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1. SUBJECT : APA
KARNATAKA COLLEGE OF PHARMACY
TOPIC : BIOLOGICAL TEST & ASSAY 0F
ANTIVENOM
SUBMITTED BY: SUBMITTED TO:
ABHISHEK DR. C. SREEDHAR
M. PHARM 1ST SEMESTER PROF. AND HOD
DEPT. OF PHARMACEUTICAL ANALYSIS DEPT. OF PHARMACEUTICAL ANALYSIS
KARNATAKA COLLEGE OF PHARMACY KARNATAKA COLLEGE OF PHARMCY
BANGALURU BANGALURU
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2. Contents
 1. Venom
 2. Anti Venom
 3. Biological Test
 4. Assay of Anti Venom
 5. Uses of Anti Venom
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3. VENOM
 It is a poisonous substance secreted by animals such as snakes, spiders,
and scorpions and typically injected into prey or aggressors by biting or stinging.
 The snake venom is poisonous, typically the yellow color fluid stored in salivary
gland of venomous snake .
 The 10 to 15 mg of poisonous is deadly to humans adult.
 The toxin is delivered through a specially evolved venom apparatus, such as fangs or
a stinger, in a process called envenomation.
 There are over 3000 species of snakes, but only 15% of snakes are dangerous like
Krait, Pit viper, cobra & Resell viper etc.
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4. The venomous animal equipped with a set of
apparatus namely
1. venom gland.
2. fangs.
COMPOSITION;-
1. Proteins
2. Enzymes (i.e. Neurotoxins, Cardiotoxins )
• 3. Amino acids
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5. TYPES OF VENOMS
I. Neurotoxic
II. Cytotoxic
III. Hemotoxic
IV. Myotoxic
A. Neurotoxic
 It attacks on the nervous system.
 This mainly includes Alpha neurotoxins, Fasciculins & Dendrotoxins
 The fasciculins are attack to cholinergic neurons by destroying Ache so Ach
cannot be broken down & stay in the receptor.
e.g:- Venom of cobra , Kraits etc.…
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6. B. Cytotoxic
 Cytotoxic involve phospholipases, cardiotoxins.
 Phospholipases helps to split cell membrane and helps to digest and subdue prey.
 Cause localization symptoms, like blue black on the site of bite due to limited blood
circulation.
C. Hemotoxic
 The venom causes hemolysis or induce blood coagulation.
 It attacks cardiovascular, circulatory system & muscle tissues that directly leading to
heart failure.
D. Myotoxic
 The venom contains peptides that destroy muscle fiber proteins & result in
myonecrosis.
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7. Antivenom
 Antivenom, also known as antivenin, venom antiserum, and antivenom
immunoglobulin, is a specific treatment for envenomation.
 It is composed of antibodies and used to treat certain venomous bites
and stings.
 Antivenoms are recommended only if there is significant toxicity or a
high risk of toxicity.
 Antivenom are made by immunizing donor animals such as horse or
sheep with snake venoms .
 these animals have robust immune system & produce powerful
antibodies can be bind to snake venom component.
 The antivenoms are obtained by harvesting and than purifying the
antibodies from the plasma of donor animals.
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8. Types of Anti venoms
A. Monovalent
When they are effective against a single venom species.
B. Polyvalent
When they are effective against a range of species or several
different species at a time.
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9. PRODUCTION
OF
ANTIVENOM
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10. IDENTIFICATION TEST
 Produces almost immediate coagulation of blood and citrated human plasma.
 Mix the solution fraction from at least 0.6 mg with 1 ml of polyvalent antisnake
venom serum and incubate the mixture at 37°C for 30 min. Inject 0.5ml of mixture
intravenously into a group of mice weighing between 18 and 20 g.
 Observe the animals for 24hrs no animal dies.
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11. LETHAL DOSE(LD50)
 The toxicity of sanke venom is assessed by a toxicological test called lethal
dose .
 LD50 is used to determind the conc. Of toxin required to kill half the membrane
of a tested population of animal.
 Venom LD50 of 10 to 15 mg is deadly to human adult but within one bite snake
delivery may around 100-120 mg venom .
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12. Bioassay of Snake Venom
 Take 10 Swiss albino mice as test animal (weigh18-20g).
 Mice were maintained under standard temperature conditions (22°c) with a
regular 12hour light/12-hour dark cycle and allowed free access to standard
laboratory food and water.
 Diluted venom sample in saline is injected to mice 0.50 ml of each mice.
 Observe the animals for 24 hours.
 Not less than 3 and more than 8 mice die in 2-24 hrs.
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13.  If the number of deaths not in range, then change the dilution concentration of
venom in saline.
 The increase in antibody level was achieved by boosting treatment maintain a high
level of specific antibody against the venom antigen. ELISA has been used for
detecting venom antigen from different species of various animals.
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14. Uses of Antivenom
1. Medicines derived from neurotoxins are
used to treat brain injuries, strokes.
2. Hemotoxins are used to treat heart attack
& blood disorders.
3. Antivenoms are also used in Anticancer
activities.
4. Anticoagulant Properties.
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15. THANK YOU
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