Bio assay of oxytocin

Bio assay of oxytocin
Balaji college of pharmacy
Mr.B.CHAKRAPANI M.pharm
ASSISTANT PROFESSOR
PHARMACOLOGY&clinicalpharmacolgy
Balaji college of pharmacy
Phone no :+91-9618279507
anticancer@rediffmail.com
anantapur.

Principle involved in oxytocin
The potency of oxytocin is determined by
comparing its activity with that of the
Standard Preparation of oxytocin under the
conditions of a suitable method of assay.
8/3/2017 2STRENGTH IS LIFE WEAKNESS IS DEATH....

8/3/2017 strength is life weakness is death.... 4

Standard Preparation
The Standard Preparation is the 4th
International Standard for Oxytocin,
established in 1978.
It consisting of freeze-dried synthetic
oxytocin peptide with human albumin and
citric acid (supplied in ampoules containing
12.5 Units), or another suitable preparation
the potency of which has been determined
in relation to the International Standard.
8/3/2017 9strength is life weakness is death....

Suggested methods
Method A:
By depression of the blood pressure in
chicken
Method B:
By contraction of the rat uterus
Method C:
By measurement of milk-ejection pressure in a
lactating rat .

Method B
By contraction of the rat uterus
Inject 100 mcg of oestradiol benzoate .
Intramuscularly into a female rat weighing 120
to 200 g 18 to 24 hours. before the assay.
Immediately before the assay confirm by
vaginal smear that the rat is in oestrus or
preoestrus.
Kill the rat and suspend one horn of the uterus
in a bath containing a solution of the following
composition:

Sexual dimorphismBALAJICOLLEGEOFPHARMACYATP

Kill the rat and suspend one horn of the uterus in a bath
containing a solution of the following composition:
Composition (% w/v)
Sodium chloride 0.662
Potassium chloride 0.045
Calcium chloride 0.007
Sodium bicarbonate 0.256
Disodium hydrogen phosphate 0.029
Sodium dihydrogen phosphate 0.003
Magnesium chloride 0.010
Dextrose 0.050

Maintain the bath at a temperature of 32o or at some
other suitable temperature at which spontaneous
contractions of the uterus are abolished and the
preparation maintains its sensitivity.

Oxygenate the solution with a mixture of 95%
of oxygen and 5% of carbon dioxide and
record the contractions of the muscle using a
suitable instrument giving a linear response
(for example an isotonic lever with a load not
exceeding 2 g).

Isotonic lever

Record the contractions produced by the
addition to the bath of two doses of the
Standard Preparation suitably diluted with the
above solution.
The doses should be such as to produce clearly
discriminated, sub maximal contractions; the
required doses normally lie between 10 and 50
micro Units per ml of bath liquid.

When maximal contraction has been reached,
replace the bath liquid by a fresh solution.
The doses should be added at regular intervals
of 3 to 5 minutes depending upon the rate of
recovery of the muscle.
Dilute the preparation being examined so as to
obtain responses on the addition of two doses
similar to those obtained with the Standard
Preparation.

The ratio between the two doses of the
preparation being examined should be the
same as that between the two doses of the
Standard Preparation and this ratio should be
kept constant throughout the assay.

The two doses of Standard Preparation and the
two doses of the preparation being examined
should be given according to a randomized
block or a Latin square design and at least six
responses to each should be recorded.
Measure all the responses and calculate the
result of the assay by standard statistical
methods.

THANK YOU

Bio assay of oxytocin

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Bio assay of oxytocin