecticidal
Derris Derris
- elliptica which are usually but not (Contact poison).
(Leguminosae) and dried roots invariably due to the Powder of one or
Lonchocarpus
of utilize, presence of Rotenone. mixture of both.
(Leguminosae).
Nicotine
Nicotine is the characteristics In addition to Nicotine Insecticidal.
alkaloid obtained from the alkaloids the
leaves of genus Nicotiana, homologous, Anabasine
Nicotiana tobacum (solanaceae) is also having

and is prepared commercially insecticidal properties.
from waste materials of the

Source Chemical Use
Constituents
Strychnine
The occurrence of strychnine in Alkaloid - Strychnine The alkaloid Strychnine is a
Strychnus species, dried ripe seeds CNS stimulant and used
Strychnus
of nuxvomica as Rhodenticide.
(Loganiacea).
Ryania Insecticidal properties
It consists of roots and stem of Contain 0.6 to 0.2% of (control various

alkalioids (Ryanodine). lepidopterous larval,
Ryania speciosa (Flacourtriaceae).
eurepean corn cores).
Citronella oil
Deep yellow volatile oil obtained by Geranial, Citronellal. Citronellal is Insect
distillation of Cymbopogon- repellant.
nardus ( Gramineae).
Source Chemical Use
Constituents
Sevadilla Seed Seeds contain about 2-4% The powdered seeds and
Sabadilla or
Cevadilla of mixed alkaloids: preparations of vertrine
consists of the Cevadine, Veratridine, are used as a dust or
dried ripe seeds
of Sabadine, Sabadiline spray to control thrips and
choenocaulon known as veratrine various true bugs which
fficinale
(Liliaceal). alkaloids. attack vegetables.
Red Squill In addition to other cardio Unlike other mammals,
Red Squill and
white squill are active glycosides the bulb rodent donot regurgitate
both varieties of of the red squill also the squill bulb, and death
Drimia maritima
(Liliacea). contains other glycosides follows convulsion and
like: scillirodise Scilliroside respiratory failure.
Plant Growth Regulators
• Plant growth regulators are the organic compounds, other
than nutrients which affect the morphological structure
and physiological process of plants in low concentrations.
• Plant growth regulators include both the native
(endogenous) and the synthetic (exogenous) substances.
• Phytohormones or plant hormones are naturally occurring
plant growth regulators.
• Auxins, gibberlines, cytokinins, absicic acid and ethylene
are five major plant growth regulators.
• Auxins, gibberlines, cytokinins, are plant growth
enhancers
• Absicic acid and ethylene are plant growth inhibbitors.
• Auxins, gibberlines and cytokinins are multiple forms of
endogenous forms of plant growth regulators.
• Plant growth regulators established their status
specifically in enhancing the size of the plant and
production of secondary metabolites used as drug.
Auxins
• Auxin is a general term used to indicate substances that promote
elongation of coleptile tissue.
• These have similar properties to Indole-3-acetic acid (IAA) which is
a major auxin of plants and found particularly in actively growing
tissues.
• Other natural auxins are:
o Indole-3-acetonitrile,
o 4-chloro indole-3-acetic acid (IAA)
o Phenyl acetic acid.
• The synthetic auxins are: Physiological effects:
o Indole-3-butyric acid (IBA) • Cell elongation
o α-napthyl acetic acid (NAA) • Root initiation
o 4-dichloro phenoxy acetic acid (2,4-D)• Prevention of abscission
• Induction of
o 2-napthyl oxy acetic acid (NOA)
parthenocarpy
o 1-napthyl acetamide (NAD) etc. • Stimulates respiration
• Callus formation
Mechanism of action: The proposed
mechanism of action of IAA is it’s interaction
with one or more components of biochemical
synthesis involved in the protein synthesis.
Practical uses:
• In low concentration it accelerates the
rooting of woody and herbaceous cuttings.
• In higher concentration act as herbicide and
weed killer.
Pharmaceutical applications: When seedlings

ofMentha piperita are treated with auxins,
40% increase in volatile content is observed.
GIBBERELLINS
• They are a class of endogenous plant growth regulators.
• At present over 50 gibberellins are known.
• About 40 of them are present in green plants, while other are
present in some fungi.
• They are present in different organic and tissue like roots
shoots, buds, leaves floral apices root nodules, fruits and
callus tissues.
• Gibberellins are synthesized in leaves and they accumulate
in relatively large quantities in the immature seeds and
fruits of some plants.
• Gibberellins are referred to as GA1. GA2, GA 3, GA4, GA7 and
GA9.
• GA3 is termed as gibberlic acid.
• All of them are the derivatives of gibbane ring skeleton.
• GA has not yet synthesized but can be produced by large
Functions: The commercial formulations of gibberellins are used currently for
promoting:
• vegetative and fruit growth
• breaking dormancy, flower initiation
• induction of parthenocarpy
• internode elongation.
Pharmaceutical applications: The applications of gibberellins are extended to

various medicinal plants:
• The use of gibberellins in lower dose has shown increased yield of digitalis
per shoot. The hormone tried with leaf and root culture of digitalis, showed
higher production of digoxin.
• It is observed that GA treatment can cause an increase in height of castor
plant up to 5 times but does not show any change in fixed oil content.
• The treatment significantly causes reduction in alkaloid content in vinca,
datura.
Mechanism of action:
• The growth effect of gibberellins arises by cell elongation in the sub-apical
meristem region where young internodes are developing.
• The effect of gibberellins and Auxins appear complimentary.
• The full stimulation of elongation by either hormone necessitating an
adequate presence of the other.
CYTOKININS
• Auxins and gibberellins are concerned largely with cell
enlargement and though they influence cell
multiplication process
• Cytokinines have a more specific effect on cell division
(Cytokinesis).
• Cytokinines are found in young and actively tissues like
embryos, seed lings and apical meristems.
• Natural occurring cytokinins are zeatine (N -dimethyl
6
amino Purin) and (N6-S –Isopentyl aminopurine.

2
• The synthetic cytokinins are kinetin, adenine, 6-benzyl

adenine, benzimidazole and N, N diphenyl urea.
1
• In plant metabolism, it is proposed that some t-RNA
contain cytokinin like activity.
• They have an action on some enzymes responsible for
formation of certain amino acids.
• Functions:
– Promotion of cell division.
– Participation in orderly development of embryos during
seed development influencing the expansion of cells in
leaf discs and cotyledons.
– Delaying senescence (the ageing process of the leaves
usually accompanies with loss of chlorophyll i.e. yellowing
and rapid breakdown of protein).
– Cell enlargement (Induces cell enlargement).
– Initiation of infrastructure cambium.
– Cause morphogenetic change.
– Dormancy of seeds.
– Counteraction of apical dominance.
– Cytokinins have been much employed in tissue culture
work in which they are used to promote the formation of
adventitious buds and shorts from undifferentiated cells.
Pharmaceutical applications:
• Cytokinins are reported to increase marginally
sennoside content in Tirnnevally senna leaves and also
enhance the dry weight of shoots.
• In opium they cause formation of elongated capsule
and reduced alkaloid content.
• Kinetins are reported to play a role in nucleic acid
metabolism and protein synthesis.
ETHYLENE (PLANT GROWTH INHIBITORS)
• It is a simple organic molecule present in the forms of volatile gas.
• Shows profound physiological effects on plants.
• It is present in ripening fruit, flower, stem, roots, tubers and seeds.
• It is present in very less quantity in plant (normally about 0.1 PPM).
• Possibly its quantity is increased in local areas during the time of
growth and development.
• Ethylene produced by one plant may influence the growths of the
other plants near to it.
Functions:
• Fruit ripening. Pharmaceutical applications:
• Stimulate leaf abscission. • At low concentration has
• Causes inhibition of root growth. been shown to increase the
• Stimulates formation of adventitious sennoside concentration in
roots. Cassia angustifolia.
• Stimulates production of
• Post harvest maturation of freshly fruits.
• It stimulates fading of some flowers. the stress compounds
• It stimulates epinasty of leaves. (phytuberin and phytuberol)
in tobacco leaves.
ABSICIC ACID (PLANT GROWTH INHIBITORS)
• A diffusible abscission accelerating substances
was found by Osborne in senescent leaves.
• Carns et. al. isolated several abscission
accelerating substances from cotton plants and
named as absicisin-I and abscisin- II (ABA).
• In an inhibitory way ABA interacts with other
plant growth substances in many seeds and
helps in seed dormancy.
• ABA concentration are found to be enhanced in
stress condition like mineral deficiency, injury,
draught and flooding.
• ABA serves an important as potential anti-
transpirant by closing the stomata when applied
to leaves.
HYBRIDIZATION
“The process through which the hybrid is produced is called
hybridization.”
•A hybrid is an organism which results from the crossing of two

species or verities differing at least in one set of characters.
•The resultant hybrids are:
omonohybrid (have one pair of different character)
odi-hybrids (two pairs of different characters)
opolyhybrids more than two pairs of characters.
•Hybrids helps in inducing the favorable characters of other varieties
or species
•Some times hybrids produce new and favorable characters in a
single variety which are not present in both the parent.
Example: The hybridization of Withania somnifera Israeli
chemo type - II and W. sominefera South African chemo type
has lead to formation of a new hybrid which contains 3 new
withanoloids.
•A recent development in hybridization is through the medium of
tissue culture (protoplasts fusion through protoplast cultures) are
employed for this purpose.
MUTATION
“Mutation is represented as variation in the character of the
species.”
• This is significant for the medicinal plants.
• The changes caused due to mutations included:
o morphological and anatomical changes
o changes in the chemical composition of the plants; this is
significant for the medicinal plants.
o Change in the yield
o Mutations may cause building resistance of a medicinal
plant towards certain diseases and form resistance. But in
all cases the plant may become susceptible to climate
conditions and certain other diseases. These effects are
to be eliminated by breeding and selection.
Cause of Mutation
1. Due to the environmental changes:
o The variations are observed but the original
traits are restored when the changes in
environment are withdrawn or disappeared.
o This type of change and re-storage is not
heritable and also not built into the
genotype.
o They are termed only as phenotypic
variations and commonly called as
modification.
2. Due to the changes in hereditary constitution.

Mutation is distinguished Spontaneous mutation:
into two types: o Mutation which occurs due
to some unknown reason
1. Chromosomal Mutation: from nature.
o This leads to change o This has been observed in
in amount or position some plants, bacteria
of genetic material. viruses etc.
o The chromosomal, Artificial mutations:
mutation is also called o
chromosomal Mutation can also be
aberration. induced by artificial means.
o Those which are induced
1. Point mutation: artificially in the living
o The changes with a organism by exposing them
gene or cistron of the to mutagens (abnormal
DNA molecule. environment)
o It is permanent and o Mutagens (mutagenic
heritable. agents): Radiations,
chemicals etc.
Mutagens or mutagenic agents:
1. Radiation mutagens: The electromagnetic waves of short

wave length of U.V. light, X-ray, gamma ray, α-ray and β- ray
are the radiation mutagens.
• The X- ray and gamma rays are called ionizing radiation and
also include α particle and β rays.
• Ionizing radiation causes:
o Water molecule in a biological system releases into H+
and OH - radicals. H+ reacts with O2 and produces
Hydroperoxils (HO2) radical.
o Both these radicals viz. hydroperoxyl and hydroxyl are
potent oxidizing agents.
o When chromosomes and their DNA are struck by such
radicals, they react due to which sugar phosphate part of
DNA may be impaired leading to chromosomal mutation
like breaks, deletions, additions, inversions and
Mutagens or mutagenic agents:
2. Chemical mutagens:
i. Nitrogen mustard, formaldehyde, nitrous
acid and ethyl ethane sulphonate alter
the chemical constituents of DNA bases
and cause transitional substitution in
DNA.
ii. 2-aminopurine, urethan, 5-bromouracil,
caffeine, phenol and certain other
carcinogens act as base analogs and
bring out copy error mutation in DNA.
• The artificial production of mutation in

medicinal plants is an important mile stone in
the development of cultivation technology.
Chemical races
“Chemical races are group plants which have similar
morphological characters but differs in type and
quantity of chemical constituents”
Example: The hybridization of Withania somnifera

Israeli chemo type - II and W. sominefera South
African chemo type has lead to formation of a
new hybrid which contains three new withanoloids.
GABA AND OTHER TYPES OF
NEUROTRANSMITTER
INTRODUCTION TO SECONDARY METABOLITES:
IDEFINITION, CLASSIFICATION, PROPERTIES AND TEST FOR
IDENTIFICATION OF ALKALOIDS, GLYCOSIDES, FLAVONOIDS, TANNINS,
VOLATILE OIL AND RESINS
Dr. I. P. Padhy
• Phytochemicals are chemical compounds that contains
naturally in plants.
• The prefix “phyto” generated from Greek word which
means plant.
• Phytochemicals are of two types:
1. Primary phytochemicals or primary metabolites
2. Secondary phytochemicals or secondary metabolites
Alkaloids (alkal = alkali like + Oids = group of compounds)
“Typical alkaloids are organic plant products of natural

origin which are basic in nature and contain one or more
nitrogen atoms in its heterocyclic ring”.
These are optically active and possess specific physiological

action on human or animal body, when administered in small
quantities.
Properties
• Basic, crystalline substances which unite with acids to form salts.
• Generally solid, exception is Nicotine (liquid).
• Generally white coloured, exception is Barberin (yellow).
• Odourless.
• Bitter or intensely bitter taste.
• Optically active
• Free bases are frequently sparingly soluble in water, but soluble in organic solvents,
with salts theses being usually soluble in water but sparingly soluble in organic
solvents.
Example- strychnine Hcl. is much more soluble in water than is strychnine base.
Exception - Caffeine (base) is readily extracted from tea leaves with water and
alkalodal salts like quinine sulphate is only soluble to the extent of 1 part in 1000
parts of water.
• Nitrogen atoms of alkaloids are may be of primary (Mescaline), secondary
(Ephedrine), tertiary (Atropine) of nature or quaternary ammonium compounds
(Tubarcuranine Cl.)
Effects of Alkaloids on Humans
• High biological activity
• Produce varying degrees of physiological and psychological
responses - largely by interfering with neurotransmitters
– others interfere with membrane transport, protein synthesis or other
processes
• In large doses - highly toxic – may be fatal
• In small doses, many have therapeutic value
• Generally used as muscle relaxants, tranquilizers, pain killers,
mind altering drugs, chemotherapy.
Types of alkaloids
• True alkaloids: Present in plants as salts of organic acids.
• Proto alkaloids or amino alkaloids: Lack one or more properties of
typical alkaloids like nitrogen atom is not present in its heterocyclic
ring. Example: Mescaline.
• Pseudo alkaloids: Steroidal or terpenoidal alkaloids (not derived
from amino acids). Example: Caffeine.
• Other alkaloids: These are not conforming to the general
definition, are those synthetic compounds, not found in plants, but
very closely related to the natural alkaloids. Example:
Homoatropine.
Source
• There are about 6000 alkaloidal compounds identified yet.
• Present in 15% of all vascular plants from 150 plant families.
• Nearly 300 alkaloids belonging to more then 24 classes, are known
to occur in the skin of amphibians. Some reptiles and mammals
also possess alkaloids (The Alkaloids, 1993, 43, 119).
• Synthetic alkaloids.
ALKALOIDS BIO-SYNTHESIS
• Most alkaloids are synthesized from a few common amino acids
(tyrosine, tryptophan, ornithine or argenine, aspartic acid and
lysine).
• Nicotinic acid is the precursor for part of nicotine.
• Purine is the precursor for caffeine.
• Some alkaloids synthesized from terpenes - along the mevalonic
acid pathway.
Classification
Taxonomic method for classification of alkaloids: This method of
classification is based on the taxonomical position of plant which
contains the alkaloidal compounds.
• Solanaceous alkaloids. Example: Atropine.
• Rubiaceous alkaloids. Example: Quinine.
Biosynthetic method for classification of alkaloids: This method of
classification is based on their precursors.
• Alkaloids synthesized from Tryptophane. Example: Indole alkaloids.
• Alkaloids synthesized from Ornithine. Example: tropane alkaloids.
Pharmacological method for classification of alkaloids: This method of
classification is based on the pharmacological action.
• Alkaloids acting as analgesic. Example: Morphine.
• Alkaloids acting as antitussive. Example: Codine.
• Alkaloids acting as antimalarial. Example: Quinine, chinchonine.
Classification
Chemical method for classification of alkaloids: Generally classified by the
predominant ring structure and/or carbon skeleton.here are two broad divisions.
I. Non heterocyclic alkaloids.
II. II. Heterocyclic or typical alkaloids.
1. Pyrrole and pyrolidine alkaloids. Example: Hygrine.
2. Pyrolizidine alkaloids. Example: Symphitine.
3. Pyridine and piperidine alkaloids. Example: Nicotine.
4. Tropane(Piperidine / n-methyl-pyrolidine) alkaloids. Example: Atropine, cocaine.
5. Quinoline alkaloids. Example: Quinine, chinchonine, cinchonidine etc.
6. Isoquinoline alkaloids. Example: Morphine, codine, papaverine etc.
7. Aporphine (Reduced Isoquinoline / naphthalene) alkaloids. Example: Boldine.
8. Norlupinane alkaloids. Example: Spatein.
9. Indole or benzopyrrrole alkaloids. Example: Ergotamine, reserpine, vinblastin,
vincristin, strychnine, brucine etc.
10. Indolizidine alkaloids. Example: Castanospermine.
11. Imidazole alkaloids. Example: Pilocarpine.
12. Purine alkaloids. Example: Caffine.
13. Steroidal alkaloids (some combined as glycosides). Example: Solanidine.
14. Terpenoid alkaloids. Example: Aconitine.
Chemical tests for alkaloids
Tests with following reagents (precipitation tests) are carried out to
detect the presence of alkaloids.
Alkaloidal extracts (organic solvent) are treated with dilute acid and
aqueous portion is separated. To few ml. of this solution, in
sepatrte test tubes, a drop of following reagents for general tests
are added from the side wall.
General tests:
1) Mayer’s reagent test: (Potassium mercuric chloride) → Cream
coloured precipitate.
2) Dragendroff’s reagent test: (Potassium bismuth iodide) → Reddish
brown coloured precipitate.
3) Wagner’s reagent test: (Iodine in potassium iodide) → Brown
4) Hager’s reagent test: (Saturated solution of picric acid) → Yellow
5) Kraut’s reagent test (modified dragendroff’s reagent) → Reddish
Chemical Tests for Alkaloids
Specific tests:
Murexide test for purine alkaloids: To 3 - 4 ml of test solution, add
small amount of potassium chlorate and a drop of HCl. Evaporate
to dryness and expose to ammonia vapour → purple colour is
observed.
Vitali’s test for tropane alkaloids: Mix a drop of fuming nitric acid
with solid alkaloidal sample (as less as 1µg.) and evaporate to
dryness at 100°C. Add 0.5 ml of 3% solution of alcoholic KOH
solution to the residue → A bright purple colour which changes to
red and subsequently fades to colourless.
Thalleioquin test for quinoline alkaloids: Few drops of bromine
water + 2-3 ml of a weakly acidic solution of a quinine salt + 0.5 to
1 ml of strong ammonia → a characteristic emerald green colour
is produced.
Van Urk reagent (Ehrlich reagent) test for indole alkaloids:
Indole alkaloids + Van Urk reagent (p-
dimethylaminobenzaldehyde) → a characteristic deep blue
colour.
GLYCOSIDES
“Glycosides may be defined as

the organic compounds from
plants or animals which is
comprising of a sugar portion
linked to a non sugar moiety in
a particular manner (through
glycosidic bond) and on
enzymatic or acid hydrolysis
produces the same.”
Hydrolysis
• Sugar moiety of a glycoside is known as glycone or genin.

• Non sugar moiety of a glycoside is called the aglycone or aglycogenin.
• Sugars in glycosides are most commonly D-glucose, others are
galactose, mannose, rhamnose, digitoxose or cymarose etc.
• Number of sugar moieties in glycosides are one or more.
• The linkage between glycone and aglycone is a hemiacetal formed
by reducing group (usually aldehyde or keto group) of the sugar
and an alcoholic or phenolic hydroxyl group of the aglycon
(glycosidic linkage).
Properties:
• Crystalline or amorphous substances.
• Soluble in water, dilute alcohol, insoluble in organic solvents like
chloroform and ether.
• Optically active.
• Easily hydrolyzed by water, mineral acids and enzymes.
• Do not reduce Fehling's solution until hydrolyzed.
General Uses:
• Glycosides are used as cardio tonic, purgative, anti rheumatic and
expectorant etc.
• Believed to participate in growth regulation and protection of the plants.
General Chemical Tests:
1. Sugar content test: Determine free sugar content of the extract.
Hydrolyse the extract with mineral acid ( dilute HCl / dilute H2SO4 ) and
again determine the total sugar content of the hydrolised extract.
Increase in sugar content indicates presence of glycoside in the extract.
2. Baljet test: A thick section of crude drug shows yellow to orange colour
with sodium picrate.
Classification
I) Based on the types of linkage:
• C - Glycoside: Aloe.
Glycone – OH + HC – aglycone → Glycone –C-glycone + H20
• O - Glycoside: Senna
Glycone –OH + HO- aglycone → glycone-O-aglycone + H20
• S – Glycoside (Isothocyanate glycosides): Sinigrin
Glycone – OH+ HS –aglycone → Glycone -S-aglycone + H20
• N - Glycoside: Nucleosides
Glycone –OH + HN –aglycone → Glycone –N-aglycone + H20

……Classification
II) According to the sugar moiety:
• Glycosides with glucose.
• Glycosides with rhamnose.
• Glycosides with ribose.
• Glycosides with digitoxose etc.
III) According to therapeutic action:

• Glycosides as cathartics
• Glycosides as analgesics
• Glycosides as expectorant
• Glycosides as cardio tonics etc.
Chemical Classification (according to chemical nature of aglycone moiety):
1. Anthraquinone glycoside: Senna
2. Cardiac glycosides: Digitalis
3. Saponin glycosides: Liquorice
4. Cyanogenetic or cyanophoretic glycosides: Bitter almond
5. Isothiocyanate glycosides: Black mustard
6. Flavonol glycosides; example: Citrus fruits
7. Coumarin and furauno-coumarin glycosides: Tonka bean camphor
8. Aldehyde glycosides: Vanilla
9. Phenolic glycosides: Bear berry
10. Steroidal glycosides: Solanum
11. Glycosidal biters or miscellaneous glycosides: Saffron
Gold beater’s skin test for condensed tannins:
• An intestinal membrane of ox or sheep is treated with HCl
• Rinsed with distilled water
• Treated with tannin solution for 05 minutes
• Washed with distilled water
• Treated with ferrous sulphate solution → a brown or black

colour is developed on the skin due to tannin.
Extraction of Tannins: Tannins can be Extracted using

solvents like alcohol or acetone in a soxhelator.
Resins and Resin Combination
“Resins are amorphous products of complex chemical nature and are

mixtures of essential oils, oxygenated products of terpens and
carboxylic acids containing large no of carbon atoms, found as
exudations from the trunk of various trees.”
❖ In many instances resin in plants are formed in special passages

or tubes called resin ducts, schizogenous and schizolysigenous
glands as a product of metabolism, thus a single incision may
drain the resin from a considerable area of the plant.
Properties Extraction Methods:
• Heavier than water 1. Extracted with alcohol and
• Insoluble in water; soluble in then precipitated with
alcohol, volatile oils, fixed oils water; Example – Ipomoea.
and other non-polar organic 2. Distillation for separation of
solvents oils; Example – Copaiba,
• Non conductive to electricity Colophony.
• These are Transparent or • Obtained as plant exudates
translucent solids, semisolids or by incision; Example –
liquid substances Myrrh, Asafetida etc.
• These are hard, when heated • Heating Plant parts
often and ultimately melt
• Resins burn with a characteristic
smoky flame
• Chemically they contain organic
acids, alcohols and esters
Classification
Depending upon the type of constituents, resins are classified as:
1) Acid Resins (resin acids or resinolic acids): These are of high molecular
weight and very complex compounds usually occur as free; example:
Abiatic acid, Commiphoric acid etc in Colophony.
2) Easter Resins: These consist mostly of resin alcohols combined with
aromatic acids like benzoic and cinnamic acids; other aromatic acids are
less frequently found, e.g. ferulic acid (in asafetida). There are two
principal kinds of resin alcohols in combination with these acids: e.g.
Benzoin, storax etc.
3) Resin alcohol (Resinols): The contents are the complex high molecular
weight alcohols: Balsams of peru.
4) Resin phenols (Resinotannols): Occurs combined with benzoic and
cinnamic acids eg. resinotannol in balsam of Tolu is
called toluresinotannol.
5) Resens (Insert compounds): The chemical nature of these compounds
are unknown. They are very stable, being unaffected by most chemical
reagents or by exposure to moisture and light,Gum Copal, sandarac etc.
Examples of resins and resins combinations
• Resins (consisting principally of resin and other esters, together
with free aromatic acids): Colophony and Cannabis etc.
• Oleoresins (Resins and volatile oils in homogenous mixture):
Copaiba, capcicum and ginger etc.
• Oleo-gum-resins (Homogenous mixtures of volatile oils, gums and
resins): Asafoetida and myrrh etc.
• Glycoresins (Made up of resins and sugars): Jalap and ipomoea etc.
• Balsams (resinous mixtures that contain large proportions of
cinnamic acid, benzoic acid or both or esters of these acids):
Balsam of Tolu, balsam of Peru etc.
VOLATILE OILS
Essential oil is a concentrated hydrophobic liquid
containing volatile aroma compounds obtained mainly
from plants and are volatile in steam.
Latin ‘essentia’ meaning a liquid easily changed to a gas.
Essential oils are also known as volatile oils, ethereal oils
or aetherolea.
“Volatile oils or essential oils are generally mixtures of
hydrocarbons and oxygenated compounds derived from
these hydrocarbons.”
• They differ entirely in both chemical and physical
properties from fixed oils.
Properties
• These are generally colorless liquids (exception chamomile which is
violet) or solids with pleasant smell.
• Volatilize in steam.
• Practically in-soluble in water.
• Soluble in alcohol, ether and other lipid solvents.
• Usually lighter then water.
• Possesses high refractive indices.
• Most of them are optically active.
• Many a times associated with other substances such as gums and
resins and them selves tend to resinfy on exposure to air.
• Terpeneless volatile oils are prepared by removing hydrocarbons and
undesired components using fractional distillation or any other
method, to make the oil stable.
• The odour and taste of volatile oils is mainly due to oxygenated
constituents, which are to some extent soluble in water but more
soluble in alcohol.
Source:
• Volatile oils are biosynthesized from phenyl propane
• Secreted from specially structures such as duct cells,
schizogenous glands, lysoschizogenous glands and glandular
trichomes etc.
• Volatile oils present either in entire plant or in part of the plant.
• Plants from families like labiatae, rutaceae, piperaceae,
zinziberaceae, umbeliferae, myrtaceae and lauraceae etc.
possesses volatile oils.
• There are about 100 commercially valuable volatile oils directly
derived from plants.
Uses: Volatile oils are generally used as flavoring agent,

carminative and antiseptic (oils with a high phenol content, e.g.
clove and thyme) etc.
Classification
On the basis of composition volatile oils are classified as following.
1. Terpenes and sesqui-terpenes (hydrocarbons) volatile oil
example: turpentine.
2. Alcohol volatile oil, example: peppermint, cardamom.
3. Aldehyde volatile oil, example: cinnamic aldehyde in cinnamon,
geraniol in citronella.
4. Ester volatile oil, example: lavender.
5. Ketone volatile oil, example: camphor, caraway.
6. Oxide volatile oil, example: eucalyptus.
7. Phenolic ether Alcohol volatile oil, example: anise, anethol in
fennel.
8. Phenol volatile oil, example: eugenol in clove.
9. Peroxide volatile oil, example: ascaridol in chenopodium.
10. Volatile oils which are non terpenoids and derivatives of
glycosides, example: glucosinolates in mustard and methyl
salicylate in gaultheria.
Chemical tests (volatile oils):
– Filter paper is not permanently stained with volatile oil.
– Solubility test: Volatile oils are soluble in 90% alcohol.
– Add alcoholic solution of sudan-III to the thin section of crude

drug containing volatile oil → Red colour formed by globules.
– Add a drop of tincture alkene to the thin section of crude drug

containing volatile oil → Red colour formed.
Classification
Dr. I. P. Padhy
INTRODUCTION
• In the domain of drugs and medicine, it is some what
contradictory to note that most of them are obtained by
synthesis or from terrestrial organisms.
• About 5 lakhs of species of marine organisms have been
reported from the oceans and seas from various parts of
the world.
• Some of these organisms are antimicrobial, antiviral,
antibiotics, enzyme inhibiters, anti-inflammatory,
neurophysiological, cardiovascular, anticancer/cytotoxic
agents.
• Many of the species contain toxic compounds.
• In western medicine some established products from
marine origin are; agar, alginic acid, spermaceti,
carragenin, cod liver oil, shark liver oil, halibut liver oil,
cephalosporines and protamine sulphate etc.
CLASSIFICATION:
ON THE BASIS OF ORGANISM ON THE BASIS OF THEIR ACTIVITY
1.Sea cucumber 1. Cardiovascular active compounds
(Stichopus japonicus) 2. Cytotoxic/ anticancer compounds
1.Brown algae 3. Antimicrobial compounds
(Dictyopteis zonaroids) 4. Antibiotics
1.Red algae 5. Anthelminitics
(Bonnemaisonia hemifera) (Stichopus japonicus)
1.Sponge (Verongia aerophoba) 1. Anticoagulants
2.Sea hare (Aplysia californica) 2. Insecticides
3.Gorgonian corals 3. Anti-inflamatary
(Eunicia mammosa) 4. Antispasmodics
1.Fungus 5. Vitamines
(Cephalosporium acremonium) 6. Miscellaneous and other
1.Marine bacterium principles
(Pseudomonas bromutillis) 7. Marine toxines
2.Marine bacterium
(Pseudomonas bromutillis)
CARDIOVASCULAR COMPOUNDS
Active Source Use
Compounds
Anthopleurines Anthopleurines are groups of peptides Cardio tonic effect (35
obtained from colenterates viz. times higher as compared
Anthropleura xanthogramica. to digoxin and less toxic).
Laminine Laminine is a marine algae Laminaria Hypotensive effect.
angustata.
Eptatretin Eptatretin is found in the aneural Potent cardiac stimulant
bronchial hearts of pacific hog fish, on mammalian
Eptatretus stoutii. myocardium.
d-octopamine D-octopamine is obtained from octopus Produces adrenergic and
macropus and Octopus vulgaris. cardiovascular response.
Saxitoxin Saxitoxin is obtained from Saxidomus Hypotensive effect.
giganteus.
Automonium Automonium is obtained from Verongia Produces adrenergic and
fistularis. cholinergic activity.
Spongisine Spongisine is a nucleoside obtained by Reduces both force and
extracting Caribbean sponge rate of contraction of
Cryptotethia crypta. heart.
ANTICANCER DRUGS
Active Source Use
Compounds
Ara-C Ara-C is 1- α – D- Arabinofuranosyl In acute
cytosine obtained from the myelogenous
Caribbean sponge viz. spongosine leukemia.
and spongouridine.
Cracin Cracin acetate is a cyclic di-terpene Cytotoxic
acetate obtained from soft corals Caribbean
gorganian.
Asperidiol It is a gorgonian coral. Cytotoxic
Aplisistatin It is obtained from sea hare Aplisia Anti leukemic.
angasi.
Halitoxin It is obtained from a sponge Antitumor
Helielona viridis. activity.
ANTIMICROBIAL AGENTS
Marine organism Antimicrobial principle
Sea cucumber (Stichopus Holotoxin-A, B & C
japonicus) (steroidal glycoside)-
antifungal.
Brown algae (Dictyopteis Zonarol and isozonarol.
zonaroids)
Red algae (Bonnemaisonia Tetrabroheptanone.
hemifera)
Sponge (Verongia aerophoba) Aeroplysinin-1(+) and
Aeroplysinin-1(-)
Sea hare (Aplysia californica) Debromol aurenterol.
Gorgonian corals (Eunicia Eunicin.
mammosa)
ANTIBIOTICS
Fungus (Cephalosporium Cephalosporin-C

acremonium)
Sponge (Ircinia strobillina) Ircinine-1
Marine bacterium 2, 4-dibromo-6-(3, 4, 5-
(Pseudomonas bromutillis) tribromopyrrol-2-4, 1)
phenol.
MISCELLANEOUS
Source Active Compounds Use
Eucheuma chondrus Laminarine Antilipimics
Plexaura homomalla Prostaglandins(PGA-2, Used in asthma, peptic
PGE-2) ulcer and in birth control.
Obtained from marine Ara-A Antiviral agent
sponge Tethya crpta
Rhodactis howesii Anticoagulant
Flustra foliaceae Flustramine A and B Muscle relaxant
(antispasmodics)
Cod liver oil, shark liver oil, Vitamine – A and D
halibut liver oil
Extract of various marine Vitamine – C, folic acid,
algae niacin and B-complex.
Sponge (Luffariella Monoalide Anti-inflammatory
variabilis)
Red algae (Digenia simplex) Anthelminitics
Dominic acid Chondria armata Anthelminitics
Dr. I. P. Padhy
INTRODUCTION
the world.
agents.
CLASSIFICATION:
2.Marine bacterium
Active Source Use
Compounds
angustata.
giganteus.
ANTICANCER DRUGS
Active Source Use
Compounds
and spongouridine.
gorganian.
angasi.
antifungal.
zonaroids)
hemifera)
Aeroplysinin-1(-)
mammosa)
ANTIBIOTICS

acremonium)
phenol.
MISCELLANEOUS
sponge Tethya crpta
(antispasmodics)
halibut liver oil
variabilis)
Neurohumoral Transmission in
C.N.S
1. The nerve action potential (AP) consists of a transient self-propagated
reversal of charge on the axonal membrane. The internal potential Ei goes
from a negative value, through zero potential, to a slightly positive value
primarily through increases in Na+ permeability and then returns to resting
values by an increase in K+ permeability. When the AP arrives at the
presynaptic terminal, it initiates release of the excitatory or inhibitory
transmitter. Depolarization at the nerve ending and entry of Ca2+ initiate
docking and then fusion of the synaptic vesicle with the membrane of the
nerve ending.
2. Combination of the excitatory transmitter with postsynaptic receptors
produces a localized depolarization, the excitatory postsynaptic potential
(EPSP), through an increase in permeability to cations, most notably Na+. An
inhibitory transmitter causes a selective increase in permeability to K+ or Cl–,
resulting in a localized hyperpolarization, the inhibitory postsynaptic
potential (IPSP).
3. The EPSP initiates a conducted AP in the postsynaptic neuron; this can be

prevented, however, by the hyperpolarization induced by a concurrent IPSP.
The transmitter is dissipated by enzymatic destruction, by reuptake into the
presynaptic terminal or adjacent glial cells, or by diffusion. Depolarization of
the postsynaptic membrane can permit Ca2+ entry if voltage-gated Ca2+
channels are present.
Parasympathomimetic
drugs
Presented By
Saroj kanta Bisoyi
Asst. Professor
RCPHS, BAM
Introduction
 The parasympathetic nervous system is also known as
cholinergic nervous system.
 Acetylcholine is the major neurotransmiter at post
ganglionic synapses of cholinergic nervous system.
 Acetylcholine was first reported by Reid Hunt and Taveau
in 1900.
 Dale (1914) reported two receptor Muscarnic and
Nicotinic Acetylcholine bind with the receptor shows
their physiological activity.
Biosynthetic pathway of Acetylcholine.
 The biosynthetic pathway of acetylcholine involve in

sequential six step.
1) Synthesis of Acetylcholine
2) Storage of Acetylcholine in vescicle.
3) Release of Acetylcholine
4) Binding of Acetylcholine to Receptor
5) Degradation of Acetylcholine.
6) Recycling of Choline.
Biosynthetic pathway of Acetylcholine.
(1) Synthesis of Acetylcholine
 Choline is transported from extracellular fluid into the
cytoplasm of cholinergic nerve by energy depedent
carrier cotransport system sodium channel.
 The uptake of Choline is the rate limiting step for
Acetylcholine and is block by the drug Hemicholinium.
 Choline Acetyl transferase enzyme catalysed the
reaction of choline with Acetyl Coenzyme-A to form an
ester in the cytosol.
 Acetyl Coenzyme-A is derived from the mitochondria
and is produced by the kerb cycle and fatty acid
oxidation.
(2) Storage of Acetylcholine
 Newly formed Acetylcholine is transported into
Cytosolic storage vesicle present in the presynaptic
nerve ending.
 It is present along with ATP and with calcium and

magnesium ion until it is release.
(3) Release of the Acetylcholine
 When an action potential propagated by the action of voltage
sensitive sodium chanel arrives at nerve ending that causes
influx of calcium into thee cell.
 Elevation of calcium level promote the fusion of synaptic
vesicle with the cell membrane and release of theire content
into the synatic space and it bind to either of two
postsynaptic receptor on the target cell.
 The postsynaptic cholinergic receptor on the surface of
effecttor organ are divided in to two classes.
(i) Muscarnic receptors
(ii)Nicotinic receptors.
4.Degradation of Acetylcholine
 Acetylcholine in the synapse can bind with
cholinergic receptor on postsynaptic membrane to
produced a responces.
 The free Acetylcholine Hydrolysed to choline and

acetate by enzyme Acetylcholine estarase.
(5)Degradation of Acetylcholine
(6) Recycle of Choline
 Choline may be recaptcured by sodium coupled high
affinity uptake system that transport the choline back
into the neuron.
 It is acetylted and stored until released by a
subsiquent action potential.
Actylcholine Receptors
 after releasing in the synapse Acetylcholine acts on
two receptors
these are
1. Muscarnic receptors (M-Receptors)
2. Nicotinic receptors (N-Receptors)
Muscarnic receptors
 Muscarnic receptors are G-Proteins coupled receptors
which activates other ion channels.
 Muscarinic receptors are more sensitive to Acetylcholine
than to nicotine receptors.
 Acetylcholine when bind to muscarnic receptor its causes
a confirmational changes in the receptors.
 Drug with muscarnic action potentially stimuates
muscarnic receptors on those tissue but at high
concentration they may shows some activity at Nicotinic
receptors.
Muscarnic receptors
 M1 receptors are found in gastric periatal cell
 M2 receptors on cardiac cell and smooth muscles
 M3 receptors on bladder,exocrine gland and smooth

muscle.
 M4 and M5 present in certain area of brain and
regulates other neurotransmiter.
 However till today M1,M2 and M3 are major one present
in effector cell and prejunctional nerve ending in CNS.
Mechanisam of Acetylcholine signal tranduction
 When M1 and M3 receptors activated the receptor
undergoes confirmational changes and interact with a G-
Proteine –Gq
Activated phospholipase
Hydrolysis of phosphatidylinositol(4,5)-bisphosphate.
Yield Diacylglycerol and ionsitol (1,4,5)triphosphate
Intracelular increase in calcium
Ths cation interact to stimulate or inhibit enzyme cause

hperpolarization contraction
Mechanisam of Acetylcholine
signal tranduction
 M2 subtype cardiac muscle stimulate a G- protein (Gi)
that inhibit adenyl cyclase
increase K+ condution to which heart respond to

dcrease in rate and force of contraction
Parasympathomimetic agents
 These are the agents or compounds which directly or
indirectly mimic the action of acetylcholine.
 These are the drugs which produced action similar to

that of Acetylcholine either by directly ineracting with
cholinergic receptors.
 Acetylcholine is a major neurotransmiter of

parasympathetic nervous system.
 Ssssssss
(1)The quaternary ammonium group
 Quaternary ammonium group is essential for activity.
 Replacement of nitrogen with sulphur(S) ,Arsenic(As) or selenum(Se)
produces less active compounds.
 Primary ,Secondary or quaternary amine are less active than
acetylcholine.
 The methyl group of quaternary ammonium group can be replaced
by alkyl group.
Ex. Dimethyl ethyl derivatives of acetylcholine is more active than
the parent drug.
 The replacement of more than one methyl group of
quaternary ammonium group leads to complete loss
of cholinergic activity.
(2) The ethylene bridge
 As the chain length increased from 2-carbon to more
than 2-carbon atom the activity is rapidly reduced.
 Branching in the chain i.e replacement of hydrogen
atom of ethylene bridge by methyl group leads to
equal or greater acetylcholine activity.
 The α or β-methyl substituted derivative affect selectivity
of receptors.
 Acetyl β-methylcholine act selectively on muscarnic
receptors which Acetyl α-methylcholine chloride has
greater nicotinic action than muscarnic activity.
(3) Acetyl group/Aceloxy group
 The higher homologus of methyl group propionyl or
butyryl group are less active than Acetylcholine.
 Ester of aromatic or higher molecular weight acid
possess cholinergic antagonist activity.
 Acetate group of Acetylcholine may be replaced by
carbamate.
Ex. Carbachol is more active and more stable than
parent drug.(Degradation is inhibited by enzyme Choline
esterase )
 Acetylcholine is directily acting drug act by bind to the
Muscarnic receptor and shows their action.
Uses
 Mostly used as a topical Opthalmic eye drop.
 It is used to redused the intraocular pressure associated with
cataract surgery , ridectomy and anterior surgery.
 It is also used as a vasodilator and cardiac depresant.
 It is also increases lachrymal ,salivary secretion.
Methacholine
 It is a Quaternary ammonium Parasympathomimetic agent with
muscarnic action of acetylcholine.
 It is more resistant to hydrolysis by the enzyme Choline esterase.
 Its shows prolong action.
Uses:
 Used in treatment of Glaucoma and reynaud syndrome.
 it is used in the treatment of asthma by inhalation route.
 Also used in the treatment of peripheral vascular disorder.

Carabachol
 IUPAC Name:
2-[(Aminocarbonyl)oxy]-N N
, N
, -trimethylethanaminium chloride
Carbachol
MOA
 It act by both muscarnic and nicotinic receptor and shows
action of acetylcholine.
 Its duration of action is increased due to inhibition degradation
of by the enzyme cholinesterase.
Uses
 It is used in the treatment of Glaucoma to reduce intraocular

pressure .
 It is also used for treatment of decreased gastrointenstinal
motility.
Bethanechol
 MOA
Bethanechol is a choline ester mainly act on muscarnic
receptor and shows their action.
 It is not inactivated by cholinesterase.
 It also have some nicotinic activity.
USES
 it is used in the relief of urinary retention and abdominal
distention after surgery.
 It also has prolong effect than Acetylcholine.
 MOA
Pilocarpine has mostly bind to M3 receptors in smooth muscle to
cause contraction in the eye and trachea.
 USES
 Treatment of Glaucoma.
 Treatment of dry eye or drymouth.
Anticholinesterases
 These are the agent which inhibt or protect acetylcholine from
hydrolysis.
 Acetylcholine accumulates in the cholinergic terminals and
increases the biological responces.
 These are of two types
1) Reversible inhibitores:
These drug binds reversiblly to the enzyme.
Ex- Physostigmine and Neostgmine
2)Irreversible inhibitors:
These drug produses irrevesible inactivation of Acetylcholine
estrase.
ex: Organophosphorus compounds.
They form a irreversible covalent bond to the active site of
the enzyme.
Ex: Parathion and Malathion
 MOA
Physostigmine is a reversible tertiary amine inhibitor of
enzyme Cholinestrase.
 Uses
Treatment of Glucoma
Also used parenterally for reversal the effects caused by
anticholinergic and tricyclic antidepresant.
Neostigmine
Neostgmine
 MOA
 Indirectly stimulates both muscarnic and nicotinic
receptors.
 It binds to the anionic and esteric site of cholinesterase
and block the activity of acetylcholinesterase.
Uses
 Treatment of myasthenia gravis.
 treatment of postoperative urinary retention.
 Used to lower intra-ocular pressure in the management
of guaucoma.
Pyridostigmine
 IUPAC Name:3-[(Dimethylamino carbonyl)oxy]-1-methyl pyridinium

bromide.
 Mechanism of action
 Pyridostigmine block acetyl choline esterase enzyme and inhibit the

destruction of release acetylcholine.
 Uses:
 It is mainly uesed in the treatment of myasthenia gravis.
 Also used as prophylaxis against the neurotransmiter effects of

nerve gas poisoning.
 MOA
 It is a reversible inhibitor of cholinesterase.
 Uses:
 It is mainly used in the treatment of Myasthenia gravis.
 also used in the treatment of snake bite.
 IUPAC Name: 1,2,3,4-Tetrahydroacridin-9-amine
 MOA:
 Tacrine is a centrally acting anticholinesterase.
 USES
 It is used in the treatment of mild to moderate
dementia in alzhemer’s disease.
 Also used as an analeptic agent to promote mental
alertness.
 MOA:
 It produses irreversible inactivation of acetylcholinesterase.
 USES:
 It is used as mitotic agent in treatment of glaucoma.
 Used in the treatment of organo phosphate toxicities.
.
 MOA:
 It is an irreversible acetylcholine inhibitors.
 It covalently bind to cholinesterase and permanatly
inactive the enzyme.
 Uses
 Treatment of chronic Glaucoma.
 MOA:
 It indirectly acts on the acetyl choline esterase enzyme.
 USES
 Used as insecticide in agriculture.
 Mostly applied by spraying to cotton rice and fruit tree.
Malathion
 IUPAC Name: Diethyl-2[(Dimethoxy phosphorothioyl) sulfanyl]

butanediote.
 MOA:
It act by binding to the serine residue on the cholinesterase
enzyme and irreversibly deactivates the enzyme choline esterase.
 Uses:
 At low doses (0.5%) it is used in the treatment of head lice and
body lice.
 Also used in the treatment of scabies.
 Mostly used in the treatment of insecticides.
Cholinesterase reactivator
 These are the drugs which causes reactivation of
choline esterase.
 These agent are react with alkylphosphorylated form of
cholinesterase to free the active part of the enzyme.
 Mostly these drug are used in the treatment of
organophosphorus poisoning.
 Ex: Pralidoxime chloride.
 IUPAC Name:2[(hydroxyimino)methyl]-1-methylpyridinium
chloride.
 MOA:
 It reactivates the Acetyl cholineserase enzyme rapidyl
by binding to the anionic site of enzyme and displace
the phosphate from the serine residue.
 Uses
 Mostly used in case of organophosphorus poisoning.
 Also used for the treatment of overdose by
anticholinergic drugs.
Dr. I. P. Padhy
means plant.


quantities.
Properties
• Odourless.
solvents.
parts of water.
(Tubarcuranine Cl.)
processes
Types of alkaloids
Homoatropine.
Source
lysine).
acid pathway.
Classification
Classification
General tests:
Specific tests:
observed.
is produced.
colour.
GLYCOSIDES

Hydrolysis

Properties:
General Uses:
expectorant etc.
Classification






smoky flame
Classification
VOLATILE OILS
or aetherolea.
Properties
soluble in alcohol.
Source:
trichomes etc.

Classification
fennel.


Classification
Dr. I. P. Padhy
INTRODUCTION
the world.
agents.
CLASSIFICATION:
2.Marine bacterium
Active Source Use
Compounds
angustata.
giganteus.
ANTICANCER DRUGS
Active Source Use
Compounds
and spongouridine.
gorganian.
angasi.
antifungal.
zonaroids)
hemifera)
Aeroplysinin-1(-)
mammosa)
ANTIBIOTICS

acremonium)
phenol.
MISCELLANEOUS
sponge Tethya crpta
(antispasmodics)
halibut liver oil
variabilis)
Dr. I. P. Padhy
INTRODUCTION
the world.
agents.
CLASSIFICATION:
2.Marine bacterium
Active Source Use
Compounds
angustata.
giganteus.
ANTICANCER DRUGS
Active Source Use
Compounds
and spongouridine.
gorganian.
angasi.
antifungal.
zonaroids)
hemifera)
Aeroplysinin-1(-)
mammosa)
ANTIBIOTICS

acremonium)
phenol.
MISCELLANEOUS
sponge Tethya crpta
(antispasmodics)
halibut liver oil
variabilis)
PLANT TISSUE CULTURE:
HISTORICAL DEVELOPMENT OF PLANT TISSUE CULTURE, TYPES OF CULTURES,
NUTRITIONAL REQUIREMENTS, GROWTH AND THEIR MAINTENANCE.
APPLICATIONS OF PLANT TISSUE CULTURE IN PHARMACOGNOSY.
Prof. I. P. Padhy
Tissue culture
“Refers to the aseptic technique of growing plant cells,
tissues or organs to a whole plant, in a sterile and
suitable environmental condition on a artificially
prepared nutrient medium”.
Concept
• “Totipotency” is the genetic
potential of a plant cell to
regenerate the whole organism
from a single cell.
Haberlandt- A German Botanist
1896
• “Plasticity” or adaptability by
Cultured isolated plant cells;
a plant to different was able to maintain the cell
in the medium but failed to
environmental conditions.
differentiate
Plants alter their metabolism, 1902
Haberlandt’s Hypothesis on
growth and development to
Totipotency
suit their environment.
History
• 1922- Robins (USA) and Kotte (Germany)- cultured plant
1920
root of tomato
• 1934- Nobercourt and Gautherate- Callus tissue culture

1930 • Discovery of auxins
• Miller and Skoog - Discovered Kinetin

1950 • 1954- Muir, Ricker and Hildebrandt – Suspension culture.
• Morel cultured orchids
• 1960- Hanging drop culture established

1960 • 1962- Murashige and Skoog developed MS Medium
• Murashige- cloned plants in-vitro
• Starting of genetic engineering
• 1972- Protoplast fusion carried out to hybridize two species
1970 of tobacco plant.
• Development of techniques to introduce foreign DNA into
plant cells
Technique of Tissue Culture
Stages
Preparation of appropriate culture media (nutrient media)

1
Growth of aseptic plant from surface sterilized explants

2 (seeds) and collection of the aseptic explants from this plant.
• Establishment of culture
• Multiplication - the explants gives rise to a callus
3 • Differentiation and organogenesis
Tissue Culture Media
Functions
Water
Mineral nutritional needs
Growth regulators
Provides Vitamins Functions

Organic compounds
Access to atmosphere for gas exchange
A base for plant growth
Serve as a dumping ground for plant metabolites

Components of Tissue Culture Media
Water Usually de-ionised double distilled (DD) water
Macro elements- Nitrogen, phosphorus,

potassium, magnesium, calcium and sulphur
Minerals
Micro elements- Manganese, iodine, copper,
cobalt, boron, molybdenum, iron and zinc
Energy source
Sucrose and glucose are preferred
and carbon
Thiamin, pyridoxine, nicotinic acid, biotin, citric
Vitamins
acid, ascorbic acid and inocitol etc.
Auxins, gibberellins, cytokinins, ethylene and
Growth regulators
abscisic acid etc.
Amino acids Glycine, tyrosine, L-cysteine and L-arginine etc.
Gelling agent Agar

Stock Solution
• Stock solution of various components are
prepared and used in the preparation of media.
• Sterilized by passing through bacteria proof filter
Amount of stock solution to be added (ml) =

Required concentration of media X Volume of media
______________________________________________________________________________________________________________________________________________________________
Concentration of stock solution X 1000

Preparation of Media
De-ionised DD Water in a flask (90% of total required volume)
Add the dehydrated medium into the water and stir to dissolve
the medium completely. Gently heat the solution.
Add desired heat stable supplements to the medium solution.
Add additional water to the medium solution to make up volume
Set the desired pH with dilute NaOH or HCl.
Sterilize the medium by autoclaving at 15 psi (121°C) for 15 min.
Add heat labile supplement (sterilized soln) after autoclaving.

Surface Sterilization of Seed to get
Aseptic Plant
Washing of seeds using 5% teepol soln and rinse with DD water
Rinse with 70% ethanol
Treat with 0.1% mercuric chloride (2-10 min.) / 10-12% hydrogen

peroxide (5-15 min.) / 1% silver nitrate (5-10 min.)
Rinse with DD water for two times.
Surface sterilized seed are germinated aseptically in a petridish, over a

cotton plug soaked with nutrient in continuous dark, at room temp.
Aseptic seedling → Aseptic plant
Collect explants from this aseptic plant.

Stem and aerial parts are sterilized by 70% ethanol and rinsed with DD water
Types of Culture
1. Callus Culture
Explants cultured on the appropriate medium (solid), with both
auxin and cytokinin
Give rise to an unorganised, growing and dividing mass of cells

(Callus tissue)
Sub-cultured on to fresh medium periodically
Callus culture is carried out in dark (due to lack of photosynthetic

process ) for 3-4 weeks.
Callus can also be used to initiate cell suspensions, which are used in a
variety of ways in plant transformation.
Manipulation of the auxin to cytokinin ratio in the medium can lead to
the development of shoots, roots or embryos from which,
whole plants can subsequently be produced.
The effect of different ratios of auxin to cytokinin on the growth and morphogenesis of callus
• Low auxin to cytokinin ratio (4:1) promote shoot development.

• High auxin to cytokinin ratio (100:1) promote root development.
• Intermediate ratio of auxin to cytokinin promote continued growth
of the callus without differentiation.
……Types of Culture
2. Cell-suspension Cultures
Callus is placed into a liquid medium in a ErlenMeyer flask.
Agitated (100-200 revolutions in a horizontal shaker)

Single cells are released into the medium.
Cultured for 4-6 weeks with frequent sub-culturing.

Released cells continue to grow and divide,
eventually producing
into the a cell-suspension culture
medium.
After subculture, the cells divide and the biomass of the culture
increases in a characteristic fashion until nutrients in the
medium are exhausted and/or toxic by-products build up to
inhibitory levels—this is called the ‘stationary phase’.
Growth Parameters and Growth
Curve
Total cell count
Fresh and dry weight of cells
Packed cell volume
Total protein and DNA content
Turbidity of the medium

Model growth showing different
growth phases in batch culture
3. Root Cultures
Root cultures can be established in-vitro on fairly simple media
from explants of
i. Root tip of either primary roots or lateral roots
ii. Root tip meristem of embryos
4. Shoot Tip and Meristem Culture

The tips of shoots (which contain the shoot apical meristem) can
be cultured in vitro, producing clumps of shoots.
5. Embryo Culture
Embryos can be used as explants to generate callus or somatic
embryos.
Both immature and mature embryos can be used as explants.
Embryo-derived embryogenic callus is the most popular
method of monocot plant regeneration.
6. Microspore Culture
Pollen contains the male gamete, which is termed as ‘microspore’.
Both callus and embryos can be produced from pollen.
Regeneration of plant from microspore explants can be

obtained by direct embryogenesis or via a callus stage
IMMOBILIZATION OF CELLS: It has been defined as a
technique,
• which confines the cells to a defined region in a space ,
• while retaining their catalytic activity,
• prevents its entry into the mobile phase, which carries the
substrate and product.
NEED FOR IMMOBILIZATION

• Protection from degradation and deactivation.
• Retention of enzyme, enzyme-free products.
• Cost efficiency.
• Enhanced stability.
 Use as controlled release agents.
 The ability to stop the reaction rapidly by removing the

enzyme from the reaction Solution (or vice-versa).
7. Immobilized Cell Cultures
Immobilized cells are obtained by encapsulating cell groups with a
suitable material like agarose or calcium alginate etc.
Immobilization of plant cells changes their cellular physiology in
comparison to suspension cultured cells.
These cells offers several advantages for their use in production of
phyto-chemicals at larger scale including single step bio-conversions.
Can be used for extended period of time.
Cells are packed in a column of a membrane and medium is allowed to
run through the column.
Slow growing cells accumulate larger metabolite than fast growing cells.
Loss of plant cells in suspension culture can be checked by immobilization.

8. Protoplast Culture
Protoplasts are plant cells with out cell wall.
Protoplasts are most commonly isolated from cell suspensions.
Removal of cell wall (usually by enzymatic hydrolysis, using cell

wall degrading enzymes like cellulase, hemicellulase and pectinase).
Protoplasts are fragile and easily get damaged.
Cultured carefully with out agitation in liquid medium.
High osmotic potential is maintained and aerated.
Genetic manipulation can be carried out in naked protoplast.

Genetic Transformation
Permanent incorporation of new or foreign DNA into genome of cell
Protoplast Fusion
Protoplasts from two different plants are mixed together and

forced to fuse.
Allowed for regeneration of cell wall.
Protoplasts can be plated out on to solid medium to get callus.
Regeneration by organogenesis or somatic embryogenesis from

callus.
Plant Regeneration
Whole plants can be regenerated from cultures in-vitro
Somatic embryogenesis
Organogenesis
→ Somatic embryo is
derived from a somatic cell.
→Embryo-like structures, →Formation of organs,
either directly from explants
which can develop into
whole plants. or from a callus culture.
A somatic cell is any cell of the body except sperm and egg cells. Somatic cells are diploid,
meaning that they contain two sets of chromosomes, one inherited from each parent.
A schematic representation of the sequential
stages of somatic embryo development
A simplified scheme for the integration of plant tissue culture
into plant transformation protocols.
Applications
1 Plant cells as bio-reactors: Production of the useful
natural compounds could be produced under controlled
environmental conditions, independent of soil and climate.
2 Study biogenesis of secondary metabolites using labeled

precursors.
3 Study the cytology and plant physiology
4 Genetic manipulation for better production of secondary

metabolites and disease resistant plants through protoplast
culture.
5 Production of disease-free plants: Plants grown from

aseptic plants
….Applications
6 Plant breeding
7 Micro-propagation: For large and continuous supply of plants
8 Cloning: Genetically identical plants derived from single explant

are called clones.
9 Tissue bank: Tissue are kept in frozen condition and cultured as

and when required; endangered species can be preserved.
10 Plant export and import: Aseptic plants, so easy to get

permission and requirement is less.
11 Immobilized plant cell culture in enhanced production of

phyto-chemicals.
……Applicat
ions
Bio-synthesis of medicinal compounds:
Plant cell culture technique is used for bio-transformation and
synthesis of those medicinal compounds which are too difficult or
impossible to synthesize chemically.
• Suspension cultures of a plant can be used for modification of a
substrate (bio-transformation).
✓ Diosgenin from Dioscorea, Sigmasterol from soyabean and
Deoxicholic acid from animal bile are used as starting material for
synthesizing steroidal structures by chemical reaction through many
steps. Example:
✓ Deoxicholic acid → Cortisone (Synthesis of cortisone require 31
steps by chemical synthesis)
……….Bio-synthesis of medicinal compounds
Digitalis lanata cells Hydroxylation reaction in production

medium containing precursors (-methyl
digitoxin to -methyl digoxin).
Podophyllum peltatum • Produce anti-cancer drug (etoposide) by

converting synthetic dibenzyl butanolides
→ lignans then Lignan → etoposide
(Conversion synthetically).
Rauwolfia serpentina Produces a new group of alkaloids namely
root cell culture raumacilines with high levels of ajmaline.
Cell suspension culture Morphine to codeine

of Ginkyo biloba
Applications in Production of
Phytoconstituents
• To date over 30 classes of therapeutically active compounds have
been produced in appreciable quantities in a bio-reactor., these
includes:
1. Digitalis glycosides
2. Rosmarinic acid
3. Opium alkaloids
4. Ginsenosides
5. Ajmacillin
6. Indole alkaloids like vinblastin and vincristin etc.
• Commercial production of a red pigment shikonin is carried
out by cell cultures.
Callus Culture of Optimization of growth
condition
secondary
Cell suspension
culture metabolite Process designing
producing plants
Enhancement of product
yield
Product
recovery
Purification
Specific product
[Procedure of process design and product recovery from the cultured plant cells ]
….Applications in Production of
Phytoconstituents
Bio-production of • Scopolamine, hyocyamin – Scopolia japonica.
metabolites in • Ajmacillin, serpentine and cantaratin – C. roseus.
hairy root culture
Bio-production of • Established for Belladona, Diascorea and Vinca.
metabolites in
shoot culture
Organogenesis • For production of the organ in which the specific
biochemical is formed.
Phytoconstituents Plant Increase in

phytoconstituents
Diasogenin D. deltadoea 7.8 %
Nicotine N. tobacum 5%
Serpentine C. roseus 2.2 %
Ajmacilline C. roseus 1.8 %
Anthraquinones Morinda citrifolia 18.8 %
General introduction, detailed study with respect to chemistry,
sources, preparation, evaluation, preservation, storage, therapeutic
used and commercial utility as pharmaceutical aids and/or
medicines for the following Primary metabolites:
1. Carbohydrates: Acacia, Agar, Tragacanth, Honey.
2. Proteins and Enzymes : Gelatin, casein, proteolytic enzymes

(Papain, bromelain, serratiopeptidase, urokinase, streptokinase,
pepsin).
3. Lipids(Waxes, fats, fixed oils) : Castor oil, Chaulmoogra oil, Wool

Fat, Bees Wax.
Dr. I. P. Padhy
Defination: “Carbohydrates are defined as polyhydroxy
aldehydes or polydroxy ketones or compounds on
hydrolysis produce either of the above.”
Carbohydrates are present universally in all plants and
animals.
CLASSIFICATION OF CARBOHYDRATES
Simple sugar (mono sacharides): Polysaccharides:
A. Bioses – Contain two carbon atoms A. Disaccharide: Composed of two mono
and do not occur free in nature. saccharide units (Sucrose = Glucose +
B. Trioses– Contain three carbon atoms fructose).
(glyceraldehydes) B. Tri Saccharides: (Raffinose = Glucose +
C. Tetroses - Contain four carbon Fructose + Galactose).
atoms ( erythrose) C. Oligosaccharides: Comprised of three to
D. Pentoses - Contain five carbon ten monosaccharide units e.g. Starchyose
atoms (ribose, xylose). (tetra sacharide).
E. Hexoses – Contain six carbon D. Poly saccharides: Indefinite number of
atoms: Glucose, fructose mono saccharide units (Starch, inuline,
cellulose etc.).
E. Gums: Guar gum
F. Mucilages: Isapgol husk
G. Pathological products
Gums and mucilage: Gums are either hydrophobic or hydrophilic high
molecular weight molecules, exhibit colloidal properties.
With appropriate solvent or swelling agent they produce gels, high viscose
suspensions or solutions.
Classification on the basis of their occurrence)
Seaweed Gum: Agar.

Natural 1. Plant exudates: Acacia, tragacanth
Gums 2. Seed gum: Guar Gum
3. Plant extract: Pectin.
Prepared 1.Biosynthetic gums: Xanthan

Gums 2.Starch and its derivatives
3.Cellulose derivatives: Carboxy methyl cellulose
Tests For Carbohydrates
• Molish test (general test): Substance + Molish reagent (α–
napthol and conc. H2SO4) → Purple colour.
• Tollens’ reagent test: Tollens reagent is a colorless, basic,
aqueous solution containing silver ions coordinated to
ammonia [Ag(NH3)2+]; give silver mirror in the inner wall of the
test tube when the sample contains aldose sugars.
• Selwinoff’s test (for keto-hexose like fructose): Heat 3 ml
Selwinoff’s regeant (resorcinol in concentrated HCl) and 1ml of
test solution in boiling water bath for 1-2 minutes → red colour
formed with keto sugars.
• Cobalt-Chloride test: Mix 3 ml test solution with 2 ml cobalt
chloride. Boil and cool. Add few drops NaOH Solution. The
solution appears greenish (in case of a aldose like glucose) or
purplish (in case of a ketose like fructose) and upper layer
greenish blue and lower layer purplish (in case of a mixture of a
aldose and a ketose for example glucose and fructose mixture).
Test for Reducing Sugar:
I. Reduction of felling’s solution: Substance (reducing sugar) + felling’s
solution. A and B heat → brick red precipitate
✓ Fehling's A is a blue aqueous solution of copper sulfate, while
Fehling's B is a clear and colorless solution of aqueous potassium
sodium tartrate.
II. Benedict’s reagent (aqueous solution of copper sulphate, sodium
carbonate and sodium citrate) + Test solution (reducing sugar) → Brick
red precipitate.
Test for non-reducing polysaccharides (starch):

I. Iodine test: Mix 3 ml of test solution and few drops of dilute iodine
solution. Blue colour appears which disappears on boiling and
reappears on cooling.
II. Tannic acid test for starch: With 20% tannic acid. Test solution
precipitate.
TEST FOR GUMS

a) Hydrolyze test solution using dilute Hcl. Perform Fehling’s and
Benedict’s test. Red colour precipitate is developed.
b) Swelling test: Powdered drug swells in water or aqueous KOH.
TEST FOR MUCILAGES

a) Powdered drug material shows pink/ red colour with ruthenium
red.
Agar (Agar-agar, Japanese-Isinglass, Vegetable gelatin)
Biological source: It is the dried colloidal concentrate from a

decoction of various red algae, particularly species of
• Gelidium: Gelidium amansii of family Gelidaceae
• Gracilaria of family Gracilaiaceae
Geographical source: Agar is produced commercially in Japan, New

Zealand, Australia, USA and India.
Collection and Preparation:
• In the coastal area of Japan, the algae are cultivated in special areas.
• The bamboo poles are planted (spreaded) in the sea to form
supports for the development of algae.
• The poles are withdrawn from time to time and the algae are
stripped off in the months from May to October.
• The algae are dried, beaten and shaken to remove any earthy
material adhering to it.
• It is then bleached by watering and drying in the sun.
• The algae are then boiled with acidulated water for several hours.
• A mucilaginous decoction is formed, which is filtered while hot
through a linen cloth.
• On cooling, a jelly is produced which is cut into bars and
subsequently strips are produced.
• The moisture is removed by freezing/ drying at about 35°C.
• The manufacturing of agar takes place only in winter season.
Description:
• Color: colorless to pale yellow
• Odor: not distinct odour
• Taste: mucilaginous
• Form: occurs in two forms: 1) Coarse powder or flakes
2) bundles of translucent crumpled strips
• Size: 2-5mm thick.
• Fracture: Tough when damp and brittle when dry.
• Solubility: In cold water does not dissolve but swells to a gelatinous
mass; in boiling water dissolves and 1% w/v solution gives a stiff jelly
on cooling.
Chemical Constituents:
It is a heterogeneous polysaccharide composed of two principal
constituents
→ Agarose (represents the gel strength )
→ Agaropectin (responsible for the viscosity of the agar solutions)
Chemical Tests:
1. Moisten the drug with a solution of Ruthenium red, a pink color is
produced due to mucilage.
2. Warm a small quantity of drug with caustic potash solution, a canary-
yellow color is produced.
3. Moisten the drug with N/50 iodine solution, a deep crimson color is
produced (different from Acacia and Tragacanth).
4. Heat a little drug in a test tube with soda-lime. Test the vapours with
litmus paper, no alkaline reaction (since no ammonia is produced).
5. Warm a little drug with acetic acid, formation of solution occurs on
prolonged heating.
Note: Tests 4 and 5 differentiate it from gelatin
Uses:
• Agar is used for the preparation of culture media
• It is used as an emulsifying agent
• It is a Bulk laxative and used in the treatment of constipation
• Used in affinity chromatography
Tragaranth (Gum tragacanth)
Biological source: it is the dried gummy exudation obtained by
incision from the steam and branches of Astragalus gummifer and
other species of Astragalus (family: Leguminosae).
Geographical source: Indigenous to Iran, Greece, Turkey, Iraq and

Syria. It is also found largely in India.
Collection and Processing:

• Palnt is a shrubs, thorny
• Gum excludes out immediately after an injury.
• Gum form as a result of transformation of the pith (soft and spongy
part present in the centre of stem) and medullary rays (A sheet of
vascular tissue separating the vascular bundles) into gummy substances.
• Incisions are done more on various parts of the stem and fluid
which oozes out is collected after drying (in April-November)
Macroscopy:
Colour – The flakes are white or
pale yellowish white.
Odour – Odourless.
Taste– Mucilaginous.
Shape– Thin, Flatted, ribbon like
flakes more or less curved.
Size – Flakes are approximately 25 x
12 x 2 mm.
Surface and Texture– Transverse
and longitudinal ridges are
present in the surface; texture is
rough.
Fracture – Short and horny
Solubility– Partly soluble in water,
in which it swells to homogenous
adhesive and gelatinous mass.
Chemical Constituents: It contains
• water soluble fraction (tragacathin)
• water insoluble fraction (bassorin)
Chemical Identification Tests:
• Tragacant Soln + few drops of aq. ferric chloride soln Boiled
deep yellow ppt.
• Sample solution + NaOH solution Canary yellow (light
warmed
to moderate yellow)
• Sample solution + strong iodine solution → green colour
Uses:
• Thickening, suspending and emulsifying agent
• Mucilage of tragacanth is used as binding agent, stabilizer in Ice
cream preparations.
• As demulcent (A medication in the form of an oily liquid or
semisolid that soothes inflamed or injured skin) in cosmetics.
Indian Gum: (Gum acacia, Gum arabic, Acacia)
Biological source: Acacia is the dried
gummy exudation from the stem
and branches of Acacia arabica,
Acacia senegal and of some other
species of Acacia of family
Leguminosae.
Geographical source: India, Srilanka,
Sudan (80% of total supply),
Morocco and Africa.
Cultivation and Collection: Evergreen
tree with short trunk. Gum is
collected from wild plants by
making incision.
Macroscopy:
Colour: Tears are nearly colourless or pale
amber; powder is light brown.
Odour: Odourless
Taste: Bland, mucilaginous.
Shape: Rounded, ovoid, or irregular
tears.
Size: Vary, usually about 0.5 to 6.0 cm. in
diameter
Textures: Glossy and marked with
minute fissures
Fracture: Brittle, breaking into
transparent, angular fragments with
glistening surfaces.
Solublity: Soluble in water, Insoluble in
95 % alcohol.
• Principally Arabin (Magnesium and calcium Salt of Arabic acid)
• Enzyme oxidase and Peroxidases
1. To 5 ml of a 2 per cent w/v test solution add 1 ml of strong lead
subacetate solution→a flocculent white precipitate is produced.
2. Dissolve 0.25 gm of sample in 5 ml. of water by shaking, add 0.5
ml of hydrogen peroxide solution and 0.5 ml of a 1% w/v
solution of benzidine in 90% alcohol, shake and allow to stand→
a deep blue colour is produced.
3. To 10 ml of a 2% w/v test solution, add 02 ml of a 20% w/v
solution of lead acetate→ no precipitate is produced.
4. To 0.1 g. of powder, add 1 ml. of N/ 50 iodine; the mixture does
not acquire a crimson or olive-green colour.
NOTE: Test no.- 3 and 4 is distinct from agar.
Uses:
• Administered intravenously in haemolysis.
• Used as demulcent, binding agent, suspending agent and

emulsifying agent.
• Along with Gelatin used in micro encapsulation process.
• Intravenously acacia has also been employed as a diuretic in

the treatment of nephrotic oedema.
HONEY
Synonyms: Madhu, Madh, Mel, Purified Honey.
Biological Source: Honey is a viscid and sweet secretion stored in

the honey comb by various species of bees, such as Apis mellifera,
Apis dorsata, Apis florea, Apis indica and other species of Apis,
belonging to family Apideae (Order: Hymenotera).
Geographical Source: Honey is available in abundance in Africa,

India, Jamaica, Australia, California, Chili, Great Britain and New
Zealand.
• The nectar of the flowers is a watery solution containing 25% sucrose and 75%
water.
• The worker bee sucks this nectar through its hollow tube of mouth (proboscis)
and deposits in honey-sac located in abdomen.
• The enzyme invertase present in saliva of the bee converts nectar into invert
sugar, which is partially utilized by the bee and the remaining is deposited into
honey comb.
• Honey comb is smoked to remove the bees and honey is obtained by applyng
the pressure to it or allowing it to drain naturally.
• The honey of commerce is heated to 80°C and allowed to stand.
• The impurities which float over the surface are skimmed off and the liquid is
diluted with water to produce honey of 1.35 density.
• Natural honey has the density of 1.47.
• Many-a-time, honey is extracted from the comb by centrifugation.
• It must be free from foreign substances.
• Honey is liable to fermentation, unless it is suitably processed.
• Honey is heated to 80°C before it is sent to the market, so as to avoid
fermentation.
• It should be cooled rapidly or else it darkens in colour on keeping.
• If necessary (and if not prepared by centrifugation method), honey is required
to be filtered through wet cloth or funnel.
Morphology
Chemical Constituents: The average composition of honey is as follows:

• Moisture 14–24%
• Dextrose 23–36%
• Levulose (Fructose) 30–47%
• Sucrose 0.4–6%
• Dextrin and Gums 0–7%
• Besides, it is found to contain small amounts of essential oil,
beeswax, pollen grains, formic acid, acetic acid, succinic acid,
maltose, dextrin, colouring pigments, vitamins and an admixture of
enzymes, for example, diastase, invertase and inulase.
Chemical Tests - Adulteration in honey is determined by the following tests:
1. Fiehe’s Test for Artificial Invert Sugar: Honey (10 ml) is shaken with
petroleum or solvent ether (5 ml) for 5–10 min. The upper ethereal layer
is separated and evaporated in a china dish. On addition of 1% solution
of resorcinol in hydrochloric acid (1 ml) a transient red colour is formed
in natural honey while in artificial honey the colour persists for
sometime.
2. Reduction of Fehling’s Solution: To an aqueous solution of honey (2 ml)

Fehling’s solutions A and B are added and the reaction mixture is heated
on a steam bath for 5–10 min. A brick red colour is produced due to the
presence of reducing sugars
Adulterant and Substitutes: Due to the relatively high price of pure honey,
it is invariably adulterated ether with artificial invert sugar or simply with
cane-sugar syrup. These adulterants or cheaper sub-stituents not only
alter the optical property of honey but also its natural aroma and
fragrance.
Uses
• Honey shows mild laxative, bactericidal, sedative, antiseptic and
alkaline characters.
• It is used for cold, cough, fever, sore eye
• Used in throat, tongue and duodenal ulcers, liver disorders,
scurvy and insomnia.
• It prevents infection and promotes healing.
• It is also useful in healing of carbuncles, chaps, scalds, whitlows
and skin inflammation.
• Used in the treatment of aphthae and other infection of the oral
mucous membrane.
• Honey is an important ingredient of certain lotions, cosmetics,
soaps, creams, balms, toilet waters and inhalations.
• Honey is used as an ingredient in various cough preparations.
• It is also used to induce sleep, cure diarrhoea.
24
“Lipids are the substances of animal or plant origin and comprise of fixed oils, fat
and waxes, chemically they are long chain fatty acids, alcohols or closely related
derivatives.”
Fixed oils, fats are glyceryl esters of higher Waxes are esters of fatty acids
long chain fatty acids. with high molecular weight
aliphatic monohydric alcohols.
Triglyceride: The major class of dietary lipids, including fats and oils
made up of 3 units of fatty acids and 1 unit called glycerol (backbone)
Glycerol Fatty Acids
is a • Unbranched carboxylic acids with 12-20 carbons.
trihydric • Melting points increase with increasing molecular weights.
alcohol • Unsaturation greatly lowers the melting point. 25
O
H2O O
R CH2 OH HO C R R CH2 O C R
+
Fatty alcohol Fatty acid Esterase (lipase) ester (lipid)
26
27
Common properties of fats and oils
• Greasy
• specific gravity is less than water and lighter than water.
• These are hydrophobic and lipophyllic in nature.
• Insoluble in water, sparingly soluble in alcohol and freely
soluble in solvents like petroleum ether, chloroform and
benzene.
• They leave a permanent translucent stain on white paper, so
called as fixed oils.
• They cannot be distilled, on heating, decompose and produce
an odour of scorched fat.
• Become rancid on long exposure to air (by oxidation), give
acidic reaction and disagreeable odour.
• Saponification process: Fats or waxes Hydrolysis with
alkali or enzyme → Free fatty acids + alkali → Salts (soaps)
28
Production of fixed oils and fats
Fixed oils and fats of vegetable origin are obtained by:
1. Extraction by expression: Fixed oils are obtained by expression in
hydraulic presses. If the expression is carried out in the cold, the oil is
known as a "virgin oil" or a "cold-pressed oil." In contrast, if the
expression is carried out in heat, the oil is known as a "hot-pressed
oil”.
2. Extraction by solvents: Sometimes organic solvents are used for
the extraction of oils.
• Animal fats are separated from other tissues by rendering with
steam, with or without pressure. The heat melts the fat, which
rises to the top and may be separated by decantation.
• After extraction these are refined by following various process
like degumming, neutralization, bleaching and de-orderisation
by injecting steam into very hot oil under vacuum.
29
Analytical parameters:
1. Acid value: Number of mg. of KOH required neutralizing the
free acids present in 1 gm of oil (high acid values indicate
rancified oils).
2. Saponification value: Number of mg. of KOH required to
neutralize the fatty acids resulting from complete hydrolysis of
1 gm of the oils.
3. Ester value: Ester value = Saponification value - Acid value.
4. Acetyl value: It is the number of milligrams of KOH needed to
neutralize the acetic acid liberated after hydrolysis of 1 gram of
acetylated fat (hydroxy fat first reacted with acetic anhydride).
5. Iodine value: It is the number of grams of iodine absorbed by
100 grams of fat or oil.
6. Physical parameters:
– Melting point for fats and waxes
– Specific gravity for oils
– Refractive index
– Viscosity
– Optical rotation
30
Tests for fats and oils
1. Filter paper gets permanently stained with oils.
2. Place a thick section of drugs on glass slide. Add a drop of Sudan
Red-III reagent. After 2 min. wash with 50% alcohol. Mount in
glycerin. Observe under microscope → oil globules appear red
3. To thin sections add a drop of 1% osmic acid, after one minute
observe under microscope → oil drops appear black
4. Extract + 2-3 drops of tincture alkane → gives red colour
5. Saponification test: 10 ml oil + 25 ml 10% NaoH Boil in boiling
water bath for 30 min. and cool + excess sodium sulphate solution
→ soap forms and rise to the top
6. Ethanolic solution of oil + few crystal of potassium hydrogen
sulphate Heat vigorously → pungent odour of acrylic aldehyde is
produced
7. Ethanolic extract + few drops of cupper sulphate solution + NaOH
solution → Clear blue solution is observed 31
Castor oil
Biological source: Castor oil is a vegetable oil obtained by
expression, from the seeds of
Ricinus communis (Euphorbiaceae).
Geographical source: India, Africa, Europe
Description: Castor Plant

• Colour: Colorless to very pale yellow,
very viscous liquid
• Odour: Mild or no odour
• Taste: Acrid
• soluble in ethanol
Castor seeds
32
Castor Oil Extraction
• Seeds are cleaned, cooked and dried prior to extraction
• Cooking is done to coagulate protein and to free the oil for efficient pressing.
• The first stage of oil extraction is pre-pressing, using a high pressure continuous
screw press – called the expeller.
• Extracted oil is filtered, and the material removed from the oil is fed back into the
stream along with fresh material.
• Material finally discharged from the press, called cake, contains 8 to 10 percent
oil. It is crushed and subjected to solvent extraction with hexane.
• Modification of the oil is achieved by a variety of chemical processes including
oxidation, hydrogenation and thermal treatments to produce products for specific
applications.
33
• It contains triglyceride in which approximately 90 % of ricinoleic
acid is present.
• Oleic, linoleic acids, iso ricinoleic acid, steric acid, and iso-steric
acid, are the other significant components.
• OIL must be free of ricin (toxic).
Chemical identification tests:

1. A mixture of 2 ml of the oil and 8 ml of ethanol (95%) is clear.
Uses:
• Laxative (A mild cathartic; stimulating evacuation of feces)
• Emollient (Having a softening or soothing effect especially to the
skin); used in the preparation of lipsticks
• Used in the preparation of hair creams, hair fixtures.
• Substitute of Spermaceti, bees wax, carnauba wax, in the
preparation of ointments and creams.
34
Bees wax (Yellow bees wax, Cera-flava)
Biological source: It is the purified wax obtained from the honey
comb of the bees Apis mellifera, Apis dorsata and other species of
Apis of family: Apidae
Geographical source: It is processed and commercially prepared in
France, Italy, West-Africa, Jamaica and India.
Preparation:
• Honey comb are broken and boiled in water by keeping in porous
bags.
• Boiling causes oozing of wax which gets collected out side the bag
and forms a cake after cooling.
• Purified by heating in boiling water or dilute sulfuric acid followed
by settling, then are skimmed off.
• Bleached using hydrogen per oxide/ ozone/ chromic acid/ charcoal
or chlorine.
35
Description:
• Colour: Yellow to yellowish brown, non crystalline solid.
• Odour: Agreeable and honey like.
• Texture: Soft to touch.
• Solubility: Insoluble in water, soluble in hot alcohol, chloroform,
CCl4, fixed oils and volatile oils.
Chemical constituants:
• It contains esters of straight chain monohydric alcohols with
straight chain acids.
• The chief constituents are myricine, free cerotic acid.
• Aromatic substance cerolein is also present in the wax.
Uses:
• Used in the preparation of ointments, plasters, polishes, lip-
sticks and face creams.
• It is an ingredient of paraffin ointment
36
Lanolin/ WOOL FAT
Synonyms: Oesipos; Agnin; Alapurin; Anhydrous lanolin; Adeps
lanae; Laniol.
Biological Source: Lanolin is the fat-like purified secretion of the
sebaceous glands which is deposited into the wool fibres of
sheep, Ovis aries Linn., belonging to family Bovidae.
Preparation:
• Wool is cut and washed with a soap or alkali.
• An emulsion of wool fat, called as wool grease, takes place in
water.
• Raw lanolin is separated by cracking the emulsion with sulphuric
acid.
• Wool grease floats on the upper layer and fatty acids are
dissolved in the lower layer.
• Lanolin is purified by treating with sodium peroxide and bleaching
with reagents.
Characteristics
• Lanolin is a tenacious, unctuous mass.
• Yellowish white
• Odour is slight and characteristic.
• Practically, it is insoluble in water, but soluble in chloroform or
ether with the separation of the water.
• It melts in between 34 and 40°C.
• On heating it forms two layers in the beginning, continuous
heating removes water.
• Lanolin is not saponified by an aqueous alkali. However,
saponification takes place with alcoholic solution of alkali.
• Anhydrous lanolin is a yellowish tenacious, semisolid fat with
slight odour. Practically it is insoluble in water but mixes with
about twice its weight of water without separation. It is freely
soluble in benzene, chloroform, ether, carbon disulphide, acetone,
and petroleum ether.
• Lanolin is a complex mixture of esters and polyesters of 33 high molecular
weight alcohols, and 36 fatty acids.
• The chief constituents of lanolin are cholesterol, iso-cholesterol,
unsaturated monohydric alcohols of the formula C27H45OH, both free and
combined with lanoceric, lanopalmitic, carnaubic, and other fatty acids.
• Lanolin also contains esters of oleic and myristic acids, aliphatic alcohols,
such as cetyl, ceryl and carnaubyl alcohols, lanosterol, and agnosterol.
Identification Tests: Dissolve 0.5 g of lanolin in chloroform, and to it add 1
ml of acetic anhydride and two drops of sulphuric acid. A deep green
colour is produced, indicating the presence of cholesterol.
Uses
• Lanolin is used as an emollient, as water absorbable ointment base in
many skin creams and cosmetic and for hoof dressing.
• Wool fat is readily absorbed through skin and helps in increasing the
absorption of active ingredients incorporated in the ointment.
• However, it may act as an allergenic contactant in hypersensitive persons.
CHAULMOOGRA OIL
Synonyms: Hydnocarpus oil; gynocardia oil.
Biological Source: Chaulmoogra oil is the fixed oil obtained by cold

expression from ripe seeds of Hydnocarpus anthelminticta, Hydnocarpus
heterophylla, and other species of Hydnocarpus, belonging to family
Flacourtiaceae.
Geographical Source: The plants are tall trees, up to 17 m high, with narrow
crown of hanging branches; native to Burma, Thailand, eastern India, and
Indo-China.
Characteristics:
• The oil is yellow or brownish yellow.
• Below 25°C it is a soft solid.
• It has peculiar odour and sharp taste.
• It is soluble in benzene, chloroform, ether, petrol; slightly soluble in cold
alcohol; almost entirely soluble in hot alcohol and carbon disulphide.
• Chaulmoogra oil contains glycerides of cyclopentenyl fatty acids like
hydnocarpic acid (48%), chaulmoogric acid (27%), gorlic acid with small
amounts of glycerides of palmitic acid (6%), and oleic acid (12%).
• The cyclic acids are formed during last 3–4 months of maturation of the
fruit and are strongly bactericidal towards the Micrococcus of leprosy.
Uses
• The oil is useful in leprosy and many other skin diseases.
• The cyclopentenyl fatty acids of the oil exhibit specific toxicity for
Mycobaeterium leprae and M. tuberculosis.
• The oil has now been replaced by the ethyl esters and salts of
hydnocarpic and chlumoogric acids.
• At present organic sulphones have replaced Chaulmoogra oil in
therapeutic use.
Proteins and Enzymes :
Gelatin, Casein and Proteolytic Enzymes (Papain,
bromelain, serratiopeptidase, urokinase,
streptokinase, pepsin).
GELATIN
Synonyms: Gelfoam; puragel; gelatinum.
Biological Source: Gelatin is a protein derivative obtained by evaporating an

aqueous extract made from bones, skins, and tendons of various domestic
animals. Some important sources are: Ox, Bos taurus, and Sheep, Ovis aries
belonging to family Bovidae
Characteristics
• Gelatin occurs as a transparent, brittle, sheet, flakes or course granular powder
• Colourless or slightly yellow, Odourless, Tasteless.
• In water it swells and absorbs 5–10 times its weight of water to form a gel in
solutions below 35–40°C.
• It is insoluble in cold water and organic solvents, soluble in hot water, glycerol,
acetic acid; and is amphoteric.
• In dry condition it is stable in air, but when moist or in solution, it is attacked by
bacteria.
• The gelatinizing property of Gelatin is reduced by boiling for long time.
• The quality of gelatin is determined on the basis of its jelly strength (Bloom
strength).
• Jelly strength is used in the preparation of suppositories and pessaries.
Preparation: The process of manufacture of gelatin vary from factory to
factory. However, the general outline of the process is given below.
• Raw material: Bones, skins, and tendons of Bovideans is collected and subjected
to liming operation.
• Liming Process: The raw material is first subjected to the treatment known as
‘liming’. In this process, the skins and tendons are steeped for fifteen to twenty
and sometimes for 40 days in a dilute milk of lime. During this, fleshy matter gets
dissolved, chondroproteins of connective tissues gets removed and fatty matter is
saponified. The animal skin is further thoroughly washed in running water.
• Defattying: In case of bones, the material is properly ground and defatted in close
iron cylinders by treatment with organic solvents such as benzene. The mineral
and inorganic part of the bone is removed by treatment with hydrochloric acid.
• Extraction: The treated material from bones, skins and tendons is boiled with
water in open pans with perforated false bottom. This process can also be carried
out under reduced pressure. The clear liquid runs of again and again and is
evaporated until it reaches to above 45 per cent gelatin content.
• Setting: The concentrated gelatin extract is transferred to shallow metal trays or
trays with glass bottom. It is allowed to set as a semisolid jelly.
• Drying: The jelly is transferred to trays with a perforated wire netting bottom and
passed through series of drying compartments of 30–60°C increasing each time
with 10°C. About a month is taken for complete drying.
• Bleaching: In case of darker colour, finished product is subjected to bleaching by
sulphur dioxide. Bleaching affords a light coloured gelatin.
• Gelatin consists of the protein glutin which on hydrolysis gives a
mixture of amino acids.
• The approximate amino-acid contents are: glycine (25.5%), alanine
(8.7%), valine (2.5%), leucine (3.2%), isoleucine (1.4%), cystine and
cysteine (0.1%), methionine (1.0%), tyrosine (0.5%), aspartic acid
(6.6%), glutamic acid (11.4%), arginine (8.1%), lysine (4.1%), and
histidine (0.8%).
• Nutritionally, gelatin is an incomplete protein lacking tryptophan.
• The gelatinizing compound is known as chondrin and the adhesive
nature of gelatin is due to the presence of glutin.
Chemical Tests:
1. Biuret reaction: To alkaline solution of a protein (2 ml), a dilute solution
of copper sulphate is added. A red or violet colour is formed with
peptides containing at least two peptide linkages. A dipeptide does not
give this test.
2. Xanthoproteic reaction: Proteins usually form a yellow colour when
warmed with concentrated nitric acid. This colour becomes orange when
the solution is made alkaline.
3. Millon’s reaction: Millon’s reagent (mercuric nitrate in nitric acid
containing a trace of nitrous acid) usually yields a white precipitate on
addition to a protein solution which turns red on heating.
4. Ninhydrin test: To an aqueous solution of a protein an alcoholic solution
of ninhydrin is added and then heated. Red to violet colour is formed.
5. On heating gelatin (1 g) with soda lime, smell of ammonia is produced.
6. A solution of gelatin (0.5 g) in water (10 ml) is precipitated to white buff
coloured precipitate on addition of few drops of tannic acid (10%).
7. With picric acid gelatin forms yellow precipitate.
Uses
• Gelatin is used to prepare pastilles, pastes, suppositories,
capsules cells, pill-coatings, gelatin sponge; as suspending
agent, tablet binder, coating agent, as stabilizer, thickener and
texturizer in food.
• It forms glycerinated gelatin with glycerin which is used as
vehicle and for manufacture of suppositories.
• Combined with zinc, it forms zinc gelatin which is employed as
a topical protectant.
• As a nutrient, Gelatin is used as commercial food products and
bacteriologic culture media.
• It is also used for manufacturing rubber substitutes, adhesives,
cements, lithographic and printing inks, plastic compounds,
artificial silk, photographic plates and films, light filters for
mercury lamps, clarifying agent, in hectographic matters, sizing
paper and textiles, for inhibiting crystallization in bacteriology,
for preparing cultures and as a nutrient.
CASEIN
Biological Source:
• Casein is a proteolytic enzyme obtained from the stomachs of
calves.
• It is extracted from the proteins of the milk; in the milk.
• The casein content of milk represents about 80% of milk proteins.
Characteristics:
• The isoelectric point of casein is 4.6.
• The purified protein is water insoluble.
• While it is also insoluble in neutral salt solutions, it is readily
dispersible in dilute alkalis and in salt solutions such as sodium
oxalate and sodium acetate.
• Casein does not coagulate on heating.
• It is precipitated by acids and by a proteolytic enzyme (rennet).
• Milk consists of 80% of milk proteins (casein).
• The major constituents of casein are alpha (s1) and alpha
(s2)-caseins, β-casein and kappa-casein.
• These caseins are conjugated proteins with phosphate
group(s) which are esterified into serine residues they
have a low solubility at pH 4.6.
Uses:
• It is used in the manufacture of binders, adhesives,
protective coatings and food additives.
• It is commonly used by bodybuilders as a slow-digesting
source of amino acids.
• There is growing evidence that casein may be addictive for
some individuals, particularly those on the autism
spectrum or having schizophrenia.
Enzymes
• Organic catalyst produced by the body by living organisms.
• They perform many complex chemical reactions that make up
life processes.
• They are lifeless, and when isolated, they still exert their
characteristic catalytic property.
• They are colloids, soluble in water and dilute alcohol
• Precipitated by concentrated alcohol
• Enzymes are sensitive to heat and are denatured by excess
heat or cold
• Most enzymes are best at temperature between 35-40°C.
• Above 65°C with the presence of moisture they get destroyed.
• Their activity is negligible at 0°C.
• Enzymes are sensitive to pH.
• Enzymes are created in cells but are capable of functioning out
side of the cell.
• Enzymes are reusable
Further on the basis of site of action, enzymes can
be classified under two categories:
(a) Endoenzymes (Intracellular enzymes):
Enzymes which act only inside of the cell are
known as endoenzymes, e.g. digipuridase,
phosphorylases.
(b) Exoenzymes (Extracellular enzymes):
Enzymes which act or are active outside the cell
are known as enxoenzymes, e.g. all digestive
enzymes [amylases].
Proteolytlc Enzymes:
There are mainly three proteolytic enzymes namely:
(A) Papain: Enzyme obtained from plant.
(B) Pepsin: Both available in humans and other animals.
(C) Trypsin: Found in the digestive system of many
vertebrates.
PAPAIN
SYNONYM: Papayotin , vegetable pepsin, tromasin
BIOLOGICAL SOURCE: Dried purifed latex of the green fruits and

leaves of the Carica papaya of family Caricaceae.
GEOGRAPHICAL SOURCE: America, Sri Lanka, Florida, India, Cuba.
PREPARATION:
• Longitudinal scratches are made in the skin of the immature
fruit while it is still hanging on the tree.
• Incisions and collection are made at weakly intervals.
• Fruit exudes the latex (between 5 and 10 A.M.).
• The lumps are shredded and dried in sun or by the means of
artificial heat.
• It is purified by dissolving in water and precipitating with
alcohol.
PROPERTIES:
Colour: Light brown to white coloured amorphous
powder.
Odour: Typical odour
Taste: Typical taste
Solubility: It is soluble in water.
Other Characters:
• It has maximum activity at pH 5 to 6
• It is much less stable than pepsin and trypsin,
particularly in the presence of oxygen.
CHEMICAL CONSTITUENTS: It contains several enzymes that
include one or more proteolytic enzymes:
1. Papain, a coagulating enzyme which acts upon the casein of milk;
2. an amylolytic enzyme
3. a clotting enzyme similar to peptase
• It is quite apparent that more than one proteolytic enzyme is
present because a single sample of papain will yield variable
results depending upon the protein used in the substrate.
• The best grade digests 300 times it’s weight of egg albumin.
CHEMICAL TEST
USES:
• Papain is used as a digestant for proteins
• as an ingredient in cleaning solutions for soft
contact lenses.
• Papain is used extensively for tenderizing beef.
• It is used in meat packing industries.
• It is used in relieving symptoms of episiotomy
(An episiotomy also known as perineotomy, is a surgically planned incision on
the perineum and the posterior vaginal wall during second stage of labor).
PEPSIN
BIOLOGICAL SOURCE: It is a proteolytic enzyme obtained
from the glandular layer of the fresh stomach of the hog
Var domesticus ; Family: Suidae.
PREPARATION:
• Mucous membrane is separated from the stomach by the
process of stripping or scrapping.
• Placed in the acidified water for autolysis at 37°C for 2 hrs.
• The liquid obtain contains pepsin and peptone.
• Filter it
• Add sodium or ammonium salts to the filtrate, until it is
half saturated.
• At this point, pepsin separates out while peptone remains
in the solution.
• Pepsin is collected and dried at low temperature.
DESCRIPTION:
Colour: Pale yellow coloured translucent grains
Odour: Very faint odor
Taste: Slightly bitter taste
Solubility: Soluble in water, acids and NaCl Solution.
Other characters:
✓ Best active at 40°C with 2-4 pH.
✓ Unstable above 6 pH
✓ Denature above 70°C.
✓ It can be stored for 2 years in 2-8°C
CHEMICAL TEST: Same as papain but only difference is pH, which has to be
adjusted to 2 for the test. It is done by addition of HCl.
USES:
• Supplemented in the deficiency of gastric secretion.
• Used in the various analysis of proteins in the laboratory.
• In the preparation of cheese and other protein containing foods.
SERRATIOPEPTIDASE
Synonym: Serrapeptase, serratiopeptidase.
Biological Source:
• Serratiopeptidase is a proteolytic enzyme isolated from nonpathogenic
Enterobacteria Serratia E 15 (produced by fermentation technology by
using nonpathogenic enterobacteria species such as Serratia E 15.).
• It is also produced by the larval form of the silk moth (The larvae of silk
moth produce this enzyme in their intestine to break down cocoon walls.
It can thus be obtained from the silk moth larvae).
Characteristics:
• Serratiopeptidase is very much vulnerable to degradation in the acidic pH.
• When consumed in unprotected tablet or capsule, it is destroyed by acid
in stomach.
• However enteric coated tablets facilitate its absorption through intestine.
• One unit of the enzyme hydrolyses casein to produce colour equivalent to
1.0 μmol of tyrosine per minute at pH 7.5 and 35°C.
Chemical Constituents: Serratiopeptidase is a proteolytic enzyme
of protease type. The preparation contains 7.1 units/mg solid.
Uses
• Serratiopeptidase is the most widely prescribed
antiinflammatory enzyme in developed countries and also in
India.
• It eliminates inflammatory oedema and swelling, accelerate
liquefaction of pus and sputum, and enhance the action of
antibodies.
• It is also used as a fast wound healing agent.
• It is proving to be a superior alternative to the nonsteroidal
antiinflammatory drugs traditionally used to treat rheumatoid
arthritis and osteoarthritis. I
• t has wide ranging applications in trauma surgery, plastic
surgery, respiratory medicine, obstetric and gynaecology.
UROKINASE
Synonym: Uroquinase.
Biological Source: Urokinase is serine protease enzyme isolated from
human urine and from human kidney cells by tissue culture or by
recombinant DNA technology.
Preparation:
• Urokinase is a fibrinolytic enzyme produced by recombinant DNA
using genetically manipulated E. coli cells.
• It is produced firstly as prourokinase and then converted to active
form by plasmin or kallikrein.
• Urokinase used medicinally is also purified directly from human urine.
• It binds to a range of adsorbents such as silica gel or kaolin which can
be use to initially concentrate and purify the product.
• It can be further purified by precipitation with sodium chloride or
ethanol or by chromatography.
• Human urokinase needs sterile filtration, a septic filling and freeze
drying.
Characteristics:
• Urokinase enzyme occurs in two different forms as single and double
polypeptide chain forms.
• It has a half-life of 10–16 minutes after intravenous administration.
• These enzymes act on an endogenous fibrinolytic system.
• Urokinase enzymes are serine proteases that occur as a single low
molecular weight (33 kDa) and double, high molecular weight (54 kDa)
• They differ in molecular weight considerably.
• A single chain is produced by recombinant DNA technique and is known
as SCUPA.
Uses:
• Urokinase is used in the treatment of pulmonary embolism, coronary
artery thrombosis and for restoring the potency of intravenous
catheters.
• It is generally administered intra-venously in a dose of 4,400 units/kg
body weight per hour for twelve hours.
STREPTOKINASE
Synonym: Estreptokinase, plasminokinase.
Biological Source: Streptokinase, is a sterile, purified preparation of a

bacterial protein elaborated by group C (beta) -hemolytic streptococci-
S. griseus.
Preparation:
• Streptokinase is a bacterial derived enzyme of serine protease group.
• Streptokinase is produced by fermentation using streptococcal culture
and is isolated from the culture filtrate.
• It is produced in the form of a lyophilized powder in sterile vials
containing 2,50,000 to 7,50,000 IUs.
Characteristics:
• Streptokinase is a bacterial protein with half-life of 23 minutes.
• Its anisolylated plasminogen activator complex (APSAC) has a higher
half-life of six hours.
Chemical Constituents: Streptokinase is the purified bacterial protein with
about 484 amino-acid residues.
Uses:
• Streptokinase is the first available agent for dissolving blood clots.
• It binds to plasminogen in a 1:1 ratio and changes molecular
conformation.
• Thus, the complex formed becomes an active enzyme and promotes
the activity of fibrinolytic enzyme plasmin.
• Plasmin breaks fibrin clots.
• Anistreptase or the anisolylated plasminogen streptokinase activator
complex (APSAC) can also be used in a similar way for degrading blood
clots.
• Streptokinase and anistreptase are both used in the treatment of
pulmonary embolism, venous and arterial thrombosis and coronary
artery thrombosis.
• It is also sometimes administered along with heparin to counter act a
paradoxical increase in local thrombin.
BROMELAIN
Synonyms: Bromelin, bromelain.
Biological Source: Bromelin is a mixture of proteolytic enzymes
isolated from the juice of Ananas comosus (pineapple), belonging
to family Bromeliaceae.
Geographical Source: Pineapple is a native of tropical America. It is
grown in almost all parts of the world including India, China, Thai-
land, United States, Brazil, Philippines, Mexico, Hawaii, and
Taiwan.
Characteristics:
• It is obtained in light brown-coloured powder.
• The optimum pH of bromelain is 5.0–8.0.
• In solution pH below 3.0 and above 9.5 inactivates the enzyme.
• The optimum temperature is between 50 and 60°C, still it is
effective between 20 and 65°C too.
• The moisture content should not exceed 6%.
Cultivation, Collection, and Preparation
• Bromelin is found in pineapple fruit juice and stem.
• Pine-apple is perennial, and it does not have a natural period of
dormancy.
• It is propagated through suckers, slips, and crowns.
• In India it is planted in August, the plant generally flowers in
February–March, and the fruit ripens during July–October.
• The fruits must be left on the plant to ripen for the full flavour to
develop.
• Dark green unripe fruits gradually change to yellow and finally to
deep orange. The fruits are cut off.
• The enzyme bromelin does not disappear as the fruit ripens.
• The enzyme from fruit and stem are known as fruit bromelin and
stem bromelin, respectively.
• It is isolated from pineapple juice by precipitation with acetone
and also with ammonium sulphide.
• Bromelain is not a single substance, but
rather a collection of enzymes and other
compounds.
• It is a mixture of sulphur-containing
protein-digesting enzymes, called
proteolytic enzymes or proteases.
• It also contains several other substances in
smaller quantities, including peroxidase,
acid phosphatase, protease inhibitors, and
calcium.
Uses
• Bromelain is an effective fibrinolytic agent.
• It inhibits platelet aggregation and seems to have both direct as well as
indirect actions involving other enzyme systems.
• It can modify the permeability of organs and tissues to different drugs.
• The potentiation of antibiotics and other medicines by bromelain may be
due to enhanced absorption, as well as increased permeability of the
diseased tissue which enhances the access of the antibiotic to the site of
the infection.
• It is also thought that the use of bromelain may provide a similar access to
specific and nonspecific components of the immune system, therefore,
enhancing the body’s utilization of its own healing resources.
• Bromelain has been used successfully as a digestive enzyme following
pancreatectomy, in cases of exocrine pancreas insufficiency and in other
intestinal disorders.
• Research has indicated that bromelain prevents or minimizes the severity
of angina pectoris and transcient ischemic attacks (TIA).
• It is useful in the prevention and treatment of thrombosis and
thrombophlebitis.
• It may even be useful in the treatment of AIDS to stop the spread of HIV.
• It has no major side effects, except for possible allergic reactions.
• Conservation is the process of management of
biosphere in order to obtain the greatest benefit for
the present generation and maintaining the potential
for future.
• Conservation of medicinal plant resources is of global
concern because we don't know what we are losing
and what we will need in future.
Dr. I. P. Padhy
means plant.


quantities.
Properties
• Odourless.
solvents.
parts of water.
(Tubarcuranine Cl.)
processes
Types of alkaloids
Homoatropine.
Source
lysine).
acid pathway.
Classification
Classification
General tests:
Specific tests:
observed.
is produced.
colour.
GLYCOSIDES

Hydrolysis

Properties:
General Uses:
expectorant etc.
Classification






smoky flame
Classification
VOLATILE OILS
or aetherolea.
Properties
soluble in alcohol.
Source:
trichomes etc.

Classification
fennel.


Classification
Dr. I. P. Padhy
INTRODUCTION
the world.
agents.
CLASSIFICATION:
2.Marine bacterium
Active Source Use
Compounds
angustata.
giganteus.
ANTICANCER DRUGS
Active Source Use
Compounds
and spongouridine.
gorganian.
angasi.
antifungal.
zonaroids)
hemifera)
Aeroplysinin-1(-)
mammosa)
ANTIBIOTICS

acremonium)
phenol.
MISCELLANEOUS
sponge Tethya crpta
(antispasmodics)
halibut liver oil
variabilis)
Dr. I. P. Padhy
INTRODUCTION
the world.
agents.
CLASSIFICATION:
2.Marine bacterium
Active Source Use
Compounds
angustata.
giganteus.
ANTICANCER DRUGS
Active Source Use
Compounds
and spongouridine.
gorganian.
angasi.
antifungal.
zonaroids)
hemifera)
Aeroplysinin-1(-)
mammosa)
ANTIBIOTICS

acremonium)
phenol.
MISCELLANEOUS
sponge Tethya crpta
(antispasmodics)
halibut liver oil
variabilis)
Prof. I. P. Padhy
Tissue culture
Concept
from a single cell.
1896
differentiate
Totipotency
History
1920
root of tomato



plant cells
Stages

1

Functions
Water
Growth regulators

Organic compounds


Minerals
Energy source
and carbon
Vitamins
Growth regulators
abscisic acid etc.
Gelling agent Agar

Stock Solution

______________________________________________________________________________________________________________________________________________________________


Aseptic Plant



Types of Culture
1. Callus Culture
auxin and cytokinin

(Callus tissue)




medium.
Curve
Total cell count
Packed cell volume

3. Root Cultures
from explants of

5. Embryo Culture
embryos.

technique,





Protoplast Fusion

forced to fuse.

callus.
Plant Regeneration
Organogenesis
Applications

precursors.

culture.

aseptic plants
….Applications
6 Plant breeding

are called clones.



phyto-chemicals.
……Applicat
ions
steps. Example:



of Ginkyo biloba
Phytoconstituents
includes:
2. Rosmarinic acid
3. Opium alkaloids
4. Ginsenosides
5. Ajmacillin
condition
secondary
Cell suspension
producing plants
yield
Product
recovery
Purification
Specific product
Phytoconstituents
hairy root culture
metabolites in
shoot culture

phytoconstituents

pepsin).

Fat, Bees Wax.
Dr. I. P. Padhy
animals.
cellulose etc.).
E. Gums: Guar gum
Seaweed Gum: Agar.


sodium tartrate.
red precipitate.

precipitate.
TEST FOR GUMS

TEST FOR MUCILAGES

red.


Description:
on cooling.
constituents
Chemical Tests:
prolonged heating.
Uses:


Macroscopy:
12 x 2 mm.
rough.
deep yellow ppt.
warmed
to moderate yellow)
Uses:
cream preparations.
Leguminosae.
Morocco and Africa.
making incision.
Macroscopy:
Odour: Odourless
tears.
diameter
minute fissures
95 % alcohol.
Uses:

emulsifying agent.

HONEY


Zealand.
water.
honey comb.
fermentation.
Morphology

sometime.

fragrance.
Uses
mucous membrane.
24
derivatives.”
O
H2O O
+
26
27
• Greasy
benzene.
28
oil”.
29
rancified oils).
1 gm of the oils.
– Viscosity
30
produced
Castor oil

very viscous liquid
• Taste: Acrid
Castor seeds
32
applications.
33
acid is present.

Uses:
34
Preparation:
bags.
or chlorine.
35
Description:
Uses:
36
Lanolin/ WOOL FAT
lanae; Laniol.
Preparation:
water.
acid.
with reagents.
Characteristics
• Yellowish white
Uses
CHAULMOOGRA OIL

Flacourtiaceae.
Indo-China.
Characteristics:
Uses
therapeutic use.
GELATIN

Characteristics
bacteria.
strength).
histidine (0.8%).
Chemical Tests:
give this test.
Uses
texturizer in food.
CASEIN
Biological Source:
calves.
Characteristics:
Uses:
Enzymes
life processes.
heat or cold
side of the cell.
phosphorylases.
enzymes [amylases].
vertebrates.
PAPAIN

PREPARATION:
artificial heat.
alcohol.
PROPERTIES:
powder.
Other Characters:
CHEMICAL TEST
USES:
contact lenses.
PEPSIN
PREPARATION:
• Filter it
half saturated.
in the solution.
DESCRIPTION:
Other characters:
USES:
SERRATIOPEPTIDASE
Biological Source:
Characteristics:
in stomach.
Uses
India.
antibodies.
UROKINASE
Preparation:
drying.
Characteristics:
as SCUPA.
Uses:
catheters.
STREPTOKINASE

S. griseus.
Preparation:
Characteristics:
Uses:
conformation.
clots.
artery thrombosis.
BROMELAIN
Taiwan.
Characteristics:
dormancy.
develop.
compounds.
calcium.
Uses
the infection.
thrombophlebitis.
PHARMACOGNOSY IN VARIOUS SYSTEMS OF MEDICINE:
ROLE OF PHARMACOGNOSY IN ALLOPATHY AND

TRADITIONAL SYSTEMS OF MEDICINE NAMELY, AYURVEDA,
SIDDHA, UNANI, HOMEOPATHY AND CHINESE SYSTEMS OF
MEDICINE.
Dr. I. P. Padhy
Traditional Medicine (according to WHO): “The health practices
which approaches knowledge and beliefs; incorporating plant,
animal and mineral-based medicines, spiritual therapies, manual
techniques and exercises, applied singularly or in combination, to
treat, diagnose and prevent illnesses or maintain well-being.
Based on past experience and observations, handed down from

generation to generation verbally or in writing within various
societies that developed before the era of modern medicine.
2
Practices known as traditional medicines include:
• Ayurveda
• Siddha medicine
• Unani (Greco-Arabic medicine)
• Ancient Iranian medicine
• Islamic medicine
• Traditional Chinese medicine
• Chinese herbal medicine
• Acupuncture and moxibustion
• Acupressure
• Traditional Korean medicine
• Traditional Vietnamese medicine
• Traditional African medicine
3
Terminologies
Alternative system of Medicine:
It is any practice claiming to heal, that does not fall within the domain
of conventional medicine.
Complementary Medicine:
Any alternative system, used together (complementary) with
modern medicine or other therapies.
Example: Homoeopathy, aromatherapy, acupressure and
acupuncture, yoga etc.
The terms "complementary medicine“ and "alternative medicine"

are used inter-changeably with traditional medicine. 4
Terminologies
Folk Medicine: It refers to the healing practices and health

preservation known to a limited segment of the population in a
culture, transmitted informally as general knowledge and practiced
or applied by any one in the culture having prior experience.
Native Medicine: It is a derogatory version of traditional medicine.
The term native meaning anything not foreign or not introduced by
colonial masters.
5
Terminologies
Medicinal Plant: Any plant or plant part contains substances
which change beneficially the physiology, that can be used for
therapeutic purposes or which are precursors for the synthesis of
useful drugs.
Herb: In botany, is a plant that does not form a woody stem.
Medicinal Herb: It is different from botanic term “herb”.

It refers to any plants used for medicinal purposes. For example, a
medicinal herb can be a real herbal plant, a shrub, other woody
plant or a fungus. 6
Terminologies
Sacred Herbs: Herbs are used in many religions for various religious
purposes; example: Myrrh (Commiphora myrrha) in Christianity and
holy basil or Tulsi (Ocimum species) in Hinduism etc.
Culinary Herbs: These are vegetable drugs which provide aroma.
These are used for both as spices and medicinal purposes.
Vegetable Drugs: Medicinal plants or their parts like leaf, bark, root
etc., having cellular parts, used for therapeutic purposes.
7
Expressed Juice: Liquids or saps squeezed from plant material and
either applied orally or externally.
Poultice: Expressed juice applied to a inflammation, wound or sore

either directly or wrapped in cloth, sometimes after heating.
Infusion, Tisane or Tea: This is a preparation of medicinal herbs for
internal or external application prepared after steeping a small quantity
of herb with hot water.
Decoction: This is a extract prepared by boiling the powdered herb.
[about 28 to 56 grams (1 to 2 ounces) of herb to 0.5 liter of water].
Concoction: An minced, dissolved, or macerated preparation from
combination of various ingredients, usually herbs, spices, condiments or
minerals. 8
Ethno-botany is the study of the relationship between plants,
people and their culture.
Ethno-pharmacology is the scientific study of ethnic groups and
their use of drugs (plant, animal or mineral).
Ethno-pharmacy is the interdisciplinary science that investigates
the perception and use of pharmaceuticals (especially in
traditional medicines) with in a given human society.
9
Traditional Medicine vs. Modern Medicine.
• Object is same i.e. to cure the diseases or preventing the diseases.
• Differs in the concept, etiology, method of diagnosis and approach
of treatment
Traditional Medicine Modern Medicine
Disease can be due to supernatural Disease can be due to

causes arising from the displeasure of physiological imbalance,
God, evil spirit, effect of witchcraft or pathological agents and toxic
the intrusion of an object into the body substances
Based on cultural beliefs Concept based on experiments
and results 10
Basic Principles of Ayurveda
Ayurveda is based on the premise that the universe and our body
is made up of five elements: Air, Fire, Water, Earth, Ether
These elements are represented in humans by three "doshas“ or

energies:
'Tridosha' or the Theory of Bio-energies
1. Vata relevant to air and ether elements. This energy is controls

respiration, nerve impulses, circulation, and elimination.
2. Pitta relevant to fire and water elements. This dosha controls

metabolism.
3. Kapha relevant to water and earth elements. Kapha controls

growth and protection. 11
• When any of the doshas accumulate in the body beyond the
desirable limit, the body loses its balance.
• Every individual has a distinct balance and able to restore this.
• Ayurveda suggests specific lifestyle and nutritional guidelines to
help individuals reduce the excess dosha.
A healthy person, as defined in Sushrut Samhita whose:

• doshas are in balance
• appetite is good
• all tissues of the body and all natural urges are functioning
properly.
• mind, body and spirit are cheerful" 12
Diagnosis: Ayurvedic practitioners (Baidyas) approach diagnosis by
using all five senses.
• Observation of breathing and speech.
• The study of the lethal points of special importance through
pulse diagnosis.
Treatments: Usually given in one of two forms:

1. Panchakarma: A 5-different body purification program
2. Use of herbal and other natural products (animals/ animal

products and minerals) as medicine to balance the body
13
THE SIDDHA SYSTEM OF HEALTH AND MEDICINE
• Siddha medicine is the oldest medical system in the
world.
• Siddha is a Tamil word derived from siddhi-one who has
attained perfection in life or heavenly bliss.
• Practitioners of this system were called siddhars.
• Wise men who meditated, wrote poems and had
healing powers, siddhars were originally devotees of
Lord Shiva.
• In ancient India, 18 important siddhars developed the
system, which is why it's called Siddha medicine.
• It evolved and flourished in south India (Dravidian
culture).
• The Siddha system is based on the principle that the
macrocosm (the universe) and the microcosm (man)
are similar.
• Man is made up of five fundamental elements: earth
(solid matter), water (liquid matter), fire (energy), air
(gaseous matter) and ethereal space between the
other four elements.
• Reflecting this theory of cosmic oneness, the five
senses of man are said to correspond with the five
elements.
• Ether (akasam) is responsible for hearing; fire
(theyu) for sight; air (vayu) for smell; water (appu)
for taste; and earth (prithvi) for the sense of touch.
• Herbal, animal or inorganic chemical compounds,
such as sulfur and mercury, used as therapies for
treating diseases (balancing the elements).
COMMON MEDICINAL PLANTS/ ANIMALS/ MINERALS USED IN
AYURVEDA AND SIDDHA
Acacia catechu Glycyrrhiza glabra Emblica officinalis
Curcuma longa Hibiscus rosa - sinensis Terminalia chebula
Cymbopogon citratus Brassica nigra Terminalia bellerica
Datura stramonium Santalum album Aegle marmelos
Papaver somniferum Saraca indica Adhatoda vasica
Ficus bengalensis Azadirachta Indica Acorus calamus
Oryza sativa Moringa oleifera Terminalia arjuna
Aconitum ferox Commiphora mukul Honey
Aloe barbadensis Commiphora myrrha Kaolin
Piper longum Zingiber officinale Gold
Ferula foetida Atropa belladona Silver
Plantago ovata Triticum aestivum Musk
Foeniculum vulgare Withania somnifera Ghee
• Unani medicine originated in Greece or unan.
• Greek philosopher-physician Hippocrates (460-377BC)
• After Hippocrates, a number of Greek scholars enriched the system
and it imbibed the best, from contemporary systems.
17
Principle of Unani Medicine
• The fundamental principle of the Unani system recognizes that disease is
a natural process and symptoms of a disease are body's reaction to
disease.
• The Unani medicine is based on Humoral theory (relating to bodily
fluids), which pre-supposes the presence of four humors:

1. Dum (blood)
2. Balgham (phlegm)
3. Safra (yellow bile)
4. Sauda ( black bile).
• The body has the power of self preservation to maintain a correct
balance of these humors, which is called as Quwwat-e-Mudabbira
• Unani medicines help the body to regain this balance. 18

Diagnosis
The Unani Physician-called Hakim diagnoses a disease by examining:
• Nabz (pulse) and rhythmic expansion of arteries by fingers
• Urine and stool
Treatment
Four types of treatment lines are available:
(1) ILAJ BIL TADBEER (REGIMENTAL THERAPY),
(2) ILAJ BIL GHIZA (DIETO-THERAPY),
(3) ILAJ BIL DAWA (PHARMACOTHERAPY)
(4) JARAHAT (SURGERY) 19
Different Unani medicines includes:
1. Madar 7. Sana
2. Fufal 8. Tagar
3. Gilo 9. Zeera
4. Kabab chini 10. Lodh
5. Karanj 11. Quest
6. Kulthi
Single drugs or their combinations in raw form are preferred over

compound formulations 20
• Homeopathy derived from the Greek words hómoios= "like-"
+ páthos "suffering").
• It is a system of alternative medicine originated in 1796 by Dr.

Samuel Hahnemann, based on the doctrine of similia
similibus ("like cures like").
• According this, a substance that causes the symptoms of a disease in

healthy people will cure that disease in sick people.
• Homeopathy based on vitalist philosophy.
• Homeopathy maintains the vital force, which has the ability to react
and adapt to internal and external causes, refer as the "law of
susceptibility”. 21
Drugs in small doses induce symptoms
the artificial symptoms stimulates the vital force
stimulated vital force neutralize and expel the original disease.
Homeopathic practitioners rely on two types of reference when

prescribing remedies:
1. Materia medica: collection of "drugs", organized

alphabetically.
2. Repertories: An index of disease symptoms that lists
remedies associated with specific symptoms.
22
Plant, animal, mineral and some synthetic substances used by
homoeopaths
Plants Arnica, Belldona, Marigold, Colchicom, Hemlock,
Lycopodium, Opium, Aconite, Thuja, Nux-vomica, Ergot
and Hyoscyamous etc.
Animals Honey-bees and Cantharides etc.
Minerals Arsenic oxide, Barium carbonate, Calcium phosphate,
Mercuric chloride, Antimony tartarate, Sulphur, Copper,
Aluminium, Phosphorous and Platinum etc.
Homeopaths also use:

• remedies prepared from biological matters of healthy and
diseased bodies of human or animals
23
• A solution that is more dilute is described as
having a higher potency.
• “Dynamisation" or "potentiation“: Drug is

diluted with alcohol or distilled water and
then vigorously shaken by 10 hard strikes
against an elastic body, a process homeopaths
also call as "succussion“.
24
• Traditional Chinese Medicine (TMC) is a broad range of medicine
practices.
• These are based on the tradition of more than 2,000 years.
• These includes:
▪ Chinese herbal medicine
▪ Acupuncture
▪ Massage (Tuina)
▪ Exercise (Qigong)
▪ Dietary therapy
▪ Tai chi
25
The ancient beliefs, on which TCM is based include the followings:
• The human body is a miniature version of the larger universe.
• Five elements—fire, earth, wood, metal, and water—symbolically
represent the stages of human life, explain the functioning of
the body during health and illness.
• Balance between two opposing yet complementary forces, called
yin (cold/water) and yang (heat/fire) responsible for good
health.
• Qi, a vital energy that flows through the body. Free flow of this
energy is necessary for good health. 26

Diagnosis aims to trace an underlying disharmony:
• by measuring the pulse, inspecting the tongue, skin, and eyes
• looking at the eating and sleeping habits of the person as well as
many other things.
Treatment includes the use of herbs and herbal formulations:

• TCM identifies and recommends drugs believed to treat imbalance
of yin and yang.
• Over 12,800 types of Chinese medicinal agents are used:
❖ Over 11,000 medicinal plants
❖ Over 1500 medicinal animals
❖ Over 80 medicinal minerals.
The doctrines of Chinese medicine are mentioned in books such as

1. The Hung-Di Nei-Jing (Yellow Emperor's Inner Canon)
2. The Treatise on Cold Damage. 27
• The ancient civilizations developed their systems of medicine
(Ayurveda, Unani and Siddha) independent of each other but
all of them were predominantly plant based and comprise over
8000 medicinal and aromatic plants species.
• Plants are the great source of natural chemicals.
• Traditional practitioners use only 7,000–7,500 plants for curing
different diseases.
• The proportion of use of plants in the different Indian systems
of medicine is Ayurveda 2000, Siddha 1300, Unani 1000,
Homeopathy 800, Tibetan 500, Modern 200, and folk 4500.
• In India, around 25,000 effective plant-based formulations are
used in traditional and folk medicine.
• The medicinal herbs assume a tremendous importance at a
time when the whole world is showing a resurgence of interest
in the healing properties of plants.
• The renaissance of medicinal and aromatic plants for health
care is gaining immense popularity in the recent years, not only
in the developing countries but also in developed industrialized
countries.
• These herbs not only provide raw materials for the
manufacture of allopathic drugs but have also served
the hillman for decades and suit his local medicinal
system.
• From among the 2.5 Lakh plant species in the world,
two thousand species are used for medicinal purpose.
• It has been reported that there has been an alarming
increase in number of diseases and disorders caused
by synthetic drugs prompting a switch over to
traditional herbal medicine.
• The rising demand for complementary and alternative
medicine over the past decade is a true gross-root
phenomenon.
• They are also cost effective and gaining wide spread
acceptance for their effectiveness
• A large number of manufacturing units, some
with multi crore investment and some
multinationals, have also entered in this area of
drugs and pharmaceuticals.
• A majority of raw materials, used by these
industries, are plant based and the bulk of these
are made available from forest or other natural/
wild sources through various trade channels but
these, generally lack in uniform quality. This
creates a frightening to the desired therapeutic
efficacy of drugs.
• Pharmacognosy helps here through cultivation
and standardization of herbs.
ROLE OF PHARMACOGNOSY IN FINDING EFFICASY OF NEW PHYTOCOSTITUENTS
• Recently, an Israeli company, Galilee Herbal Remedies, has discovered a
cure for pain caused by migraine headaches using a herb Panacetum
parthenium (commercially known as fever few), that has been known
for its medicinal properties for over 200 years. It not only relieves the
migraine headache but also helps alleviate pain due to arthritis and pre-
menstrual tension.
• The fight against HIV and AIDS may have recruits from India in the form
of an array of plants which have been used in Ayurvedic, Unani and
other traditional systems of medicine.
• Researchers at New Delhi's National Institute of Immunology (NII) have
begun investigating the anti-HIV properties of seven plants, namely,
Tulsi (Ocimumsanctum), Brahmi(Centella sp.), Ashwagandha (Withania
somniferum), Punarnaya (Boerhavia difusa), Satavari(Asparagus
racemosus), Gilogiloya (Tinospora cordifolia), and Nirbrahmi (Bacopa
monniera) for their anti-stress properties,and their ability to stimulate
the immune system.
• NII researchers are also excited by the results of experiments with
Neem (Azadirachta indica). Extract from the Neem leaf inhibited HIV in
vitro. Elucidation of Neem's anti-HIV properties was a spin-off from
researches at NII about the plant's use in birth control
ROLE OF PHARMACOGNOSY IN FINDING EFFICASY OF NEW PHYTOCOSTITUENTS
• Reports from the southwest Cameroon in Africa reveal a

rainforest vine that has shown are markable activity
against HIV-virus at least in-vitro.
• The National Cancer Institute (NCI) of USA is doing
intensive experimentation with the michellamine B, the
active bio-compound of woody vine, a species previously
unknown to the western science.
• The NCI program is the largest in the world, based on
collection of plants in 25 developing countries, to screen
natural products for anti-cancer and anti-HIV activity.
• Since 1987, they have tested some 7000 species and
stored hundreds of thousands of natural extracts.
• Since the Rio Earth Summit, many third world countries
are moving to control access to genetic resources
particularly in view of their immense therapeutic
potential.
Prof. I. P. Padhy
Tissue culture
Concept
from a single cell.
1896
differentiate
Totipotency
History
1920
root of tomato



plant cells
Stages

1

Functions
Water
Growth regulators

Organic compounds


Minerals
Energy source
and carbon
Vitamins
Growth regulators
abscisic acid etc.
Gelling agent Agar

Stock Solution

______________________________________________________________________________________________________________________________________________________________


Aseptic Plant



Types of Culture
1. Callus Culture
auxin and cytokinin

(Callus tissue)




medium.
Curve
Total cell count
Packed cell volume

3. Root Cultures
from explants of

5. Embryo Culture
embryos.

technique,





Protoplast Fusion

forced to fuse.

callus.
Plant Regeneration
Organogenesis
Applications

precursors.

culture.

aseptic plants
….Applications
6 Plant breeding

are called clones.



phyto-chemicals.
……Applicat
ions
steps. Example:



of Ginkyo biloba
Phytoconstituents
includes:
2. Rosmarinic acid
3. Opium alkaloids
4. Ginsenosides
5. Ajmacillin
condition
secondary
Cell suspension
producing plants
yield
Product
recovery
Purification
Specific product
Phytoconstituents
hairy root culture
metabolites in
shoot culture

phytoconstituents

pepsin).

Fat, Bees Wax.
Dr. I. P. Padhy
animals.
cellulose etc.).
E. Gums: Guar gum
Seaweed Gum: Agar.


sodium tartrate.
red precipitate.

precipitate.
TEST FOR GUMS

TEST FOR MUCILAGES

red.


Description:
on cooling.
constituents
Chemical Tests:
prolonged heating.
Uses:


Macroscopy:
12 x 2 mm.
rough.
deep yellow ppt.
warmed
to moderate yellow)
Uses:
cream preparations.
Leguminosae.
Morocco and Africa.
making incision.
Macroscopy:
Odour: Odourless
tears.
diameter
minute fissures
95 % alcohol.
Uses:

emulsifying agent.

HONEY


Zealand.
water.
honey comb.
fermentation.
Morphology

sometime.

fragrance.
Uses
mucous membrane.
24
derivatives.”
O
H2O O
+
26
27
• Greasy
benzene.
28
oil”.
29
rancified oils).
1 gm of the oils.
– Viscosity
30
produced
Castor oil

very viscous liquid
• Taste: Acrid
Castor seeds
32
applications.
33
acid is present.

Uses:
34
Preparation:
bags.
or chlorine.
35
Description:
Uses:
36
Lanolin/ WOOL FAT
lanae; Laniol.
Preparation:
water.
acid.
with reagents.
Characteristics
• Yellowish white
Uses
CHAULMOOGRA OIL

Flacourtiaceae.
Indo-China.
Characteristics:
Uses
therapeutic use.
GELATIN

Characteristics
bacteria.
strength).
histidine (0.8%).
Chemical Tests:
give this test.
Uses
texturizer in food.
CASEIN
Biological Source:
calves.
Characteristics:
Uses:
Enzymes
life processes.
heat or cold
side of the cell.
phosphorylases.
enzymes [amylases].
vertebrates.
PAPAIN

PREPARATION:
artificial heat.
alcohol.
PROPERTIES:
powder.
Other Characters:
CHEMICAL TEST
USES:
contact lenses.
PEPSIN
PREPARATION:
• Filter it
half saturated.
in the solution.
DESCRIPTION:
Other characters:
USES:
SERRATIOPEPTIDASE
Biological Source:
Characteristics:
in stomach.
Uses
India.
antibodies.
UROKINASE
Preparation:
drying.
Characteristics:
as SCUPA.
Uses:
catheters.
STREPTOKINASE

S. griseus.
Preparation:
Characteristics:
Uses:
conformation.
clots.
artery thrombosis.
BROMELAIN
Taiwan.
Characteristics:
dormancy.
develop.
compounds.
calcium.
Uses
the infection.
thrombophlebitis.
B. Pharm. APPLICATION OF COMPUTERS IN PHARMACY 2 nd Sem.
APPLICATION OF COMPUTERS IN PHARMACY

Drug Information storage and Retrieval
The Storage, retrieval and dissemination of information constitute a major function of any drug
information service center. Computers play a main role and support for pharmaceutical information
services. Various computer programs have been developed and available to aid the drug information
specialist in searching and retrieval of drug information.
Complete search of the drug information is necessary for the doctor’s, pharmacist’s, nurse’s and
even general public to satisfy the queries about pharmacological actions, drug interactions, adverse drug
reactions, toxicology, etc.
Computerized retrieval of information from large medical databases, is an area of Medical
Informatics which has been rapidly expanding. For retrieval of medical information some international
data banks are available such as Excerpta Medica, MEDLARS and DIALOG.
Excerpta Medica is an information system that provides abstracts of more than 25,00,000 items in
a database that covers more than 5000 biomedical journals. It provides descriptive indexes of biomedical
and clinical literature. MEDLARS ( Medical Literature Analysis and Retrieval System ) is one of the best
and most widely used computer based retrieval service by National Library of Medicine. MEDLARS
systems locates medical and scientific articles not only by keywords and content but also by author. The
GTE Telnet Medical Information Network and the American Medical Association (AMA) have collaborated
to produce several databases as AMA/NET Drug Information Base which includes standard dosages,
action and interactions and information of special patient cases for many drugs. AMA/NET Disease
Information base contains descriptions and diagnostic symptoms for many diseases.
Pharmacokinetics
Pharmacokinetics is a science which deals with the rate of absorption, metabolism, distribution
and elimination of drug and its metabolites in the body. It analysis is basically carried out to get
information on renal clearance, volume of distribution, metabolic disposition, absorption and multiple
dosing of drugs etc. This analysis involves non-linear least square regression analysis. This involves
fitting the experimental data to a set of differential equation and selects the parameters with acceptable
goodness of fit.
Computers simplify tedious calculations and allow more time for the development of new
approaches to data analysis and pharmacokinetics modelling. Computer software’s are used for
development of experimental study designs, statistical data treatment, and data manipulation, graphical
representation of data, pharmacokinetic model simulation, and projection or prediction of drug action.
Mathematical Model for Drug Design
Drug design involves in development of drug molecules on rational basis. This reduces the
expenditure of research in development of drug moiety. Quantitative Structure Activity Relationship
(QSAR) is one of the methods in drug design. QSAR transforms the chemical structure of a compound
into a set of numerical descriptors of the properties relevant to the biological activity and establish the
quantitative relationship between these descriptors and biological activity. The computerised QSAR
approach will enable one to predict the activities of the analogs of a moiety prior to their synthesis. But
these regression methods only permit optimization of a known family of compounds. These do not allow
one to go beyond the common chemical frame of the particular family analysed.
So to overcome this limitation, the molecular modelling technique was developed. The
computerised molecular modelling systems widen the molecular models possible. Molecular modelling
can be carried out by (i) Theoretical Calculation and (ii) Computer Graphics. The molecular modelling
systems based on molecular graphics has facilitated manipulation and visualisation of isolated or
interacting three dimensional ( 3-D) dynamic structures. This molecular modelling technique allows study
of as yet unknown molecules. This 3-D method not only allows manipulation and visualisation but also the
calculation of additional structural properties associated with the molecular structure such as volume,
energies, surface charge densities etc. The various software for this technique are MMMS, SYBYL,
MMSX, AMBER, SCRIPT, etc.
RCPHS 1 By JKC
Hospital Pharmacy
Hospital pharmacy refers to a place within the hospital where all the drugs and medications are
stored in order to provide in-house treatment to patients those have been admitted in that hospital.
Some of the Functions of Hospital Pharmacy

∑ Ensure purchase and proper storage of drugs.
∑ Label all drugs containers properly.
∑ Ensure proper storage conditions as cold storage and refrigerator.
∑ Dispense drugs as per prescription.
∑ Discard expire medicines and order for fresh stock.
∑ Ensure timely stock audit and its provision.
∑ Maintain proper records related to drugs, their manufacturing and expiry, batch, etc.
∑ Provide support in research and training programs organized within the hospital.
∑ Ensure availability of right medication at the right time and at minimal cost to the patients.
∑ Act as counsellors between doctors, nurses and attendants of patients.
Clinical Pharmacy
Clinical pharmacy refers to study of science dealing with best utilization of pharmacist’s
experience and knowledge to provide safest medication during the course of effective patient care.
Some of the Functions of Clinical Pharmacy

∑ Ensure maintenance of complete patient record like drug reactions, allergies, hypersensitivity, etc.
∑ Save physician’s time and effort to decide best treatment for any patient.
∑ Train and assist medical supervisor or physician about various drug complications, interactions,
dosages, efficacies etc.
∑ Assist physician in selecting best drug therapy for a particular patient.
∑ Keep a close track of any drug reactions or contra-indication among the patients.
∑ Handle medical emergency in case of overdoes or poisoning by providing best available antidote.
∑ Assist in discharge counselling related to OTC ( over the counter ) drug medications as per
patient’s response to undergoing drug therapy.
Application of Computer in Hospital & Clinical Pharmacy :

Some of the important points are :
∑ Maintain and assess of patient records.
∑ Maintain and assess drug records.
∑ Evaluate stock and its timely updation.
∑ Discard expiry and timely expiry medications.
∑ Ensure proper and timely supply of medications.
∑ Review contra medications, overdose or any other adverse reactions among the patients.
∑ Details study on various medications available in pharmacy.
∑ Attend online research and training programs.
Electronic Prescribing and Discharging the Medicines

Electronic prescription refers to computer based medical prescription which is generated online. It
has replaced use of pen and paper. It allows nurses, pharmacists and patient’s attendants to clearly look
into the prescription.
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Benefits of Electronic Prescription
1) Error free dispensing.
2) Automated and faster refill on ongoing treatment.
3) Track any overdoes, drug interactions or allergies’.
4) Track whether a prescription has been refilled or not.
5) Provide better record maintenance services as detailed information about the patient is
available right from the start of the treatment.
6) Reduce chances of self medication and overdoses.
7) Keep a track of prescription related to controlled substances or narcotic drugs.
Barcode Medicine Identification :
The Barcode Medicine Administration is an inventory control system that uses barcodes to
prevent human errors in the distribution of prescription medications at hospitals. The goal of Bar coded
Medication Administration is to make sure that patients are receiving the correct medications at the
correct time by electronically validating and documenting medications. The information encoded in
barcodes allows for the comparison of the medication being administered with what was ordered for the
patient. The barcode medicine identification dispensing has been formulated to ensure 5 fundamental
rights of the patient. They are
∑ Right Medicine
∑ Right Patient
∑ Right dose
∑ Right time
∑ Right mode of administration
This system of dispensing has also reduced time gap between actual prescription and dispensing.
Further, it is useful for managing inventory as well as billing. Barcode dispensing is faster, easier, more
manageable and error free mode of dispensing medications.
Automated Dispensing
Technological advancement are a constant in today’s health care marketplace where payers and
patients demand high quality, efficient and cost-effective service. Improving patient safety is always a key
focus in the hospital setting and pharmacists have been exploring a variety of strategies and technologies
to achieve this goal. Automated dispensing machines-decentralized medication distribution systems that
provide computer controlled storage, dispensing and tracking of medications have been recommended as
one potential mechanism to improve efficiency and patient safety and they are now widely used in many
hospitals.
Automated Dispensing Systems (ADS) are variously described as automated dispensing cabinets,
automated dispensing devices, automated distribution cabinets, and automated dispensing machines and
dispensing robots. These computer controlled devices are designed to securely store, dispense and track
medications and as a result reduce the occurrence of medication errors. Improvements in workflow and
cost efficiencies are also expected as a result of reduced staff time requirements, improved storage
capacity and stock control and so on. Robot use in community pharmacy is still relatively limited.
However, robots have the potential to handle high volumes of dispensing in community pharmacies, or
dispensing “hubs”, and to release pharmacists to develop and deliver patient-centred services.
Mobile Technology and Medication Adherence
Despite the existence of many effective medical treatments, there is a great difference between
the outcome projected results and actual outcome results of medication. This gap has been attributed
partly to lack of patient adherence to recommended treatment. The low medication adherence rates to :
∑ Poor communication between the drug provider and patient
∑ Socioeconomic barriers for the patient
∑ Complexity of medication regimen prescribed
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If left unchecked, the high prevalence of medication non-adherence may have a substantial and
detrimental impact on pollution health and economy. There are a number of methods used to measure
medication adherence and these methods are categorized as either direct or indirect.
Direct Methods : Taking medication in the presence of a health care provider, measuring the level of
medicine or metabolite in blood or measuring biological markers in blood, once a prescription is taken.
In-Direct Methods : Indirect methods include : pill counts, self-report, patient diaries, pharmacy refill
data, electronic medication monitors, and other telemedicine devices.
Direct measures are considered by some to be more reliable and accurate than indirect
measures, most agree that direct methods are too burdensome and not practical for routine clinical use.
Human interventions that once required direct patient contact can now be performed electronically
using Mobile technology ( Health IT ). For instance, with the advent of video conferencing and
Smartphone applications, patients and health care providers are now able to monitor medication
adherence through video-logged confirmation of dosage. Using mobile technology increases the
feasibility of monitoring medication adherence and promotes technological innovations that are
consumer-facing to improve self-management.
Electronic Drug Monitoring
Self reporting is the most common method for calculating medication adherence. Innovations in
health IT have accelerated the use of electronic drug monitoring systems (EDMs) to measure adherence.
EDMs use monitoring devices, such as medication event monitoring systems (MEMS), a pill bottle cap
embedded with a microprocessor that records the date and time of opening of each bottle as a
presumptive dosage. Other advances include electronic pill boxes, or tracing pharmacy refill data and
sending automated alerts to the prescriber, when the medication is not filled.
Substitutable Medical Apps, Reusable Technologies ( SMART ) Platform
The SMART platform’s goals are to develop application programming interfaces, that enable
compliant Electronic Health Record (EHR) technologies to interface with multiple third party-developed
medical applications. This can be compared to cell phone carriers creating a set of APIs that enable a
smart-phone to run on multiple third-party apps.
Diagnostic System
Medical diagnosis ( abbreviated Dx or Ds) is the process of determining which disease or
condition explains a person’s symptoms and signs. The information required for diagnosis is typically
collected from a history and physical examination of the person seeking medical care. Often, one or more
diagnostic procedures, such as diagnostic tests, are also done during the process. Now a day’s
computer-based system designed to support clinical decision making. These systems are generally
designed to provide efficient access to medical information, they also include mechanisms for the
assessment of clinical and laboratory data and the provision of diagnostic advice.
Lab Diagnostic System
A medical laboratory or clinical laboratory is a laboratory where tests are usually done on clinical
specimens in order to obtain information about the health of a patient as pertaining to the diagnosis,
treatment and prevention of disease. A diagnosis based significantly on laboratory reports or test results,
rather than the physical examination of the patient. A proper diagnosis of infectious diseases usually
requires both an examination of signs and symptoms, as well as laboratory characteristics of the
pathogen involved.
The laboratories provide testing for diagnosis, surveillance and treatment monitoring at every
level of healthcare system of patients. A Laboratory Information System (LIS) or Laboratory Management
System (LMS), is a software-based laboratory and information management system with features that
support a modern laboratory’s operations.
Patient Monitoring System ( PMS )
Patient monitoring is a process of collecting medical parameters of a patient. It is a very critical
monitoring systems, used for monitoring physiological signals including ECG, Respiration, Invasive and
Non-Invasive Blood Pressure, Oxygen Saturation in Human Blood, Body Temperature and other Gases
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etc. It is not possible for any doctors to attend a particular patient throughout the days. By implementing
PMS, it recorded all the vital parameters of the patients by putting sensors at the different parts of the
body. It recorded all the vital parameters as well as display on the screen. When the system found any
abnormal reading than immediately gives signal to the doctors as well as sisters room as an indication to
attend the patient immediately so that the valuable life of the patient can be saves.
Pharma Information System
Pharma Information System refers to use of information technology in the field of pharmaceutical
industry. The science of technology that deals with storage, retrieval and use of information related to
medical industry and pharmaceutical drugs is known as Pharma Information System.
Benefits of Pharma Information System
1) Faster Access
2) Easier to use
3) Error free
4) High reach to the people
5) Expert advice
6) Safer practice
7) Increased efficiency
8) Reduced cost
9) Increased knowledge
10) Qualitative assessment
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B. Pharm. BIOINFORMATICS 2 nd Sem.
Definition of Bioinformatics :
Bioinformatics is a branch of science that deals with the study of biological information by using
computer technology. Computers are used to gather, store, analyze and integrate the biological information
which can then be applied to the gene based drugs discoveries and development.
Scope & Objectives of Bioinformatics :
Bioinformatics is defined broadly as the study of the inherent structure of biological information. It is
the combination of biology and the information sciences. Examples of current bioinformatics research
include the analysis of gene and protein sequences to reveal protein evolution and alternative splicing, the
development of computational approaches to study and predict protein structure to further understanding of
function, the analysis of mass spectrometry data to understand the connection between phosphorylation
and cancer, the development of computational methods to utilize expression data to reverse engineer gene
networks in order to more completely model cellular biology, and the study of population genetics and its
connection to human disease.
Graduates in bioinformatics can expect to engage in any combination of research, teaching, clinical
service, and consultation. Within universities and research centres there is a growing need for
bioinformatics researchers who can analyze new sources of high-throughput experimental data in biology,
medicine, and bioengineering. Biotechnology and pharmaceutical companies also seek bioinformatics
graduates for applied research on disease — and drug discovery. Medical centres are also increasingly
hiring bioinformatics graduates as genomics data become important in medical research and clinical
applications.
Role of Bioinformatics in Pharmaceutical Research
In pharmaceutical research, bioinformatics typically equates to the discovery of novel drug targets
from genomic and proteomic information. Bioinformatics can be subdivided into several complementary
areas like gene informatics, protein informatics, system informatics, etc.
Gene informatics, with links to genomics and microarray analysis, is concerned, inter alia, with
managing information on genes and genomes and the in silico prediction of gene structure. A key
component of gene informatics is gene finding: the relatively straight forward searching, at least
conceptually if not always practically, of sequence databases for homologous sequences with hopefully
similar functions and analogous roles in disease states.
Protein informatics concerns itself with managing information on protein sequences and has
obvious links with proteomics and structure-function relationships. Part of its remit includes the modelling
of three-dimensional structure and the construction of multiple alignments.
System informatics component concerns itself with the higher-order interactions rather than
simple sequences and includes the elaboration of functional protein-protein interactions, metabolic
pathways and control theory.
Bioinformatics is managing the information generated by microarray experiments and proteomics
and drawing from it data on the gene products implicated in disease states. Bioinformatics is still largely
concerned with data handing and analysis, be that through the annotation of macromolecular sequence and
structure databases or through the classification of sequences or structures into coherent groups.
Goals of Bioinformatics
1) Bioinformatics organizes data in a way that allows researchers to access existing information and to
submit new entries as they are produced. The purpose of bioinformatics extends for beyond mere
volume control of data. For example : GenBank for nucleotide and protein sequence information,
Protein Data Bank for 3D macromolecular structures, etc.
2) To develop tools and resources that air in the analysis of data. For example : BLAST to find out
similar nucleotide / amino-acid sequences, ClustalW to align two or more nucleotide / amino-acid
sequences, Premer3 to design primers probes for PCR techniques, etc.
3) Bioinformatics is to exploit these computational tools to analyze the biological data interpret the
results in a biologically meaningful manner.
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Bioinformatics Databases
A database is a computerized archive used to store and organize data in such a way that
information can be retrieved easily. Biological databases comprise not only data, but also sophisticated
query ability and bioinformatics data analysis tools hence, the termed as ‘Bioinformatics Databases”.
Biological databases can be broadly classified into sequence, structure and functional databases. Nucleic
acid and protein sequences are stored in sequence databases and structure databases store solved
structures of RNA and proteins. Functional databases provide information on the physiological role of gene
products, for example enzyme activities, mutant phenotypes, or biological pathways.
Biological Databases
Sequence
Structure Other
Protein Nucleic Acid RELIBASE

PCB
SCOP REBASE
CATH GPCRDB
SwissProt Gen Bank
PIR DDBJ
TrEMBL EMBL
PROSITE
PFAM
List of some frequently used databases :
Database URL Feature
GenBank http://www.ncbi.nlm.nih.gov/ NIH’s archival genetic sequence database
EMBL http://www.ebi.ac.uk/embl/ EBI’s archival genetic sequence database
DDBJ http://www.ddbj.nig.ac.jp/ NIG’s archival genetic sequence database
SGD http://www.yeastgenome.org/ A repository for baker’s yeast genome and biological data
It provides access and statistics for the completed

EBI genomes http://www.ebi.ac.uk/genomes/
genomes
Database that maintains automatic annotation on selected
Ensembl http://www.ensembl.org/
eukaryotic genomes
Each UniGene cluster contains sequences that represent a
UniGene http://www.ncbi.nlm.nih.gov/sites/entrez?db=unigene
unique gene, as well as related information.
Division of GenBank that contains expression tag
dbEST http://www.ncbi.nlm.nih.gov/dbEST/
sequence data
Table 1 : Summary of Nucleotide sequence databases

Description of the function of a protein, its domains structure, post-
Swiss-Prot/TrEMBL http://www.expasy.org/sprot/
translational modifications etc,
UniProt http://www.pir.uniprot.org/ Central repository for PIR, Swiss-Prot, and TrEMBL
It strives to be comprehensive, well-organized, accurate, and
PIR http://pir.georgetown.edu/
consistently annotated.
Pfam pfam.sanger.ac.uk/ Database of protein families defined as domains [49]
PROSITE www.expasy.ch/prosite/ Database of protein families and domains
Table 2 : Summary of Protein sequence databases
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PDB www.rcsb.org/pdb/ Protein structure repository that provides tools for analyzing these structures
Classification of protein 3D structures in a hierarchical scheme of structural
SCOP scop.mrc-lmb.cam.ac.uk/scop/
classes
CATH www.cathdb.info Hierarchical classification of protein domain structure
NDB http://ndbserver.rutgers.edu/ Database housing nucleic acid structural information
Table 3: Summary of Structure databases.
KEGG http://www.genome.jp/kegg/ Protein structure repository that provides tools for analyzing these structures
Classification of protein 3D structures in a hierarchical scheme of structural
BioCyc http://www.biocyc.org/
classes
BRENDA http://www.brenda-enzymes.org/ Hierarchical classification of protein domain structure
EMP http://emp.mcs.anl.gov/ Database of Enzymes and Metabolic pathways public server
BRITE http://www.genome.jp/kegg/brite.html Biomolecular Relations in Information, Transmission and Expression
Table 4: Summary of Pathway databases.
Concept of Bioinformatics
Bioinformatics has emerged as a new branch of biotechnology, offering a fundamental tool to the
biologist to accelerate commercialization of biotechnology. Bioinformatics has been the most powerful tools
for data mining in life science, analysis, data searching, integration and simulation of molecular biological
data. Bioinformatics can be defined as, “the use of information technology to acquire, store, manage,
share, analyse, represent and transmit genetic data.” The ultimate goal of the field of bioinformatics is to
create a global perspective from which underlying principles in biology can be discerned. It thus enables
the discovery of scientific principles upon which biological systems are built, and promotes innovative
discoveries using logical conclusions arrived at from true and reliable data.
Advantages of Bioinformatics
1) Bioinformatics saves a lot of time as computers respond rapidly, give quick results and draw
conclusions faster when compared to results obtained through laborious and time-consuming
laboratory procedure.
2) It saves a lot of money, manpower and financial resources because the computers that it relies on for
performing the work are more efficient and accurate than the individuals and are comparatively
inexpensive to maintain.
3) Complex sequences of nucleotides and amino acids, which cannot be comprehended by the human
brain can be compared and analysed by bioinformatics tools that align segments of the corresponding
sequences to identify matches, mismatches, similarities, gaps, substitutions, etc.
4) Anyone who knows the subject can do bioinformatics work in the comfort of his / her home, work or
anywhere else where availability of Internet.
5) Bioinformatics saves a lot of space or infrastructural facilities because the computers, servers and
machinery required are small with “small foot prints” and hence require little space.
6) It is a predictive as well as a retrospective tool. Advancements in bioinformatics tools based on
Artificial Neural Networks ( ANN), learning software and computational biology enable logical and
accurate predictions.
7) It is highly versatile as the virtual experiments can be repeated at different experimental conditions
without the need for sophisticated analytical machinery or messy and wet laboratory procedures.
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Limitations of Bioinformatics
1) It depends on experimental science to produce raw data for analysis.
2) In bioinformatics many accurate but exhaustive algorithms cannot be used because of the slow rate
of computation.
3) Its predictions are not formal proofs of any concepts. They do not replace the traditional experimental
research methods of actually testing hypotheses.
4) It is by no means a mature field. Most algorithms lack the capability and sophistication to truly reflect
reality. They often make incorrect predictions that make no sense when placed in a biological context.
5) In bioinformatics sequence data from high throughput analysis in often contain errors. If the
sequences are wrong or annotations incorrect, the results from the downstream analysis are
misleading as well. That is why it is so important to maintain a realistic perspective of the role of
bioinformatics.
6) The quality of bioinformatics predictions depends on the quality of data and the sophistication of the
algorithms being used.
Impact of Bioinformatics in Vaccine Discovery
The vaccines development cycle is initiated by activities that deal with the discovery of new
medicinal lead structures. Vaccines are the most effective method to prevent and control the spread of
infectious disease. Bioinformatics, as a newly developed and still emerging field, incorporates a variety of
scientific endeavours, including, but not limited to, the computational analysis of DNA sequence data,
laboratory methods and determine the expression and cellular location of proteins, analysis of transcripts
expressed at different times and under different conditions, computer analysis of genetic and amino acid
sequences for prediction of protein function.
The success of vaccination is reflected in its worldwide impact by improving human and veterinary
health and life expectancy. The invaluable role of traditional vaccines to prevent diseases, the society has
observed remarkable scientific and technological progress, in the improvement of these vaccines and the
generation of new ones. This has be possible by the fusion of computational technologies with the
application of recombinant DNA technology, the fast growth of biological and genomic information in
database banks. This has aided in expanding the concept and application of vaccines beyond their
traditional immunoprophylactic function of preventing infectious diseases. This technology also serving as
therapeutic products capable of modifying the evolution of a disease and even cure it.
Vaccines are the pharmaceutical products that offer the best cost-benefit ratio in the prevention or
treatment of diseases. Vaccine development and production are costly and it takes years for this to be
accomplished. Several approaches have been applied to reduce the times and costs of their development,
mainly focusing on the selection of appropriate antigens or antigenic structures, carriers and adjuvant. One
of these approaches is the incorporation of bioinformatics methods and analyses into vaccine development.
At present, there are many alternative strategies to design and develop effective and safe new-generation
vaccines, based on bioinformatics approaches through reverse vaccinology, immune-informatics and
structural vaccinology.
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B.Pharm 2 nd Sem
COMPUTERS AS DATA ANALYSIS IN PRECLINICAL DEVELOPMENT
Data analysis is known as analysis of data. It is a process of inspecting, cleansing, transforming and
modelling data with the goal of discovering useful information, suggesting conclusions and supporting decision
making. Preclinical development is a stage of research that begins before clinical trials can be testing and
drug safety gin and during which important feasibility, iterative data are collected. The main goals of
preclinical studies are to determine the safe dose for first-in-man study and assess a product’s safety profile.
Products may include new medical devices, drugs, gene therapy solutions and diagnostic tools.
Initially, all data analysis was performed by using software programs written by each laboratory. The
variety and quantity of software packages grew as newer data analysis techniques emerged. The wide use of
computers has tremendously increased efficiency and productivity in pharmaceutical drug and dosage form
development. Considering the generality of computer applications in every scientist’s daily activities, special
emphases are put on three widely used computer system. They are :
∑ Chromatographic Data Systems (CDS)
∑ Laboratory Information Management Systems (LIMS)
∑ Text Information Management Systems (TIMS)
These three computer systems handle the majority of the work in data / document management in the
preclinical area, supporting the New Drug Application (NDA) and Marketing Authorization Application
(MAA) filings.
CHROMATOGRAPHIC DATA SYSTEMS (CDS)
Chromatography is an analytical technique used in virtually all sectors of the pharmaceutical, medical
device and biotechnology industries to detect or quantify compounds during the course of product
development and manufacture. Chromatography software, known as CDS, collects and analyzes
chromatographic results delivered by chromatography detectors. Many chromatography software packages are
provided by manufacturers, and many of them only provide a simple interface to acquire data. They also
provide different tools to analyze this data. CDS helped the pharmaceutical industry to increase efficiency and
productivity by automating a large part of pharmaceutical analysis and the main focus of CDS has been on
providing accurate and reliable data.
The importance of CDS is directly related to the roles that chromatography, particularly High-
Performance Liquid Chromatography (HPCL) and Gas Chromatography (GC) play in pharmaceutical
analysis. CDS are also used for several other instrumental analysis technologies such as Ion
Chromatography (IC), Capillary Electrophoresis (CE) and Supercritical Fluid Chromatography (SFC).
BASIC CONCEPTS OF CDS
The Days Before Development of CDS

In early 1970s, chromatographs were relatively inefficient. Chromatographers had to use micro-
syringes for sample injection and stopwatches for measurement of retention times. The chromatograms were
collected with a strip chart recorder. Data analysis was also performed manually. Peak areas were obtained by
drawing a “best fit’ triangle manually for each peak and then using the equation
Area = Base X Height.
At that time, the management of chromatographic data was essentially paper based and very
inefficient. However, compared with the traditional analytical methods, the adoption of chromatographic
methods represented a significant improvement in pharmaceutical analysis. It is especially important for
methods intended for early-phase drug development when the chemical and physical properties of the Active
Pharmaceutical Ingredient (API) are not fully understood and the synthetic processes are not fully
developed. Therefore the assurance of safety in clinical trials of an API relies heavily on the ability of
analytical methods to detect and quantitate unknown impurities that may pose safety concerns. This task was
not easily performed or simply could not be carried out by classic wet chemistry methods. Therefore, HPLC
and GC established their places as the mainstream analytical methods in pharmaceutical analysis.
As chromatographic methods become more and more important in the pharmaceutical industry as well
as in other industries, practical needs prompted instrument vendors to come up with more efficient ways for
collecting and processing chromatographic data.
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B.Pharm 2 nd Sem
MODERN CDS
Use of server-based computing is only one of the important features of the modern CDS. Server-based
computing uses a multiuser operating system and a method for distributing the presentation of an application’s
interface to a client device. There are no software components installed on the client PC. The client’s PC
simply acts as the application server’s display. The other two important features of modern CDS are the use of
embedded data structure and direct instrument control. The earlier generations of CDS used a directory file
structure, meaning that the raw data and other files such as the instrument method and data processing
method were stored at separate locations which is problematic and causes data redundancy.
Embedded Data Structures
The embedded relational database has been widely used by LIMS and is a much better file structure.
The embedded data structure can be used to manage not only chromatographic data, but also all aspects of
the CDS, including system security and user privileges. The embedded data structure maintains all information
and changes by date and time stamping them to prevent accidental overwriting of raw data and method files. It
controls versions of all processed result files, acquisition methods, processing methods and reporting methods
to provide full audit trails. All of the metadata (acquisition, process and reporting methods) related to a specific
result are tied together.
Direct Instrument Control
It was an important issue for the earlier version of CDS. The scheme of connecting the detector
channels through analog to digital (A/Ds) to CDS worked well in analytical laboratories across the
pharmaceutical industry. The scheme provided enough flexibility so that the CDS could collect data from a
variety of instruments, including GC, HPLC, IC, SFC and CE. It was equally important that the CDS could be
connected to instruments that were manufactured by different vendors.
Major CDS Vendors and Their Products
Products Vendor URL
Atlas Thermo Electron Co. www.thermolabsystems.com
Cerity Agilent Technologies, Inc. www.agilent.com
Chromeleon Dionex Co. www.dionex.com
Class VP Shimadzu Scientific Inst. www.shimadzu.com
Empower Waters Co. www.waters.com
EZChrom Elite Scientific Software, Inc. www.scisw.com
Galaxie Varian Inc. www.varianinc.com
TotalChrom Perk in-Elmer, Inc. www.perkinelmer.com
LABORATORY INFORMATION MANAGEMENT SYSTEMS (LIMS)

Laboratory Information Management System (LIMS) is a software-based laboratory and information
management system. A comprehensive LIMS specifically designed for a cytogenetic laboratory may include
the ability to collect and store data, analyze and report results, interface with lab instrumentation and outside
clinical facilities, organize workflow, evaluate personnel performance, monitor Quality Assurance (QA) and
Quality Improvement (QI) parameters, ensure accurate billing practice and quality control procedures. LIMS
applications exist on a variety of database platforms such as dBase, Microsoft Access, Oracle and Meditech in
addition to proprietary systems. LIMS emerged to address needs for managing the totality of the laboratory’s
analytical testing data and information. It represent an integral part of the data management systems used in
preclinical development, more specifically in drug substance and drug product stability studies.
Components of LIMS
Components of a LIMS include both computer hardware and software. The system’s physical hardware
components include its computer processor, peripheral devices such as terminals, printers, disc drives etc.
and elements such as cables and switches that connect the various parts. The LIMS application provides
general laboratory functions. The exact functions provided and approach taken can differ significantly from
one LIMS application to another.
A wide variety of other applications software can be on the system. This includes laboratory software
packages for statistics, structural data management, word processing, spreadsheets, textual data
management, image management and document control. An LIMS, consisting of specifically designed
software and properly installed hardware and peripherals, could be customized to facilitate a wide variety of
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B.Pharm 2 nd Sem
laboratory activities. The most advanced LIMS utilize server-based architecture to ensure system security and
control. There are four main types of architectural options when implementing LIMS. They are :
∑ The LAN : In a multiple-site situation and through the standard client /server setup, the application
would be hosted separately on a server at each site connected to PC clients. In this setup, the LIMS are
installed on both the clients and the server. The system administration is required at each facility.
∑ The WAN : In this setup, the LIMS take advantage of telecommunication technology to cover a great
distance. The setup can also be used to connect disparate LAN’s together. In this setup, the LIMS are
installed on both the clients and a central server.
∑ The Centrally Hosted Thin Client : For this setup, system administration is managed at a corporate
centre, where the LIMS are hosted and distributed via a WAN or the Internet with Virtual Private
Networks(VPN).
∑ The ASP Hosted : In this setup, the LIMS are hosted on a centrally managed server form and
maintained by third-party specialists. Users access the LIMS with any Internet-connected PC with a
standard Web browser.
There are large numbers of established vendors that provide commercial LIMS with similar range of
core functionality, but few of them are dedicated to the pharmaceutical industry. Some of them are:
LIMS Vendors SPECIALIZED IN Pharmaceutical Industry
Products Vendor URL
Debra LabLogic Systems Ltd. www.lablogic.com
Q-DIS/QM Waters www.waters.com
QC Client Agilent www.agilent.com
WinLIMS QSI www.lims-software.com
ACD/SLIMS Advanced Chemistry Development www.acdlabs.com
V-LIMS Advanced Technology Corp www.vetstar.com
VET/HEX HEX Laboratory Systems www.hexlab.com
BioLIMS PE Informatics www.pebiosystems.com
LabCat Innovative Programming Assoc www.labcat.com
Advantages of LIMS
1) Efficiency : LIMS streamlines data entry by automating the process. This results in less downtime,
faster access to data, accurate up-to-date data and the ability for the LIMS to grow with the increasing
needs of the lab.
2) Cost Reduction : Total costs of operations such as labour, resources etc. are reduced by using LIMS.
3) Compliance : LIMS can assist in real-time monitoring and quality control. Workflows can be managed,
samples logged, and tests can be checked against protocols and procedures to ensure compliance.
TEXT INFORMATION MANAGEMENT SYSTEMS (TIMS)
TIMS is essential in preclinical development because huge number of text documents and other related
information such as image photographs etc., in the area and requires protection and easy access. It helped
the pharmaceutical industry to improve efficiency in managing business-critical text documents. However it is
still a time-consuming process to write, review, audit, approve and publish text documents for submission.
The objective of an information retrieval system is to organize and store large amounts of text so that
information can be retrieved from the repositories in response to users information requests. A text document
management system is essential in preclinical development because huge numbers of text documents and
other related information are generated in the area. All these documents and data information are considered
intellectual property and require protection and easy access.
Requirement of TIMS
Text retrieval systems deal with various kinds of documents which are textual in nature. The scientists
spend quite a large percentage of their working time on writing compound documents. In today’s environment,
scientist can use a report template to facilitate report writing. Text retrieval systems deal with bibliographic
documents / materials characterised by various keys like author, title, keywords, publication details, abstract,
etc. Such systems enable users to search the bibliographic records through any of the keys like author name,
title, assigned keywords, or by one or more words occurring in the abstract field. In such a situation, the text
retrieval system acts as a reference retrieval system because the search retrieves one or more bibliographic
records, and the users have to consult the hard copies of the document to get the required information.
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B.Pharm 2 nd Sem
Text retrieval systems may also contain full texts of any kind of documents like letters,
correspondences, office memos, legal documents, patient records and case histories, complete texts of
articles and books and so on. Users of such systems expect to retrieve the complete text or parts of it in
response to a query and can go through it to get the desired information. Such systems, because they store
the complete text of documents rather than mere bibliographic information of the same, take much disk space
and the retrieval mechanism needs to be sophisticated to provide fast access to the records.
Documentation Requirements in Preclinical Development
In preclinical development, the Good Manufacturing Practice (GMP) / Good Laboratory Practice (GLP)
regulations are enforced not only for scientific data but also the text documents. The documents managed by
the TIMS are :
∑ The Standard Operating Procedures (SOPs) : The standard operating procedures are controlled in a
way similar to that of specification documents and analytical methods. It must be ensured that the
correct versions of the SOPs are accessed and used by the scientists. After use, the hard copies should
be destroyed and disposed of properly. An added requirement is that the SOPs should be accessible
during working hours without interruption. Hard copies should be available at manageable location so
that the SOPs are available when the electronic system is down.
∑ Research Reports : Research reports such as stability reports, method validation and transfer reports,
and pharmaceutical development reports are key documents used for NDA/MAA filings. These
documents are strictly version controlled.
∑ Laboratory Notebooks : Laboratory notebooks may be debatable to consider laboratory notebooks as
text documents, but they should be mentioned here because of their importance in preclinical
development. Laboratory notebooks are used to record experimental procedures, observations, raw data
and other important information. Currently, most of the major pharmaceutical companies still use paper-
based laboratory notebooks. An Electronic Laboratory Notebook (ELN) is defined by the Collaborative
Electronic Notebook Systems Association (CENSA) as, “a system to create, store, retrieve and share
fully electronic records in ways that meet all legal, regulatory, technical and scientific requirements.”
∑ Product Specification Documents and Analytical Test Methods : Product specification documents
and analytical test methods are important documents and they evolve along with the development
phases. Drug substances and products for clinical trials are tested based on these documents and so
are the stability samples. It is critical to ensure that the analyst will perform the right tests against the
right specifications with the correct version of the test method. Therefore, a mechanism must be in place
to control these documents by using TIMS.
TIMS used in preclinical text document management usually is a simplified version of Enterprise
Content Management (ECM). Various TIMS vendors and their products are given below:
TIMS Vendors and their Products
Products Vendor URL
Document Web Publisher Documentum www.documentum.com
PB WCM FileNet www.filenet.com
TeamSite Interwoven www.interwoven.com
Stellent Content Management Suite Stellant www.stellent.com
V7 Content Management Suite Vegnette www.vegnette.com
Communique Day Software www.day.com
Content Server FatWire www.fatwire.com
Workplace WCM IBM www.ibm.com
Mediasurface Mediesurface www.mediasurface.com
Ingeniux CMS Ingeniux www.ingeniux.com
CommonSpot Paper Thin www.paperthin.com
RedDot CMS RedDotSolutions www.reddot.com
Collage Serena Software www.serena.com
RCPHS 4 By JKC

B.pharm. Class Notes

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B.PHARM.

 Drug metabolism may be defined as the biochemical modification

 It often involves the conversion of lipophilic chemical

Halothane Trifluoro acetic acid

paracetamol N –Hydroxylation derivative

3) It causes conversion of an Inactive drug active metabolite.

 The major site of drug metabolism is the liver (microsomal

 Secondary organs of biotransformation

➢ kidney (proximal tubule)

 Metabolism by organs other than liver (called as extra-

 A few drugs are also metabolised by non-enzymatic means

 The drug metabolising enzymes can be broadly divided

Phase 1 reaction. (Non synthetic phase).

➢ a change in drug molecule. generally results in the

 Addition of oxygen/ negatively charged radical or

removal of hydrogen/ positvely charged radical.

 Reactions are carried out by group of enzyme

monooxygenases in the liver.

 Fianl step: Involves cytochrome P-450 haemoprotein,

NADPH, cytochrome P-450 reductase and O2

 Multiple CYP gene families have been

 Most of the drug metabolizing enzymes are

 CYP3A4 is very common to the metabolism

1A1 Caffeine, Testosterone, R-Warfarin

 Monoamine Oxidase (MAO), Diamine Oxidase (DAO)

➢ MAO (mitochondrial) oxidatively deaminates endogenous

➢ Dopamine, serotonin, norepinephrine, epinephrine

 Alcohol & Aldehyde Dehydrogenase

➢ Non-specific enzymes found in soluble fraction of liver

 Aromatic and side chain hydroxylation.

 Oxidation of benzylic carbon atom

 Oxidation of allylic carbon atom

 Oxidation at the carbon alpha to carbonyl and imino group.

 Oxidatiive N and O-dealkylation

 N- oxidation and sulphur oxidation.

 Hydroxylation means addition of –OH group to the

 The reaction involves the formation of an epoxides as

 oxidative N and O-dealkylation involves hydroxylation of

alpha carbon adjecent to amino group and oxygen to form

carbinolamine or hemiactal as an intermedite than

undergoes clevage to release the alkyl group and the drug

 Nitro,azo and carbonyl group containg drug are easily

 Drugs containing ester or amide functional group

 Drugs containing ester or amide functional group

 Hydrolysis of amide is very slow as compared to

 These are also called conjugation reaction.

 Last step in detoxification reactions and almost always results

 May be preceded by one or more of phase I reaction .

 The products formed after phase-II reactions are generally

 It involve conjugation with various endogeneous substances

 Phase-1 metabolites having free alcoholic or phenolic groups

 This process is done by the help of the enzyme glucuronyl

 Glucuronides formed are normally non toxic,highly polar and

 It mainly occurs in many tissues like kidney,skin,intenstine,lungs

 Also known as mercaptouric acid is a thiol containing

 Electron deficient metabolites like alkyl aryl

 That catalysed by the enzyme glutathione transferase.

 Drug having hydroxy groups,phenols and aromatic amines

 It occurs by the enzyme sulphotransferase and coenzyme

 Mostly the steroidal drugs undergoes by this reaction

 The proces occurs mostly in the liver ,kidney and intenstine.

 Ex-Estrone on metabolism produces estrone sulphate.