Primary metabolites, Fibers, Teratogens, Marine organism

UNIT V
PREPARED BY
MRS. MEGHA S SHAH
ASSISTANT PROFESSOR, DEPARTMENT OF PHARMACOGNOSY
AISSMS COLLEGE OF PHARMACY, PUNE
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Definition
Fibres may be defined as any hair-like raw material directly obtainable from an animal,
vegetable, or mineral source and convertible into nonwoven fabrics such as felt or paper
or, after spinning into yarns, into woven cloth.
A natural fiber may be further defined as an agglomeration of cells in which the diameter
is negligible in comparison with the length.
Although nature abounds in fibrous materials, especially cellulosic types such as cotton,
wood, grains, and straw, only a small number can be used for textile products or other
industrial purposes.
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Apart from economic considerations, the usefulness of a fibres for commercial purposes
is determined by such properties as length, strength, pliability, elasticity, abrasion
resistance, absorbency, and various surface properties.
Most textile fibres are slender, flexible, and relatively strong.
They are elastic in that they stretch when put under tension and then partially or
completely return to their original length when the tension is removed.
Depending on the sources, fibres are classified broadly in two categories. A. Natural
B. Man made
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Natural Fibres
• Collected from the phloem (the “inner bark” or the skin) or
bast surrounding the stem of certain plant i.e., mainly
dicotyledonous plants.
• Bast fibres from stem: E.g: Flax, Hemp, Jute.
• From leaf: E.g: Ananas, Agave, Palm.
• From seed: E.g: Cotton, Soya, Coir.
• From fruit: E.g: Luffa, Coir etc.
• From grass: E.g: Bamboo, Totora etc.
• From wood: E.g: Hard wood, Soft wood etc.
Vegetable
Fibres /
Bast
Fibres/
skin fibres
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• Mostly cellulose elements, extracted from trees, to make materials
including paper.
• forms of wood fibres including ground wood, thermo mechanical pulp
and bleached or unbleached sulfite pulps
Wood
fibres
• from different parts of animals mostly particular proteins.
• e.g. Silkworm silk, catgut, wool; Hair such as cashmere wool,
mohair and angora; Fur such as sheepskin, rabbit, mink
Animal
fibres
• Short mineral fibres are wollastonite, palygorskite
• Long fibres are asbestos group include chrysotile, amosite,
crocidolite, tremolite, anthophyllite & actinolite
Mineral
fibres
• Fibrous proteins. Consist of biologically important
proteins, mutations etc. E.g: Collagen, actin
Biological
fibres

Man made fibers
• Man-made fibres derived from the natural fibres, mainly cellulose, e.g., nylon
and terylene. Nylons are polymers of adipic acid and hexamethyldiamine,
• whereas terylene is a polymer of ethylene glycol and terephthalic acid
Synthetic
fibre
• Made from raw materials with natural long-chain polymer structure and are only
modified and partially degraded by chemical processes.
• They are regenerated cellulose derivatives. E.g: Rayon, bamboo fibre
Semi synthetic
fibre
• based on synthetic chemicals and made from polyamide nylon, polyester,
polyvinyl chloride etc.
Polymer fibre
• ultra-fine fibres used in filtration. In textiles micro fibres are referred as sub-
denier fibres
Micro fibre
• prepared from the metals such as copper, gold or silver and extruded or
deposited from more brittle ones such as nickel, aluminum or iron
Metallic fibre
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Differences between Natural Fibre and Synthetic Fibres
Natural fibres are derived from plants and animals, whereas synthetic fibres are
almost entirely man made.
Fabrics made of natural fibres are generally more comfortable than synthetic ones.
Natural fibres are expensive compared to synthetic fibres.
In synthetic fibres, spinnerets are used to produce the filaments; whereas, in natural
fibres, it is made naturally.
Natural fibres have limited usage when compared to synthetic fibres.
Natural fibres are biodegradable, hence environmental friendly, whereas synthetic
fibres are not.
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Differences between Plant and Animal Fibre
1. On ignition, animal fibres give bad smell; whereas plant fibres do not give smell.
2. Plant fibres are soluble in cuoxam solution (ammoniacal copper oxide), whereas animal
fibres are 50% soluble in alkaline hydroxide solution.
3. Plant fibres form blue colour when reacted with iodine in the presence of dilute sulphuric
acid; whereas animal fibres form permanent yellow stain when reacted with picric acid.
4. Plant fibres react with Molish’s reagent to form violet colour; whereas with Millon’s
reagent, animal fibres give red
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COTTON
Synonyms: Raw cotton, purified cotton, absorbent cotton.
Biological Source: Epidermal trichomes of the seeds of cultivated species of the Gossypium
herbaceum and other species of Gossypium (G. hirsutum, G. barbadense) freed from
impurities, fats and sterilized, belonging to family Malvaceae.
Geographical Source: The plant is a shrub native to tropical and subtropical regions around the
world, including the America, Africa, Australia and India.
Gossypium hirsutum is native to Central America, the Caribbean and southern Florida,
G. barbadense is native to tropical South America,
G. arboreum is native to India and Pakistan.
Gossypium herbaceum is native to africa and arabia
In India, the major production of the plant is in MH, GJ, PB, AP and MP.
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Cultivation, Collection, and Preparation
Cotton is cultivated by means of seed sowing method.
The seeds are sown in rows of about 4–5 ft in distance. Proper fertilizers are provided timely.
The cotton plants are shrubs or small trees that bare fruits (capsules) after flowering. The capsule
consists of three to five seeds and is covered with hairs.
The bolls are collected when ripe, separated from the capsule, dried, and subjected to the ginning
press for processing.
In ginning process, hairs and seeds are put before the roller with a small space, which separates
the trichomes from the seeds.
The short and long hair separated by delinter. Short hairs are known as ‘linters’, which are used in
the manufacturing inferior grade cotton wool, whereas long hairs are used for preparation of cloth.
The seeds remain after the removal of hair is used for the preparation of cotton seed oil and oil
cake for domestic animal feed.

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The raw cotton so obtained is full of impurities like the
colouring matter and fatty material.
It is then subjected to further purification by treating it with
dilute soda ash solution under pressure for about 15 hours. It
is then bleached and washed properly, dried, and packed. The
packed cotton is then sterilized using radiations.
Description
Colour : White, creamy white
Odour : Odourless
Taste : Tasteless
Size: Cotton fibres are 2.5 to 4.5 cm in length and 25-30
micron in diameter.
It is free from pieces of leaves, seed, coat, foreign matter and
dust. It may be slightly off white in colour, if sterilized.

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Standards:
Absorbent cotton wool I.P has following standard.
1. Length of staple: NLT 15 mm
2. Water soluble extractive: NMT 0.5%
3. Sulphated Ash: NMT 0.5%
Chemical Constituents
It consists of 90% of cellulose, 7–8% of moisture, wax,
fat, oil 0.5% and remains of protoplasm.
Purified cotton has almost cellulose and 6-7 % of
moisture.
Uses
Used as a filtering medium and in surgical dressings.
Used as insulating material.
Absorbent cotton absorbs blood, pus, mucus, and
prevents infections in wounds.

Chemical Tests
1.On ignition, cotton burns with a flame, gives very little odour bead, and leaves a small
white ash; distinction from acetate rayon, alginate yarn, wool, silk and nylon.
2.Dried cotton is moistened with N/50 iodine and 80% w/w sulphuric acid is added. A
blue colour is produced; distinction nylon.
3.With ammoniacal copper oxide solution, raw cotton dissolves with ballooning, leaving a
few fragments of cuticle. Absorbent cotton dissolves completely with uniform swelling,
distinction from acetate rayon, jute, wool, and nylon.
4.In cold sulphuric acid (80% w/w) cotton dissolves; distinction from oxidized
cellulose, jute, hemp, and wool.
5.In cold sulphuric acid (60% w/w) cotton, is insoluble; distinction from cellulose
wadding and rayons.
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6.In warm (40°C) hydrochloric acid it is insoluble; distinction from acetate rayon
(also silk, nylon).
7.It is insoluble in 5% potassium hydroxide solution; distinction from oxidized
cellulose, wool and silk.
8.Treat it with cold Shirla stain A for 1 min, wash out. shows shades of blue, Tilac or
purple; distinction from viscose, acetate rayons, alginate yarn, wool, silk, nylon.
9.Treat it with cold Shirla stain C for 5 min and wash out; raw cotton gives a mauve (pale
purple) to reddish brown colour and absorbent cotton a pink one; distinction from flax,
jute, hemp. The Shirla stains may be usefully applied to a small piece of the whole fabric
under investigation to indicate the distribution of more than one type of yarn.
10.Does not give red stain with phloroglucinol and HCl; distinction from jute, hemp kapok.

JUTE
Synonym: Gunny
Biological Source: The jute fibre is collected from the stem bark of the white jute plant
and to a lesser extent tossa jute belongs to species Corchorus i.e. C. capsularis and C.
olitorius. Family: Tiliaceae or Malvaceae.
Geographical source: Jute is cultivated in Bangladesh, Nepal, Myanmar, India, Thailand,
China, Pakistan, Japan, UK, France, Egypt and Spain.
In India, the major production of jute is in West Bengal, Assam and Bihar.
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Production of Fibre:
The unbranched stems of the jute plant are cut and formed a
bundle. This should carry out before flowering stage.
The jute fibre comes from the stem and ribbon (outer skin) of
the jute plant. The fibres are first extracted by retting method
either in pond or any water logged condition.
Retting is a microbial process by which the jute plant is
immersed in water and the fibre is loosened from the woody
core of the jute plant.
This process softens the tissues and breaks the hard pectin
bond between the bast and jute hurd (inner woody fibre stick)
and the process permits the fibres to be separated.
It has been found that jute stems ret most rapidly at 34ºC.

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At this temperature it takes generally 8-12 days for complete
retting.
The retting process consists of bundling jute stems together and
immersing them in slow running water.
After the retting process, stripping begins. In the stripping
process, non-fibrous matter is scraped off, leaving the fibres to
be pulled out from within the stem and washed in clear, running
water.
Then they are hung up or spread on the attached roofs to dry.
After 2-3 days of drying, the fibers are tied into bundles.
The jute fibres are graded according to colour, length and
smoothness of the fibres

Physical Properties
Colour : Golden yellow
Odour : Characteristic
Taste : None
Size : Average length is 1 to 4 m and diameter from 17 to 20 microns
Appearance : Jute is long, soft and shiny
Chemical Constituents: Jute fibres contain mainly cellulose (60%) and hemicellulose
(24%). Apart from that Lignin (10%), fats and wax are also present.
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Chemical test: The middle lamella is highly liquefied and give red colour with
phloroglucinol and HCl. Indicating the presence of lignin.
Uses: Jute is the second most important vegetable fibre after cotton. Jute is used chiefly to
make cloth for wrapping bales of raw cotton and to make sacks and coarse cloth.
The fibres are also woven into curtains, chair, coverings and carpets.
The fibres are used alone or blended with other types of fibres to make twine and rope.
Used in the manufacture of tows, padding splints, filtering, straining medium and coarse
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HEMP
Biological Source: Hemp is the pericyclic fibre obtained from
Cannabis sativa Linn., belonging to family: Cannabinaceae.
Geographical Source:
Hemp is grown at any altitude from Norway to the Equator.
The raw materials are imported from China, Hungary,
America, Germany, Switzerland, Australia, Canada, France,
and Norway.
In India, MP, Himachal Pradesh and Jharkhand are cultivating
hemp for industrial use.
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Preparation of Fibres
Fibres are obtained by subjecting the stalks to retting, drying and crushing. Finally a
shaking process completes separation from the woody portion, and releases the long, fairly
straight fibre, or line.
Recently new technologies such as ultrasound and steam explosion have been developed
for separation of the fibres from the stalk.
Two principal types of fibres in hemp viz. bast/long fibres and hurds/inner short fibre.
Bast fibres are of two types namely, primary bast fibres and secondary bast fibres.
The primary bast fibres are made up of approximately 70% of the fibres and are long.
They contain high cellulose and low lignin. Primary bast fibres are the most valuable part
of the stalk, and are generally considered to be among the strongest plant fibres known.
Secondary bast fibres are made up of the remaining 30% of the bast fibres and are

Organoleptic characteristic
Colour : Yellowish, greenish, or a dark brown or gray
Odour : Odourless
Taste : Tasteless
Appearance : Made of individual cylindrical cells with an irregular surface, they are longer and
less flexible than flax.
Size : Strand is about 5.8 feet (1.8 metre) long and blunt rounded ends and the diameter ranges
from 16-50 micron.
Chemical Constituents: Hemp fibres mainly contain 77% cellulose and 10% hemicellulose. Apart
from that lignin, fat, wax and pectin are also present.
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Uses:
Natural fibres from the hemp stalk are extremely durable and can be used in the
production of textiles, clothing, canvas, rope, cordage, archival grade paper, paper, and
construction materials.
Long hemp fibres are used in clothing, home furnishing textiles and floor coverings,
whereas short fibres are used in making insulation products, fibreboard and erosion
control mats.
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HALLUCINOGENS
Hallucinogens are psychoactive agents. They cause hallucinations, perceptual anomalies
and other substantial subjective changes in thoughts, emotion, and consciousness.
All hallucinogens contain nitrogen and are classified as alkaloids.
They are found in the various parts of the plant such as roots, leaves, seeds, bark and/or
flowers parts like many hallucinogens contain chemical structures similar to those of
natural neurotransmitters.
Affect neural circuits in the brain involving the neurotransmitter serotonin, and dissociative
drugs cause their effects by disrupting the actions of the glutamate system of the brain.
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The regions of the brain that are affected by hallucinogens control mood, sensory
perception, sleep, hunger, body temperature, sexual behavior and muscle control.
They are administered in various ways namely smoked or snuffed, swallowed fresh or
dried, drunk in decoctions and infusions, absorbed directly through the skin, placed in
wounds or administered as enemas.
They are mainly of three types: psychedelics, dissociatives and deliriants.
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1. Psychedelics: They are class of drugs that trigger psychedelic experiences via
serotonin receptor agonism, causes thought and visual or auditory changes and
altered state of consciousness.
2. Dissociatives: They produce analgesia, amnesia and catalepsy at anesthetic doses.
They achieve their effect through blocking the signals received by the NMDA (N-
methyl-D-aspartate) receptor set. They also have CNS depressant activity.
3. Deliriants: They induce delirium. They are characterized by extreme confusion and
an inability to control one's actions. They are unpopular as recreational drugs due to
the severe and sometimes unpleasant nature of the hallucinations produced.
They block the muscarnic acetylcholine receptors, hence they are known as
anticholinergic drugs.

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Plant name Family Constituents Effects
Catha edulis
(Khat)
Celastraceae Cathine Induce manic behaviours and
hyperactivity, causes loss of appetite.
Coffea Arabica
(Coffee)
Rubiaceae Caffeine Stimulant, temporarily warding off
drowsiness and restoring alertness.
Cannabis sativa
(Marijuana)
Cannabaceae Tetrahydrocan
nabinol
Relaxation and increase in appetite.
Datura stramonium Solanaceae Hyoscine Acts as deliriant and can produce intense
spiritual visions.
Erythroxylum coca Erythroxylaceae Cocaine Stimulant, appetite, suppressant.
Papaver somniferum Papaveraceae Morphine Analgesia, sedation, euphoria
Lophophora
williamsii(Peyote)
Cactaceae Mescaline Hallucinogen.
Nicotiana Tobacum Solanaceae Nicotine Stimulant, relaxant.
Salvia Divinorum
(Salvia)
Lamiaceae Salvinorin-A Induce Hallucination.
Some
of
the
important
herbal
plants
which
are
used
as
a
significant
psychoactive
activity
and
their
effects

TERATOGENS
A teratogen is an agent, which can cause a birth defect via toxic effect on an embryo.
The study of abnormalities of physiological development is known as Teratology.
It results growth retardation, delayed mental development or other congenital disorders
without any structural malformations.
This condition occurs due to drugs used in pregnancy, lack of nutrients such as folic
acid, physical restraint such as Potter syndrome, genetic disorders, alcohol consumption
during pregnancy etc.
There are three different types of possibilities such as: (1) known teratogens in known
teratogenic plants, (2) known teratogenic plants with unidentified teratogens, and (3)
suspected teratogenic plants.
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Biological source Family Constituents Effects
Asparagus
racemosus
(Shatavari)
Liliaceae Shatavarin Cause gross malformations in fetus, can increase
the rate of re-absorption in fetus and may also cause
intrauterine growth.
Conium
maculatum
(Hemlock)
Apiaceae Coniine Coniine Acts directly on the central nervous
system through inhibitory action on nicotinic
acetylcholine receptors.
Leucaena
leucocephala
(River Tamarind)
Fabaceae Mimosine Inhibits DNA synthesis at the level of elongation of
nascent chains by altering deoxyribonucleotide
metabolism.
(Lupine)
Lupinus
mutabilis
Fabaceae Sparteine Cause gross malformations in fetus.
Ruta graveolens
(Ruta)
Rutaceae Arborinine Women for contraception or induced abortion.
Veratrum album
(Veratrum)
Liliaceae Veratramine Act by increasing the permeability of
the sodium channels of nerve cells.
Known Teratogens in known teratogenic plants

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Biological source Family Constituents Effects
Astragalus gummifer
(Tragacanth)
Leguminosae Bassorin Use in spermicidal jelly.
Malus domestica
(orchard apple)
Rosaceae Amygdalin
(from seed)
Large doses can cause adverse
reaction.
Nicotiana tabacum
(Tobacco)
Solanaceae Nicotine Consume during pregnancy,
controls birth.
Trachymene species Apiaceae Alkaloids Roots are a traditional Aboriginal
bushfood.
Prunus amygdalus
(Almond)
Rosaceae Cyanide In large dose causes adverse
reaction.
Known Teratogenic Plants With Unidentified Teratogens

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Suspected Teratogenic Plants
Biological source Family Constituent Effects
Datura stramonium (Datura) Solanaceae Atropine In large dose it causes bradycardia.
Lycopersicon esculentum (Tomato) Solanaceae Solanin It causes nervous disturbances.
Senecio vulgaris (Groundsel) Asteraceae Senecionine It causes irreversible liver damage.
Solanum tuberosum (Potato) Solanaceae Solanin It causes nervous disturbances.
Solanum melongena (Egg plant) Solanaceae Solanin It causes nervous disturbances.
Sorghum arundinaceum (Sorghum) Poaceae Prussic acid In high dose it causes poisoning.

NATURALALLERGENS
Allergens are a type of antigen that produces an abnormally vigorous immune response.
They are inciting agents of the allergy. That means they are the substances that are
capable of sensitizing the body in such a way that an unusual response occurs, in
hypersensitive person.
It may be biological, chemical or synthetic origin. The substances such as pollens,
danders, dust etc. are as natural allergens.
They are protein and glycoprotein in nature. They are mainly of 5 types
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(a) Inhalants: They are air borne substances that are chemicals which cause respiratory diseases,
inflammation in nose, lungs etc.
They are causes by pollen, dust mites, pets and moulds and results Hay fever, Asthma etc.
Symptoms: Sneezing, lacrimation, coughing, itching eyes, nose etc.
(b) Ingestants: These are also known as food allergy. Allergens which are present in food stuff
and swallowed are termed as ingestants.
A food allergy is an immune system that coexists with inhalant allergies. When the foods are
digested and the nutrients are absorbed, substances in food stimulate allergic response.
Foods induce respiratory symptoms by both reaginic and non-reaginic mechanisms.
Most common food allergens ingested by patients are milk, egg, peanut, fish, soy, wheat etc.
Symptoms: Skin rash, migraine, Bronchial asthma, GIT disturbance etc.

(c) Injectants: They are injectable preparations and some insects. They cause allergy in
hypersensitive person.
The natural sources of injectable allergens are produced by the sting of bees, wasps, hornets.
Symptoms: Itching, peeling of skin, Erythema etc.
(d) Contactants: Allergens produce manifestation of hypersensitivity at the site of skin or other
mucous.
Aeroallergens such as the various pollen grains containing oils trichomes from various leaves,
flowers are carried by smoke originating from brush fires, grass fires are also cause for contact
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A number of plant products used as additives in cosmetic preparations are irritants and cause
skin allergy.
Wool fats in cosmetics, soap, soap powder, enzyme detergents, nail polishes, hair dyes are also
major cause of contact dermatitis.
(e) Infectants: Allergy caused by metabolic products of living microorganism in the human body.
The continual presence of certain types of bacteria, molds, protozoas, in the human body being
are responsible for chronic infection.
Sometimes bacterial metabolic wastes are considered to be infectant allergens.
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PRIMARY METABOLITES CARBOHYDRATES
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DEFINATION
They are organic compounds found in the major part of fruits, vegetables, legumes and
cereal grains. They carry out many functions in all living organisms.
These are large biomolecules, consisting of carbon (C), hydrogen (H) and oxygen (O) in
their basic structure.
Chemically, they are simple organic compounds that are aldehydes or ketones with
many hydroxyl groups added on each carbon atom which are not part of either of these
both functional groups.
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The basic formula is Cm(H2O)n where “m” is different from “n”. The hydrogen and
oxygen ratio is 2 : 1, hence they are known as hydrates of carbon.
So, carbohydrate is defined as polyhydroxy aldehydes or polyhydroxy ketones
which give these on hydrolysis and contains at least one chiral carbon atom.
Carbohydrates are produced in green plants by photosynthesis and serve as a major
source of energy in animals.
They also serve as structural components, such as cellulose in plants and chitin in some
animals.
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PROPERTIES
Low molecular weight carbohydrates are crystalline, soluble in water and sweet in taste, for
example, glucose, fructose, sucrose, etc. The high molecular weight carbohydrates (polymers) are
amorphous, tasteless and relatively less soluble in water, for example, starch, cellulose, inulin, etc.
Aldehyde react with alcohol to form acetal, Ketone react with alcohol to form ketals.
Monosaccharide are reducing sugar, whereas disaccharide, polysaccharide are non reducing
sugars.
Monosaccharide reaction with hydrazine to form osazone.
Monosaccharide reduction to form sugar alcohol.
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Carbohydrates act as energy reserves, also stores fuels, and metabolic intermediates.
Ribose and deoxyribose sugars forms the structural frame of the genetic material, RNA and
DNA.
Polysaccharides like cellulose are the structural elements in the cell walls of bacteria and
plants.
Carbohydrates are linked to proteins and lipids that play important roles in cell interactions.
Carbohydrates are organic compounds, they are aldehydes or ketones with many hydroxyl
groups.
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Steroisomerism – Compounds having same structural formula but they differ in spatial
configuration. Example: Glucose has two isomers with respect to penultimate carbon atom.
They are Dglucose and L-glucose.
Optical Activity - It is the rotation of plane polarized light forming (+) glucose and (-) glucose.
Diastereo isomers - It the configurational changes with regard to C2, C3, or C4 in glucose.
Example: Mannose, galactose.
Annomerism - It is the spatial configuration with respect to the first carbon atom in aldoses
and second carbon atom in ketoses.
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CLASSIFICATION
carbohydrates
Simple
sugars
monosaccharides
diasaccharides
trisaccharides
polysaccharid
es
oligosaccharides
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Carbohydrates are of three types namely simple sugar, polysaccharides and
oligosaccharides .
The simplest carbohydrates are the three-carbon sugars, i.e. monosaccharides, and
further cannot be hydrolysed to simple sugars.
Depending on the number of carbon atoms, monosaccharides are further classified as
Bioses are two-carbon compounds, but do not occur in free form in the nature.
Trioses are three-carbon compounds. They are in the form of phosphoric esters.
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Tetroses are four-carbon compounds. Examples: Erythrose, Threose.
Pentoses are five-carbon compounds and occur in plants. They are the products of
polysaccharides such as hemicelluloses, mucilage etc. Examples: Ribose, Arabinose,
Xylose etc.
Hexoses are six-carbon molecules and are abundantly available. On hydrolysis they
produce starch and inulin. Examples: Glucose, Fructose etc.
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Monosaccharides are classified according to three different characteristics: the
placement of carbonyl group, the number of carbon atoms they contain, and their chiral
handedness.
If the carbonyl group is an aldehyde, the monosaccharide is an aldose, and if the
carbonyl group is a ketone, the monosaccharide is a ketose. Aldose or aldo sugar
containing compounds are glucose, galactose, ribose etc., whereas ketose or keto sugar
containing compound is fructose.
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Depending on the chemical nature of the sugar, they are also classified as reducing
and non-reducing sugars.
Reducing sugar: Contain a hemiacetal or hemiketal group. Sugars include glucose,
galactose, fructose, maltose, lactose.
Non-reducing sugar: Hemiacetal groups are absent. Sucrose and all polysaccharides
are present in this group. Heptoses contain seven carbon atoms and so on
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Disaccharides are the compounds in which two monosaccharides are joined together
and these are the simplest polysaccharides. Examples: Sucrose, lactose etc.
Sucrose = Glucose and Fructose; Lactose = Galactose and Glucose; Maltose = Glucose
and Glucose
Trisaccharides are oligosaccharides composed of three monosaccharides with two
glycosidic bonds connecting in between them. Examples: Raffinose (Glucose + Fructose
+ Galactose).
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Polysaccharides: Polysaccharides are polymeric carbohydrates that are composed of
long or branched chains of monosaccharide units bounded together by glycosidic bonds.
They are of two types viz. Structural polysaccharides and Digestible polysaccharides.
Former one are digestible by herbivorous species — cellulose, lignin, dextrans, mannans,
inulin, pentosans, pectic acids, algic acids, agar and chitin. Later one is starch.
Oligosaccharides: Oligosaccharides are saccharide polymers containing a small number
of simple sugars. Some examples are fructo-oligosaccharides (FOS), which are found in
many vegetables.
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Function of carbohydrates
They are used for energy storage and production. Starch and glycogen, respectively in plants and
animals, are stored as carbohydrates from which glucose can be mobilized for energy production.
The presence of carbohydrates is necessary for the normal lipid metabolism.
Glucose is indispensable for the maintenance of the integrity of nervous tissue and red blood
cells.
Two sugars, ribose and deoxyribose, are part of the bearing structure, respectively of the RNA
and DNA and present in the nucleotide structure.
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They take part in detoxifying processes. For example, at hepatic level glucuronic acid,
synthesized from glucose, combines with endogenous substances, as hormones, bilirubin etc.
Carbohydrates are also found linked to many proteins and lipids. Within cells they act as signals
that determine the metabolic fate or intracellular localization of the molecules which are bound.
Two homopolysaccharides, cellulose and chitin, serve as structural elements.
The cellulose in plants is used to manufacture paper, wood for construction, and fabrics.
They exert a protein-saving action.
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Chemical Tests for carbohydrates
Molisch Test: Molisch reagent is mixed with a dilute solution of carbohydrate. The test
reagent dehydrates pentoses to form furfural (top reaction) and dehydrates hexoses to form
5-hydroxymethyl furfural (bottom reaction). The furfurals further react with alpha-
naphthol present in the test reagent to produce a purple product. Pentoses and hexoses
form five-member oxygen containing rings on dehydration. This test is known as
Molisch test and is used to detect carbohydrates in several substances.
Fehling’s Test: Fehling’s solution (containing Cu2+) changes colour from blue to
red/brown in the presence of reducing sugars.
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Benedict’s Test: This test is performed to identify the reducing sugars. Few ml of a sample
solution is placed in a test tube. Two ml of Benedict’s reagent (a solution of sodium citrate
and sodium carbonate mixed with a solution of copper sulfate) is added. The solution is
then heated in a boiling water bath for three minutes. A reddish precipitate will form within
three minutes.
Seliwanoff’s Test: Used to distinguish aldohexoses from ketohexoses. A ketohexose like
fructose form a deep red colour with Seliwanoff’s reagent (a solution of resorcinol in HCl),
while an aldohexose show a light pink colour and takes a longer time to develop the colour.
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Barfoed’s Test: performed to distinguish between mono and disaccharides. Barfoed’s
reagent is copper acetate in acetic acid and is not as reactive as Benedict’s reagent.
A positive reaction may only be a light red precipitate. Monosaccharides produce the red
precipitate in 2 to 3 minutes; disaccharides produce the precipitate in 10 minutes.
Iodine Test for Starch and Other Polysaccharides: Starch is a polysaccharide that can
be easily identified by the iodine test. Cellulose do not form colour complex with
Iodine solution. Starch reacts with iodine solution and forms blue-violet color. Many
glucose in starch trap iodine molecule and form the colour.
54

Bial’s Test: This test is used to distinguish pentoses and hexoses.
Pentoses give a positive test with Bial’s reagent (contains orcinol, HCl and ferric
chloride). In the presence of concentrated HCl, pentoses form a five-membered
ring, known as furfural. A positive test is the formation of a bluish colour within 5
minutes without the formation of a precipitate. Hexoses generally react to form
green, red, or brown products.
55

ACACIA GUM
Synonyms: Acacia gum, Acacia vera, Egyptian thorn, Gummi
africanum, Gum Senegal, Gummae mimosae, Kher, Sudan gum
arabic, Somali gum, Yellow thorn, Indian Gum and Gum Arabic
Biological source: It is a dried gummy exudation obtained from
stem and branches of Acacia arabica, A. senegal.
Family: Leguminosae.
Distribution: The plant is distributed in West Africa, Central Africa,
Europe, Australia, Asia. In India it is collected from Western
Ghats, Punjab, Rajasthan, and Gujarat
56

57
Description of Gum: The gum is round tears and white in colour but sometimes light brown or
cream brown colour also forms, which is inferior quality of gum.
Outer surface of gum is dull and smooth, but dried gum is brittle in nature. The gum is odourless
and mucilaginous in taste.
The gum is soluble in water and forms acid, but is insoluble in alcohol.
Parts Used: Pods, seed and gummy exudation from stem.
Collection and Preparation: After the rainy season, the gum exudes spontaneously from the trunk
and principal branches. Since the flow is small in quantity, it is stimulated by incisions in the
bark. To facilitate the flow, a thin strip, 2 to 3 feet in length and 1 to 3 inches wide is torn off. In
about a fortnight, it thickens and hardens on exposure to the air, in the form of round or oval
tears.
They are white or red, according to whether the species is a white or red gum tree.

Chemical constituents
Acacia consists principally of arabin, which is a
complex mixture of calcium, magnesium and potassium
salts of arabic acid.
Arabic acid is a branched polysaccharide that yields L-
arabinose, D-galactose, D-glucuronic acid and L-
rhmnose on acid hydrolysis.
1, 3-Linked D-galactopyranose units form the backbone
chain of the molecule and the terminal residues of the 1,
6-linked side chains are primarily uronic acids.
Acacia contains 12-15% moisture and enzymes oxidases,
peroxidases and pectinases.
58

Use
Used in pharmaceuticals as a demulcent.
Used topically for healing wounds and shown to inhibit the growth of periodontic bacteria
and the early deposition of plaque.
Used as an emulsifier and a thickening agent in icing, fillings, chewing gum and other
confectionery items.
Reduces cholesterol levels and helps to increase weight loss when taken orally.
Used in diarrhea, irritations, ulcer in the stomach and intestine, bleeding piles, leucorrhoea.
59

Chemical Tests
1. Aqueous solution of gum + Ruthenium red No Pink colour.
2. Aqueous solution of gum + Hydrogen peroxide + Benzidine in alcohol Blue colour
(Due to presence of oxidase enzyme)
3. Aqueous solution of gum + Lead subacetate Gelatinous form
4. Aqueous solution of gum + Ferric chloride No blackish colouration or blackish
precipitate Tannin absent.
5. Aqueous solution of gum + Iodine solution No blue colour (Starch absent).
6. Aqueous solution of gum + dil. HCl Boil Add Fehling’s A and B Red colour
after heating.
60

61
Adulterants and Substituent: Gum Senegal is often used as a substituent for Indian gum.
It is identified as gum Senegal is yellowish white in colour, tears are ovoid, whereas gum
Arabica is whitish in colour, and tears are rounded.
Indian gum is adulterated with gum ghatti obtained from Anogeissus latifolia. It is
identified as it reacts with lead subacetate, but very less precipitate will form and
morphologically, the outer surface of the gum is dull and without fissures. Starch,
sterculia gum and dextrins are also used as adulterants for Indian gum.
Storage: In contact with the moisture the gum becomes hardened and darkened in colour
and this can affect its quality. Hence, it should be stored in clean, cool and dry place. It
does not deteriorate due to long storage under such favorable conditions.

AGAR
Synonym: Agar agar, Japanese Isinglass, Vegetable gelatin.
Botanical Source: It is the dried gelatinous substance obtained by extraction with water from
Gelidium amansii or various species of red algae like Gracilaria and Pterocladia, belonging
to family Gelidaceae
Geographical Source: Japan was the only country producing agar before the World War II,
but it is now produced in several countries like, Japan: Gelidium amnasii and other
Gelidium species, Australia; Gracilaria confervoldes, New Zealand; Pterocladia lucida and
other allied species, Korea, South Africa, United States, Chile, Spain, and Portugal.
62

Agar is a complex heterosaccharide and contains two different polysaccharides known
as agarose and agaropectin.
Agarose is neutral galactose polymer and is responsible for the gel property of agar. It
consists of D-galactose and L-galactose unit.
The structure of agaropectin is not completely known, but it is believed that it consists
of sulphonated polysaccharide in which galactose and uronic acid are partly esterified
with sulphuric acid. Agaropectin is responsible for the viscosity of agar solution.
64

Chemical Tests
Agar sample +sufficient water and boiled. After cooling, stiff jelly is formed.
To agar powder a solution of ruthenium red is added. Red colour is formed indicating
mucilage.
0.2% agar solution in water +Tannic acid but no white precipitated indicate absence of
Gelatin
Sample solution mixed with soda lime but no ammonium smell.
65

66
Aqueous solution of agar is mixed with dil. HCl and then boiled. After that the solution
is divided into two parts. In one part 10% NaOH and Fehling’s solution is added that
gives red colour due to presence of galactose. In another part barium chloride is added
which gives white precipitate due to formation of barium sulphate. This test is absent
in case of starch, acacia gum and tragacanth.
To agar solution an N/20 solution of iodine is added. A deep crimson to brown colour
is obtained (distinctive from acacia gum and tragacanth).
Agar is warmed in a solution of KOH. A canary yellow colour is formed.
Agar responds positively to Fehling’s solution and Molisch reagent.

67
Uses
Agar acts as a solidifying component of bacteriological culture media.
used in canning meat, fish, and poultry; in cosmetics, medicines, and dentistry.
used as thickening agent in ice cream, pastries, desserts, salad dressings, and as a wire-
drawing lubricant.
It functions as food stabilizers. It is also an emulsifying agent and a bulk laxative.
used to treat chronic constipation, as a laxative, suspending agent, a gelating agent for
suppositories, as surgical lubricant, as a tablet excipient, disintegrant, in production of
medicinal encapsulation and ointment and as dental impression mold base.

Substitutes and Adulterants
Some of the common adulterants present in agar are gelatin and Danish agar.
The presence of gelatin can be detected by addition of equal volume of 1% trinitrophenol
and 1% of agar solution; the solution produces turbidity or precipitation.
Danish agar has an ash of 16.5–18.5%, it is formed from rhodophyceae indigenous to the
Denmark costal region.
The Danish agar has a gel strength which is half of its gel strength of Japanese agar.
68

TRAGACANTH
Synonyms: Goat’s thorn, gum dragon, gum tragacanth, hog gum.
Biological Source: It is the air dried gummy exudates, flowing naturally or obtained by incision,
from the stems and branches of Astragalus gummifer and certain other species of Astragalus,
belonging to family : Leguminosae.
Geographical Distribution: The thorny shrubs of tragacanth normally grow at an altitude of 1000-
3000 meter and the primary source is the desert highlands of northern and western part of Iran.
Apart from Iran it is naturally found in various countries, viz., Iraq, Armenia, Syria, Greece and
Turkey. Very few species of Astragalous are located in India, viz., Kumaon, Garhwal & Punjab.
69

Cultivation and Collection
The shrubs of tragacanth are grown altitude of 1000 –3000 m and they are thorny.
The gum is exuding out immediately after injury to plants.
Tragacanth gum is formed as a result of transformation of the cell of pith and
medullary rays into gummy substance.
Incisions are made on various parts of stem.
The fluid which oozes out, is called tragacanth and it is collected after drying.
The gum is in irregular flattened flakes with ribbon like appearance.
It is collected from April to November every year.
70

Description :
Colour : white or pale yellowish Odour : Odourless
Taste : Mucilaginous Size. : 25×12×2 mm in size.
Shape : Thin, flattened ribbon like flakes, more or less curved.
•The gum is horny, translucent, with transverse and longitudinal ridges. Fractures of drug
is short.
•It is partly soluble in water and insoluble in alcohol.
Standards
1.Foreign organic matter : Not more than 1.0%
2.Sulphates Ash : Not more than 4.0%
3.Moisture content : Not more than 15.0%
71

72
Tragacanth gum contains linear chains of galacturonic acid in xylose and with varying
levels of fructose.
Tragacanth contains from 20% to 30% of a water-soluble fraction called tragacanthin
(composed of tragacanthic acid and arabinogalactan).
It also contains from 60% to 70% of a water-insoluble fraction called bassorin.
Tragacanthic acid is composed of D-galacturonic acid, D-xylose, L-fructose, D-
galactose and other sugars.
Tragacanthin is composed of uronic acid and arabinose and dissolves in water to form
a viscous colloidal solution (sol), while bassorin swells to form a thick gel.
Also contains several carbohydrates like L-arabinose, D-galactose, D-Xylose, L-fructose, L-Rhamnose.

Chemical Tests:
Aqueous solution of Tragacanth + Conc. Hydrochloric acid ¾® Boil ¾® No Red
colour forms.
Sample of Tragacanth + Ruthenium red solution ¾®No Pink colour forms
Aqueous solution of Tragacanth + Drops of Ferric chloride ¾® Deep yellow precipitate
froms.
Sample solution + Lead acetate ¾® Heavy white precipitate forms
73

Uses
Used as an emulsifier, binding agent, and demulcent. Orally, tragacanth is used both for
diarrhoea and as a laxative.
Topically, tragacanth is an ingredient in toothpastes, hand lotions, and vaginal creams and
medicinal jellies like spermicidal jelly.
Used as a binding agent for preparations of tablets and pills. Used as adhesives.
In foods, tragacanth is used as stabilizer, thickener and suspending ingredients in salad
dressings, foods, and beverages.
74

Mechanism of Action: Tragacanth contains ingredients that stimulate the movement of
the intestines. It acts as a stabilizer by formation of non-covalent protein–polysaccharide
complexes via interactions by the methoxylated galacturonic acid in the soluble part of
the gum and by the viscosity increase induced by the insoluble bassorin part.
75

Adulterant and Substitutes
Tragacanth gum of lower grades known as hog tragacanth is used in textile industry and in the
manufacture of pickles.
The gum varies from yellowish brown to almost black.
Citral gum obtained from A. strobiliferus is also used as an adulterant.
Karaya gum which is sometimes known as sterculia gum or Indian tragacanth is invariably used
as a substitute for gum tragacanth. (Karaya gum is identified with the ruthenium solution test
with formation of red colour, whereas this test is negative for tragacanth.)
Dextrin, wheat and corn starch etc. are used as substituents for tragacanth gum.
76

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
Geographical Location: Honey is produced in many parts of the world like Africa,
Australia, Newzealand, Asia,Jamaica, California, Chili, Great Britain.
In India it is abundantly produced in forest area of all the states. Maximum honey
production is obtained from Himalayan forest region, Karnataka, W B, MP and AP.
77

Collection and Preparation
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 applying the pressure
to it or allowing it to drain naturally.
78

79
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.

Honey is essentially a concentrated aqueous solution of inverted sugar and contains a
highly complex mixture of other carbohydrates, a variety of enzymes, amino acids, organic
acids, minerals, vitamins, aromatic substances, pigments, waxes, etc.
The main sugars present in honey are fructose (38%) and glucose (31%).
The saccharose content varies in accordance with the state of maturity of the honey, and
the composition of the oligosaccharide fraction is determined by the plants used in the
production process.
81

Honey contains free amino acids in quantities of around 0.1% of the dry product.
Proline is the major amino acid, but other amino acids like arginine, alanine, glutamic
acid, aspartic acid, lysine, glycine and leucine are also present.
The main acid in honey is gluconic acid and smaller quantities are also found of lactic,
citric, succinic, formic, malic, acetic, maleic and oxalic acids.
82

83
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.
3. Limit Tests: The limit tests of chloride, sulphate and ash (0.5%) are compared with the
pharmacopoeial specifications.

Uses
Used as mild laxative, bactericidal, sedative, antiseptic and alkaline characters.
Used for cold, cough, fever, sore eye and throat, tongue and duodenal ulcers, liver disorders,
constipation, diarrhoea, kidney and other urinary disorders, pulmonary tuberculosis, marasmus,
rickets, scurvy and insomnia.
It is applied as a remedy on open wounds after surgery. It prevents infection and promotes
healing.
Honey works quicker than many antibiotics because it is easily absorbed into the blood stream.
84

85
It is also useful in healing of carbuncles, chaps, scalds, whitlows and skin inflammation; as
vermicide; locally as an excipient, in the treatment of aphthae and other infection of the oral
mucous membrane.
Honey, mixed with onion juice, is a good remedy for arteriosclerosis in brain. Diet rich in
honey is recommended for infants, convalescents, diabetic patients and invalids.
Honey is an important ingredient of certain lotions, cosmetics, soaps, creams, balms, toilet
waters and inhalations.
used as a medium in preservation of cornea.
Potent antibacterial peptides (apidaecins and abaecin) have been isolated and characterized
in the honeybee (Apis mellifera) itself and a new potent antibacterial protein named royalisin
has been found in the royal jelly of the honeybee.

Adulterant and Substitutes
Due to the relatively high price of pure honey, it is invariably adulterated ether with
(a) Honey with Sugar Solution (Sugar + Water): To detect the adulterant, Pure honey is
always in a semi solid state. If adulterated honey is poured in water it will dissolve
immediately.
(b) Honey with Cane Sugar
(c) Honey with Invert Sugar: It can be checked with the help of Fiehe’s test. Sample of
(d) Honey with Glucose: This can be identified by iodine test.
(e) Honey with Commercial Sugar: This can be identified by Aniline chloride test.
(f) Honey with Starch or Flour
86

PRIMARY METABOLITES
PROTEINS AND ENZYMES
87

Definition
The term protein, derived from the Greek proteios, meaning first, are a class of organic
compounds that are present in and vital to every living cell.
They required for the structure, function, and regulation of the body’s cells, tissues, and
organs.
Proteins are large biochemical compounds (carbon, hydrogen, oxygen, and nitrogen)
consisting of one or more polypeptides (amino acid residue) typically folded into a
globular or fibrous form in a biologically functional way.
88

A polypeptide is a single linear polymer
chain of amino acids bonded together
by peptide bonds between the carboxyl
and amino groups of adjacent amino
acid residues i.e. an amine group
(NH2), a carboxylic acid group (R–
C=O–OH) and a side-chain (usually
denoted as R).
89

General Chemical Tests for Proteins
Biuret Reaction: Sample solution + 10% sodium hydroxide + 0.1% copper sulphate solution.
violet or pink colour.
Compounds with two or more peptide bonds give a violet colour with alkaline copper sulphate
solution. Proteins in the alkaline environment reduce Cu2+ to Cu+, which forms a coordination
complex with proteins, leading to a blue to light violet colour change.
Xanthoproteic Reaction: Sample solution + concentrated nitric acid Δ Then 40% sodium
hydroxide is added slowly yellow colour of solution turns to deep orange colour. The yellow
colour is due to the nitro derivatives of the aromatic amino acids present in the protein. The
sodium salts of nitro derivatives are orange in colour
90

Sulphur Test: Sample solution + 40% NaOH + few drops of 2% lead acetate solution
Δ forms black precipitate after cooling.
Sakaguchi Reaction: Sample solution + 0.02% alpha naphthol solution + 10% sodium
hydroxide + few drops of alkaline hypobromide solution. intense red colour.
Ninhydrin Test: Sample solution + 0.1% freshly prepared Ninhydrin solution Δ
violet or purple colour.
91

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
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.
92

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.
93

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.
94

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 color,
finished product is subjected
to bleaching by sulphur
dioxide.
Bleaching affords a light
coloured gelatin.
95

Characteristics
Colour: Colourless or slightly yellow, transparent,
brittle,
Odour: Odourless,
Taste: tasteless sheet, flakes or course granular
powder.
In water it swells and absorbs 5–10 times its weight
of water to form a gel in solutions below 35-
Insoluble : cold water and organic
solvents,
Soluble: Hot water, glycerol, acetic acid;
amphoteric.
In dry condition it is stable in air, but
when moist or in solution, it is
attacked by bacteria.
96

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)
with the help of a Bloom gelometer.
Jelly strength is used in the preparation of
suppositories and pessaries.
Commercially two types of gelatin A and B
Type A has an isoelectric point between pH 7
and 9. (incompatible with anionic compounds
such as Acacia, Agar and Tragacanth.)
Type B has an isoelectric point between 4.7 and
5, (used with anionic mixtures.)
Gelatin is coloured with a certified colour for
manufacturing capsules or for coating of tablets.
It may contain various additives.
97

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.
98

Chemical Tests
1. Biuret reaction: Alkaline solution of a protein +dilute solution of copper sulphate
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 + concentrated nitric acid Δ a yellow colour
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) + Gelatin white precipitate, which turns red on heating.
99

4. Ninhydrin test: To an aqueous solution of a protein + alcoholic solution of
ninhydrin Δ Red to violet colour.
5. On heating gelatin (1 g) with soda lime, smell of ammonia is produced.
6. A solution of gelatin + tannic acid (10%) white buff coloured precipitate
7. Picric acid + gelatin yellow precipitate.
100

Uses
To prepare pastilles, pastes, suppositories, capsules, pill-coatings, gelatin sponge;
As suspending agent, tablet binder, coating agent, as stabilizer, thickener and texturizer
in food;
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.
101

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.
102

CASEIN
Casein is related to phosphoprotein.
These proteins are commonly found in mammalian milk, making up 80% of the proteins
in cow milk and between 20% and 45% of the proteins in human milk.
casein is structured in voluminous globules.
These globules are mainly responsible for the white colour of the milk.
According to various species, the casein amount within the total proteins of the milk
varies.
103

104
Isolation
1. Specified amount of milk is kept in the flask and heated at 40°C in a water bath.
2. Few drops of glacial acetic acid are added and stirred.
3. The resultant mixture is filtered through filter paper held in a funnel and most of the
liquid is gently squeezed out.
4. Casein and fat are removed from the cheesecloth, the solid is placed into a beaker
and few ml of 95% ethanol is added.
5. Then it is stirred well to break up the product. The liquid is poured off and few ml of 1 : 1
ether-ethanol mixture is added to the solid.
6. It is stirred well and filtered through filer paper.
7. Solid is scraped into a weighed filter paper and then dried in the air.
8. The casein content is then calculated as follows: % Casein = gm of casein × 100
gm of milk
Normal Range is 3-5%.

Properties
1. It is purified powder and yellow in colour.
2. It is found in milk as a suspension of
particles called “casein micelles”.
3. It is relatively hydrophobic.
4. It is poorly soluble in water and insoluble
in neutral salt solution.
Casein does not coagulate on heating.
5. The caseins in the micelles are held together by
calcium ions and hydrophobic interactions.
6. Isoelectric point of casein is 4.6.
7. It is readily dispersible in dilute alkalis and in salt
solutions such as sodium oxalate and sodium acetate.
8. Melting point: 280°C.
It is precipitated by acids and by a proteolytic
enzyme (rennet)
105

The principal casein fractions are alpha (s1) and alpha (s2)-caseins, β-casein and κ-
casein.
The distinguishing property of all casein is their low solubility at pH 4.6. The
common compositional factor is that caseins are conjugated proteins, most with
phosphate group(s) esterified to serine residues.
These phosphate groups are important to the structure of the casein micelle.
106

107
Calcium binding by the individual caseins is proportional to the phosphate content.
Within the group of caseins, there are several distinguishing features based on their
charge distribution and sensitivity to calcium precipitation:
Alpha (s1)-casein: (m.w 23,000; 199 residues, 17 proline residues)
Two hydrophobia regions, containing all the proline residues, separated by a polar
region, which contains all but one of eight phosphate groups. It can be precipitated at
very low levels of calcium.
Alpha (s2)-casein: (m.w 25,000; 207 residues, 10 prolines)
Concentrated negative charges near N-terminus and positive charges near C-terminus.
It can also be precipitated at very low levels of calcium.

β-casein: (m.w 24,000; 209 residues, 35 prolines)
Highly charged N-terminal region and a
hydrophobia C-terminal region. Very
amphiphilic protein acts like a detergent
molecule.
Self association is temperature-dependent; will
form a large polymer at 20°C but not at 4°C.
Less sensitive to calcium precipitation.
κ-casein: (m.w 19,000; 169 residues, 20 prolines)
Very resistant to calcium precipitation,
stabilizing other caseins. Rennet cleavage at the
Phe l05 – Met l06 bond eliminates the stabilizing
ability, leaving a hydrophobia portion, para- κ-
casein and a hydrophilic portion called κ-casein
glycomacropeptide (GMP), or more accurately,
caseinomacropeptide (CMP).
108

Uses
Casein is the major component of cheese.
Used as a food additive, binder for safety matches.
As a food source, casein supplies amino acids, carbohydrates and the two inorganic elements
calcium and phosphorus.
Derivatives of Casein are used in tooth remineralization products to stabilize amorphous calcium
phosphate.
Casein peptides are used for high blood pressure, high cholesterol, anxiety, fatigue, epilepsy,
intestinal disorders, cancer prevention and stress reduction.
109

PROTEOLYTIC ENZYMES
Enzymes are proteins that catalyze biochemical functions.
They are required for various physiological processes.
Proteolytic enzymes are also known as protease that digest proteins,
i.e. breakdown of long chain of protein molecules into shorter peptides and their
components such as amino acids.
They act as digestive aids, blood cleansers, rebalance immune system and reduce
oedema in inflamed region.
110

PAPAIN
Synonyms: Papayotin, vegetable pepsin, tromasin, arbuz.
Biological Source: Papain is the dried and purified latex of the green fruits and leaves of Carica
papaya L., belonging to family Caricaceae.
Geographical distribution: The plant is cultivated in Sri Lanka, Tanzania, Hawai, and Florida.
The plant is 5–6 m in height bearing fruits of about 30 cm length and a weight up to 5 kg.
The epicarp adheres to the orange-coloured, fleshy sarcocarp,
which surrounds the central cavity.
This cavity contains a mass of nearly black seeds.
111

Preparation
It is distributed throughout the plant, but mostly concentrated in the latex of the fruit.
The latex is obtained by making two to four longitudinal incisions, about 1/8 inch deep,
on the surface on four sides of nearly mature but green fruits while still on the tree.
The incisions are made early in the morning, at intervals of three to seven days. The
latex flows freely for a few seconds but soon coagulates.
The exudate is collected in nonmetallic containers. The latex is dried as soon as
possible after collection.
112

Rapid drying or exposure to sun or higher temperature above 38°C produce dark colour
product with weak in proteolytic activity.
The use of artificial heat yields the better grade of crude papain.
The final product should be creamy white and friable.
It is sealed in air-tight containers to prevent loss of activity. If 10% common salt or 1%
solution of formaldehyde is added before drying, the product retains its activity for many
months.
Fully grown fruits give more latex of high enzyme potency than smaller or immature
fruits. The yield of Papain varies from 20 to 250 g per tree.
The yield of commercial Papain from latex is about 20%.
113

114
Characteristics
Colour: white or greyish-white,
Nature: slightly hygroscopic powder.
Solubility: Insoluble in water and glycerol.
Digest about 35 times its weight of lean meat.
Best grades render digestion of 200–300 times their
weight of coagulated egg albumin in alkaline
media.
Activators: thiols and reducing moieties (cysteine,
thiosulphate, and glutathione.), activated by
reduction (HCN and H2S)
Properties: Papain is atypical protein digesting
enzyme with isoelectric point, 212 amino
acids, M.W 23,000 daltons, resistant to heat.
Stability: Crystalline papain is most stable in the
pH range 5–7 and is rapidly destroyed at 30°C
below pH 2.5 and above pH 12. Best pH 5.0
Temperature range: 60–90°C is favourable ,
65°C the optimum point.
Inhibitors: by metal ions, oxidants and reagents
which react with thiols and endopeptidase.
inactivated by oxidation (H2O2, iodoacetate).

115
Papain contains several enzymes such as proteolytic enzymes peptidase I capable of
converting proteins into dipeptides and polypeptides, rennin-like enzyme, clotting
enzyme similar to pectase and an enzyme having a feeble activity on fats.
The enzymes, papain, papayaproteinase, and chymopapain, have been isolated in
crystalline form from the latex. Papain contains 15.5% nitrogen and 1.2% sulphur.
The leaves possess dehydrocarpaines I and II, fatty acids, carpaine, pseudocarpaine
and carotenoids.
The fruits yield lauric, myristoleic, palmitoleic and arachidic acids, malonated benzyl-
p-o-glucosides, 2-phenyl ethyl glucoside, and 4-hydroxy-phenyl-2-ethyl glucoside.

Uses
Used to prevent adhesions; in infected wounds; internally as protein digestant,
as anathematic (nematode), to relieve the symptoms of episiotomy (incision of vulva),
in meat industry for tenderizing beef, for treatment of dyspepsia,
intestinal and gastric disorders, and diphtheria, for dissolving diphtheria membrane; in
surgery to reduce incidence of blood clots where thromboplasma is undesirable
for local treatment of buccal, pharyngeal, and laryngeal disorders.
116

117
 Used in digestive mixtures, liver tonics, for reducing enlarged tonsils, in prevention
of postoperative adhesions, Curbuncles, and eschar burns.
 An allergic agent causing Severe paroxysmal cough, vasomotor rhinitis and dyspnea.
 Powerful poison when injected i.v
 In Industry it is used in the manufacture of proteolytic preparations of meat, lever,
and casein, with dilute alcohol and Lactic acid as meat tenderizer, as a substitute for
rennet in cheese manufacture, in brewing industry for making chillproof Bear, for
degumming natural milk, in preparation of Tooth pastes and cosmetics,
 in tanning industry for bathing Skin and hides, and as an ingredient in cleansing
solutions and For soft contact lenses.

Test
1. Papain is reacted with a gelatin solution at 80°C in the presence of an activating
cysteine chloral hydrate solution for an hour. The solution is cooled to 4°C for long
time. The treated solution must not regel in comparison to a blank solution under
identical conditions.
Adulteration
Commercial papain is often adulterated with arrowroot starch, dried milk of cactus, gutta
percha, rice flour, and pepsin.
118

BROMELIN
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,
Thailand,U.S, Brazil, Philippines, Mexico, Hawaii and Taiwan.
119

120
Cultivation, Collection, and Preparation
Bromelin is found in pineapple fruit juice and stem.
Pineapple 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 ammonium sulphide.

Characteristics
Optimum pH: 5.0–8.0. In solution pH
below 3.0 and above 9.5 inactivates the
enzyme.
Optimum temperature: 50 -60°C, still
it is effective between 20 and 65°C too.
Moisture content: NMT 6%.
Colour: light brown coloured powder.
It is not a single substance, but collection of
enzymes and other compounds.
Mixture of sulphur-containing protein-digesting
enzymes, called proteolytic enzymes or proteases.
Several other substances in smaller quantities,
including peroxidase, acid phosphatase, protease
inhibitors, and calcium.
121

Uses
Effective fibrinolytic agent
Inhibits platelet aggregation and seems to have both direct as well as indirect actions
involving other enzyme systems in exerting its anti-inflammatory effect.
Antibiotic potentiation is one of the primary uses of bromelain in several foreign countries; 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.
122

123
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);
Useful in the prevention and treatment of thrombosis and thrombophlebitis. If
administered for prolonged time periods, bromelain also exerts an antihypertensive effect
in experimental animals.
Useful in the treatment of AIDS to stop the spread of HIV.

STREPTOKINASE
Synonym: Estreptokinase,
plasminokinase.
Biological Source: Estreptokinase,
plasminokinase is a purified bacterial
protein produced from the strains of
group C Beta haemolytic streptococci.
Family: Streptococcaceae
124

Characteristics
Streptokinase is a bacterial protein with half-life of
23 minutes and its anisolylated plasminogen activator
complex (APSAC) has a higher half-life of six hours.
Colour: white powder
Solubility: Soluble in water
Optimum pH: 7-8. (7.5)
Isoelectric H : 4.7
Streptokinase is the purified
bacterial protein (extracellur
enzyme) containing single
chain polypeptide,with about
484/414 amino-acid residues
and molar mass is 47 kDa.
125

127
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.

SERRATIOPEPTIDASE
Synonym: Serrapeptase, serratiopeptidase.
Biological Source: Serratiopeptidase is a proteolytic enzyme isolated from nonpathogenic
enterobacteria Serratia E 15. Family: Enterobacteriaceae It is also produced by the larval
form of the silk moth.
Preparation: Serratiopeptidase is produced by fermentation technology by using nonpathogenic
enterobacteria species such as Serratia E 15.
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.
128

Properties
It has high degree of substrate specificity.
The molecular weight of Serrapeptase ranges from about 45 kDa – 60 kDa.
It is a metalloprotease and contains three zinc atoms as ligands and one active site.
The presence of zinc atom is essential and also enhances the proteolytic activity of
Serrapeptase.
The gene encoding Serrapeptase reveals that it is made up of 470 amino acids.
129

The amino acid sequence is free of Sulphur containing amino acids, cysteine and
methionine.
The maximum enzyme activity of Serrapeptase is observed at pH 9.0 and at a temperature
of 40°C. Destroyed by acid in stomach
It is degraded or inactivated completely at a temperature of 55°C.
It is an active enzyme that binds to the alpha-2 macroglobulin in biological fluids and in
blood, it binds in the ratio of 1 : 1 and this binding helps to mask its antigenicity.
The doses usually range from 10 mg to 60 mg per day.
130

131
Uses
Serratiopeptidase is the most widely prescribed anti-inflammatory enzyme in developed
countries and also in India.
Eliminates inflammatory oedema and swelling, accelerate liquefaction of pus and
sputum, and enhance the action of antibodies.
Used as a fast wound healing agent.
Proving to be a superior alternative to the NSAID drugs traditionally used to treat
rheumatoid arthritis and osteoarthritis.
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 of urokinase by recombinant technology
Urokinase is a fibrinolytic enzyme produced by recombinant DNA using genetically
manipulated E. coli cells. It is produced firstly as prourokinase q.v. and then converted
to active form by plasmin or kallikrein.
132

Isolation of urokinase from human urine
Human Urine
Sod. Benzoate
and Dil HCL
is added at pH
4.5
Precipitate is
formed, which
is filtered and
benzoic acid
acid is added .
Acetone is
added to
dissolve
benzoic acid
and urokinase
precipitated
out.
Washed with
acetone and
ether and
dried
Isolation by ion
exchange
chromatography using
phosphate buffer
Lyophilized to
get powdered
enezyme
133

Characters: They are plasminogen activator .
Urokinase is a 411-residue protein, consisting of the domains: the serine protease domain,
the kringle domain and the growth factor domain.
Urokinase is synthesized as a zymogen form and is activated by proteolytic cleavage between
Lys 158 and Ile 159. The two resulting chains are kept together by a disulphide bond.
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.
134

Urokinase enzymes are serine proteases that occur as a single low molecular weight (33
kDa) and double, high molecular weight (54 kDa) polypeptide chain forms.
They differ in molecular weight considerably.
A single chain is produced by recombinant DNA technique and is known as SCUPA
Uses: It is thrombolytic agent, used in treatment of severe or massive deep venous thrombosis,
pulmonary embolism, myocardial infarction and dialysis cannulas.
Used intrapleurally to improve the drainage of complicated pleural effusions and empyemas.
135

PEPSIN
Biological Source: Pepsin is the principal proteolytic enzyme of vertebrate gastric juice.
Properties: Molecular weight of Pepsin: 34.5 kDa
It is an endopeptidase enzyme.
It is buff coloured or white coloured amorphous powder.
It has little acidic or saline taste with slight meaty odour.
It is soluble in water but insoluble in alcohol, ether and chloroform.
It breaks down proteins into peptones and proteases.
136

Pepsin is a monomeric, two domain, mainly beta protein with a high percentage of acidic
residues.
Produced in the stomach and is one of the main digestive enzymes in the digestive system.
It has three-dimensional structure, of which one or more polypeptide chains twist and fold,
bringing together a small number of amino acids to form the active site.
It is an aspartic protease, using a catalytic aspartate in its active site.
It is efficient in cleaving peptide bonds between hydrophobic and aromatic amino acids
such as phenylalanine, tryptophan and tyrosine.
137

Pepsinogen is the proenzyme of pepsin.
Pepsin is most active in acidic environments between 37 °C and 42 °C.
Its primary site of synthesis and activity is in the stomach (pH 1.5 to 2).
Pepsin exhibits maximal activity at pH 2.0 and is inactive at pH 6.5.
Optimal pH: 1.0-4.0 Isoelectric Point: 1.0
There are four reported pepsin proteins: pepsin A, pepsin B (parapepsin I), pepsin C
(gastricsin), and pepsin D (an unphosphorylated version of pepsin A).
138

139
Preparation
Mucous membrane is separated from the stomach either
by the process of stripping or it is scrapped off, and
placed in acidified water for autolysis at 37°C for 2 hours.
After autolysis it consist of both pepsin and peptone.
Filtered and sodium or ammonium salts are added to the
liquid till it is half saturated.
At this point only the pepsin separates out, and the
peptone remains in the solution.
The precipitates are collected and subjected to dialysis
for the separation of salts.
Remaining amount of pepsin if any in the aqueous
solution is precipitated by the addition of alcohol into it.
The pepsin is collected and dried at low temperature.

Activator: Pepsinogen.
Inhibitor: Pepstatin is a low molecular weight compound and potent inhibitor specific
for acid proteases with a Ki value of about 1−10 M for pepsin. Sucralfate also inhibits
pepsin activity.
Specificity: Pepsin has broad specificity for peptides containing linkages with aromatic
or carboxylic L-amino acids.
It preferentially cleaves C-terminal to Phe and Leu and to a lesser extent Glu linkages. The
enzyme does not cleave at Val, Ala, or Gly.
140

Uses
Digestion of antibodies.
Preparation of collagen for cosmeceutical purposes.
Assessment of digestibility of proteins in food chemistry.
Subculture of viable mammary epithelial cells.
Storage: Pepsins should be stored at very low temperatures (between −80°C and −20°C)
to prevent autolysis.
141

PRIMARY METABOLITES
LIPIDS(WAXES, FATS, FIXED OILS)
142

Castor oil
Synonyms: Castor bean oil, castor oil seed,
oleum ricini, ricinus oil, oil of palma christi,
cold-drawn castor oil.
Biological Source: Castor oil is the fixed oil
obtained by cold expression of the seeds of
Ricinus communis Linn., belonging to family
Euphorbiaceae.
Geographical Source: It is mainly found in
India, Brazil, America, China, Thailand; in
India it is cultivated in Gujarat, Andhra
Pradesh, and Karnataka.
143

Preparation
Castor oil is obtained from castor seeds. The oil is obtained by two ways; either after the removal of
the seed coat or with the seed coat.
Seed coats are removed by crushing the seeds under the grooved rollers and then they are subjected
to a current of air to blow the testas.
The kernels are fed in oil expellers and at room temperature they are expressed with 1 to 2 tons
pressure per square inch till about 30% oil is obtained.
The oil is filtered, steamed 80–100°C to facilitate the coagulation and precipitation of poisonous
principle ricin, proteins and enzyme lipase present in it.
144

145
Oil is then filtered and this oil with 1% acidity is used for medical purpose.
The oil cake which remains contains of ricin, lipase and about 20% oil.
The cake is grounded, steamed to 40° to 80°C, and a pressure of 3 tons pressure per sq. inch is
applied.
This yields the second quality of oil with 5% acidity and is used for industrial purpose.
The residual cake which remains after the expression of the second quality oil still contains
about 8 to 10% oil.
This oil is obtained by subjecting it to extraction in soxhlet with lipid solvents. This oil
obtained is also used in industry.
The residual cake is used as manure and not fed to animal due to the presence of ricin. The
cake is also used for the production of lipase.

Physical Properties of Oil
Type : Fixed oil.
Colour : Pale yellow to colourless.
Odour : Soft and faint.
Taste : Highly unpleasant.
Solubility : Soluble in alcohol, organic
solvents like benzene, chloroform but
insoluble in other mineral oils.
Physical Standards
Density (Wt/ml) : 0.945 - 0.965 g/cm3
Acid value : Not more than 2
Flash point : 229.4°C
Refractive index : 1.473 - 1.477
Saponification value : 176 - 187
Iodine value : 81 - 90
Acetyl value : Not less than 142
Viscosity : 6.8 poises
Optical rotation : Between +3.5° and + 6.0°
146

Chemical Constituents:
The oil is made up of triglycerides of 91-95%
ricinoleic acid, 4-5% linoleic acid, and 1-2%
palmitic and stearic acids.
The viscosity mainly depends on the
presence of ricinoleic acid (95%).
Apart from that Oleic acid (6%), Linolenic
acid (5%), stearic acid and palmitic acid are
also present.
147
Ricinoleic acid

148
Chemical Tests
1. Castor oil is missible with half its volume of light petroleum ether at 55°C.
2. An equal proportion of oil and ethanol gives clear liquid even after storage of 3-4 hours;
there will be no change in clarity.
Uses
(a) Industrial Applications:
In the textile industry, castor oil is used for moisturizing and removal of grease in fabrics,
and for the manufacturing of waterproof fabrics.
In the steel industry, it is used in cutting oils and lubricants for steel lamination at high
temperatures and it is also used in other liquids that are necessary for steel work.

The automotive industry uses castor oil for the production of high performance motor
oil and braking fluids, in the production of fluids for hydraulic devices, artificial
leather, varnish, paint, linoleum, insulators, powder, fatty acids, enzymes as a
moisturizer for stationary. here will be no change in clarity.
(b) Medicinal Uses:
Castor oil, seeds, leaves and roots have numerous medicinal importance.
149

Mainly the oil is used as laxative. Besides, these are utilized in peritonitis, diarrhea,
dysentery, lumbago, constipation, piles, paralysis, sciatica, boils, asthma, dropsy,
leprosy, arthritis, amenorrhoea backache, rheumatoid arthritis, ano-rectal problems,
burning feet, period pain, sores, boils, chest, back or abdomen pain, headache, broken
tooth, joint pains, pelvis pain, uterine pain, dermatitis, eczema, lactation, nodules in
breasts etc.
Ricinoleic acid, the active constituent of the oil, is effective against growth of various
species of viruses, bacteria, yeasts and molds.
150

Chaulmoogra oil
Synonyms: Hydnocarpus oil; gynocardia oil.
Biological Source: Chaulmoogra oil is the fixed oil obtained by
cold expression from ripe seeds of Taraktogenos kurzii King,
(syn. Hydnocarpus kurzii (King) Warb.), H. wightiana Blume,
H. anthelminticta Pierre, H. 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.
151

Extraction of Oil
Generally the oil is extracted from the ripe
seed by cold expression method through
hydraulic press.
Fruits are peeled by knife and seeds are
washed in water and dried in sun.
Seeds are decorticated by millet, hand
hammers or by decoricator. Kernels
yield 43 – 48% oil in ghani. The kernel is also
crushed in expeller and rotary.
Extracted oil is stored in zinc barrels and
exported.
Physical properties:
Colour : yellow
Odour : characteristic
Taste : acrid
Solubility : soluble in organic solvents
like chloroform, benzene, Alcohol etc.
State : white solid below 25°c and soft.
Refractive index, at 40°c : 1.472 - 1.476
Sap value : 198 - 204
Acid value : not more than 10
Density : 0.935 - 0.960
Specific rotation : +46° to +60°
152

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.
The seeds of H. Wightiana contain a
flavonolignan Hydnocarpin;
isohydnocarpin, methoxy
hydnocarpin, apigenin, Luteolin,
chrysoeriol, hydnowightin,
epivolkenin, and cyclopentenoid
cyanohydrin glycosides.
153

Substitution: The original oil is
sometimes substituted with the oil
obtained from
Hydnocarpus wightiana, H. alpine
which are abundantly available in
Eastern and Southern
parts of India.
Uses
The oil is useful in leprosy and many
other skin diseases like psoriasis etc.
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
154

SHARK LIVER OIL
Synonyms: Oleum selachoide.
Biological Source: Shark liver oil is the fixed oil obtained from the fresh and healthy
livers of shark fish Hypoprion brevirostris, belonging to family Carcharhinidae.
Geographical Source: Shark is found on seacoasts of many
European countries and in India
in Tamil Nadu, Maharashtra, and Kerala.
155

156
Preparation
Characteristics
 Livers are removed from the fish, cleaned
thoroughly, freed from fatly substances, and
attached tissues like gallbladders.
 Then the livers are heated in water at about
80°C.
 The oil exudes, floats on the top, and is
separated, washed and water is removed. The
dehydrated oil is cooled to separate stearin.
 The suspended materials are removed by
centrifugation.
 The oil is supplemented with vitamins A and
D in desired amount.
Colour: pale yellow to brownish yellow,
viscous liquid
Odour: fishy odour Taste: bland taste
Solubility: Insoluble in water, sparingly
soluble in alcohol and freely miscible in
nonpolar solvents such as petroleum
ether,chloroform, and benzene.
acid value: 2
saponification value: 150–200,
iodine value: 160–350.

The active principle of Shark liver
oil is vitamin A which varies from
15,000 to 30,000 I.U. per g of the
oil.
It contains glycerides of saturated
and unsaturated fatty acids.
157

Chemical Tests
1. Shark liver oil (1 drop) in chloroform
(1 ml) sulphuric acid violet colour
change to purple or brown is formed due
to the presence of vitamin A.
2. Shark liver oil (1 ml) is dissolved in
chloroform saturated solution of
antimony trichloride in chloroform
blue colour
is formed due to the presence of vitamin
A.
Uses
Shark liver oil is used to treat
xerophthalmia (abnormal dryness of the
surface of conjunctiva) occurring due to
deficiency of vitamin A.
The oil is nutritive and used as a tonic.
Marketed products
It is one of the ingredients of the
preparation known as Shark liver oil
softgels (now foods).
158

COD LIVER OIL
Biological Source: It is processed from
fresh liver of cod fish, Gadus morrhua
and other species of Gadus, belonging to
family Gadidae.
Geographical Source: It is mainly found
in Scotland, Norway, Germany,
Iceland, and Denmark.
Characteristics
Colour: pale yellow in colour
Odour and taste: fishy odour taste
Solubility: slightly soluble in alcohol and
fully soluble in chloroform, ether, carbon
disulphide and petroleum ether.
Specific gravity: 0.922–0.929,
Refractive index: 1.475–1.4745,
Acid value: less than 2,
Iodine value: 155–173.
159

160
Preparation
The liver is cleaned and minced into small pieces and heated to 80°C in a vat by
admitting steam for half an hour.
The enzyme lipase is destroyed at temperature above 70°C.
The oil is removed and put in tin drums which are encased
with wooden barrels.
The barrels are kept inside the snow and the oil is cooled to -2 to -5°C, the slow
cooling process precipitates the palmitin, which is separated by filtration.
The oil obtained is medicinal oil. The residual cake formed after the medicinal oil is
subjected to heating at higher temperature to obtain oil with inferior quality and brown
colour.

C.C
Contains glycerides esters of saturated
acids of linoleic, oleic, myristic, gadoleic,
palmitic, and other acids.
The oil has vitamin A and vitamin D.
Also contains about 1% unsaponifiable
matter; like cholesterol, fatty alcohol,
squalene, α-glyceryl esters, etc.
Uses
Oil is used as source of vitamins, in
treatment of rickets, Tuberculosis, and
also as a nutritive.
Storage
The oil should be stored in well-filled
airtight containers, protected from light,
and kept in a Cool place.
161

Wool Fat
Common Name: Lanolin, purified wool fat.
Biological Source: It is a yellow waxy substance
secreted by the sebaceous glands of wool bearing
animals like sheeps, Ovis aries. Family: Bovidae.
Geographical Location: Commercially it is
prepared in New Zealand, Australia, USA and
India.
162

Method of Preparation
Sheep’s wool contains about 45% of a fat
known as suint, which must be removed.
Crude lanolin is separated by washing
with sulphuric acid and then purified
and bleached.
The product is known as anhydrous
lanolin or wool fat.
Further the hydrous wool fat is
produced by intimately mixing wool fat
with 30% of water.
Physical properties
Colour : whitish yellow
Odour : slight and characteristic
Taste : bland
Solubility : insoluble in water but
forms turbidity with ether and
Chloroform
Melting-point : 40° to 44.4°c
Saponification value : 92 - 106
Acid value : less than 1
163

Wool fat contains the alcohols, cholesterol and
isocholesterol, together with various esters.
Hydrous wool fat also contains the acids in
combination with lanoceric, lanopalmitic,
carnaubic, myristic, oleic, cerotic and palmitic
acids.
It contains 50% of water.
164

Identification Tests
1. 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 color is produced,
indicating the presence
of cholesterol.
2. When a 2% solution in chloroform is
gently poured over the surface of
concentrated sulphuric acid, it
gradually develops a purple-red
colouration at the junction of the
liquids.
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.
165

Bees Wax
Synonyms: White beeswax, yellow beeswax, cera
alba, and cera flava.
Biological Source: Beeswax is the purified wax
obtained from honeycomb of hive bee, Apis
mellifera Linn and other species of Apis, belonging
to family Apidae.
Geographical Source: It is mainly found in Jamaica,
Egypt, Africa, India, France, Italy, California etc.
166

167
Preparation
The worker bee secretes the wax due to the ability of maintaining a high temperature
and the wax is secreted in the last four segments of abdomen on the ventral surface.
Just below the sterna it has a smooth layer of cells form the chitinous area that
secretes the wax.
The chitinous area has small pores through, which the wax exudes out.
The wax is passed to the front leg and later to the mouth; in the mouth it gets mixed
with the saliva, which is then built on the comb.
This wax forms a capping on the honey cells.
Wax forms about 1/8th part of the honeycomb. After removal of honey, honeycomb or
the capping is melted in boiling water.

168
On cooling the melted wax gets solidified and floats on the surface of water while
the impurities settle below and honey leftovers get dissolved in water.
The pure wax is then poured into earthen vessels wiped with damp cloth and the
wax so obtained is yellow beeswax.
White beeswax is obtained from yellow beeswax. The yellow beeswax is runned on
a thin stream of spinning wet drum, from which long ribbon like strips are scrapped
off.
The ribbon strips are placed on cloth in thin layers, rotated from time to time and
bleached in sunlight till the outer layer becomes white. White beeswax is obtained
by treating yellow beeswax chemically with potassium permanganate, chromic acid
or chlorine or charcoal.

Physical Properties
Beeswax consists primarily of a mixture of esters of fatty acids and fatty alcohols, paraffinic
hydrocarbons and free fatty acids.
Two types of beeswax are marketed — yellow beeswax and white beeswax.
Yellow beeswax is light-brown solid, brittle in nature when cold and presents characteristic
odour of honey.
White beeswax is a white or yellowish white solid having a characteristic, but faint, odour of
honey.
169

170
Yellow beeswax is smooth and soft to touch but breaks with granular fracture. It is insoluble in
water, but soluble in hot alcohol and other organic solvents.
In hot water it melts in liquid form and can be made any required shape and design after
cooling.
They are partially soluble in cold carbon disulfide and completely soluble in the same solution
at temperatures of 30° and above. Beeswax has a specific gravity of about 0.95.
Melting range : 60 - 65°C Acid value : 17 - 24
Peroxide value : Not more than 5 Saponification value : 87 - 104
Ester value : 72 - 79
The composition of beeswax depends to some extent on the subspecies of the bees, the age of
the wax and the climatic circumstances of its production, and hence the physical properties are
also varied.

171
Chemical Properties
Main constituents of beeswax are myricin (80%), cerotic acid, melissic acid and 15%
hydrocarbons.
Beeswax consists of five main groups of components:
1. Free fatty acids, most of them are saturated and have chain length of C24- C32.
2. Free primary fatty alcohols with a chain length of C28- C38.
3. Linear wax monoesters and hydroxymonoesters (35-45%) with chain lengths generally C40-C48.
The esters are derived almost exclusively from palmitic acid, 15-hydroxypalmitic acid and
oleic acid.
4. Complex wax esters (15-27%) containing 15-hydroxypalmitic acid or diols, which, through
their hydroxyl group, are linked to another fatty-acid molecule.
5. In addition to such diesters, tri and higher esters are also found.

Uses
Beeswax is used as a component in dietary food supplements (soft gelatin capsules and tablets),
glazings and coatings, chewing gum, water-based flavoured drinks, and as a carrier for food
additives (including flavours and colours) and cosmetics (Lipsticks, face creams).
Beeswax is suitable stabilizer for keeping oil-based capsule contents in suspension as well
as in tablet formulations.
Beeswax is blended with other oils and is used as a glazing agent for confectionery
(including chocolate), in small products of fine bakery ware coated with chocolate, in
snacks, nuts, coffee beans, dietary food supplements and in certain fresh fruits.
172

Adulterants:
Beeswax is adulterated with paraffin, microcrystalline wax, Jan wax, carnauba wax,
tallow and stearic acid.
They are identified by saponification values and as well as solubility and melting point
which is about 10-20°C lower than other waxes.
Beeswax will not give turbidity when boiled with sodium hydroxide and cooled, but
other waxes will form turbidity.
173

Marine flora and fauna play a significant role as a source of new molecular entity. The oceans
of the world contain over five million species in about 30 phyla.
Because of the diversities of marine organism and habitats, marine natural products enclose a
wide variety of chemical classes, including terpenes, shikimates, polyketides, acetogenins,
peptides, alkaloids of varying structures and a multitude of compounds of mixed biosynthesis.
During the last 30–40 years numerous novel compounds have been isolated from marine
organisms having biological activities such as antibacterial, antiviral, antitumour, antiparasitic,
anticoagulants, antimicrobial, antiinflammatory and cardiovascular active products.
175

The oceans cover more than 70% of the earth’s surface and contain over 200,000
invertebrates and algal species.
Although, numerous compounds have been isolated from marine organisms and the
biological activities attributed too many of them; but still very few of them have been
marketed or are under development. There are number of reasons that are why more number
of compounds originating from marine plants and animals has not been developed.
Macroalgae or seaweeds have been used as crude drugs in the treatment of iodine deficiency
states such as goitre, etc.
176

Some seaweed have also been utilized as sources of additional vitamins and in the
treatment of anaemia during pregnancy.
Marine products have also been used for the treatment of various intestinal disorders as
vermifuges, hypochloesterolaemic and hypoglycemic agent, e.g. Cystoseria barbata,
Sargassum confusam and Jania rubens.
Seaweeds have also been employed as dressing materials, ointments and in
gynaecology. For example, Porphyra atropurpurea have been used in Hawaii to dress
wounds and burns.
177

Durvillaea antractica to treat scabies in New Zealand.
Prepared, sterilized stripes of Laminaria digitala in conjunction with prostaglandins
have been used to dilate the cervix, as the strips swell up to several times to their
original diameter when moistened.
178

ANTIVIRALAGENTS
Name of drug Functional group source Action
Ara-A arabinosyl
nucleoside
Tethya crypta
marine sponge
semisynthetic antiviral
agent
Eudistomins β-caboline
derivatives
Eudistoma olivaceum,
sponges and gorgonians
family Polycitoridae
Also found in tunicates
Didemnins Trididemnum Spp. family
Didemnidae.
antiviral and
antitumour agents
182

183
Avarol and
Avarones
sesquiterpene
benzenoids
sponge
Disidea avara
anti-HIV activity
against the human
immunodeficiency virus
(HIV).
Patellazole B complex derivative ascidian
Lissoclinum patella
activity
against Herpes simplex
virus.
Fucoidan sulphated
polysaccharide
brown algae
Laminaria
activity against
HIV and Herpes simplex
viruses.

ANTICANCER AGENTS
cytosine
arabinoside,
(Ara-c)
spongothymidine Caribbean sponge
(Cryptotethya
crypta).
Treatment of acute myelogenous
leukaemia and human acute leukaemia.
potent inhibitor of the tumours in
cases of sarcoma-180, Erlich carcinoma
and L-1210 leukaemia in mice
Bryostatin I Bugula neritina
Bryozoal marine
organism
Crassin acetate cyclic diterpene
exocyclic lactone
Pseudoplexaura
porosa soft corals
184

Primary metabolites, Fibers, Teratogens, Marine organism

Primary metabolites, Fibers, Teratogens, Marine organism

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