ASSAY (By HPLC)

Chromatographic Conditions:
Column : 12.0-cm × 4.6-mm column that contains 5-µm packing L1,
Wave Length : 220 nm
Flow Rate : 1.0 ml/minutes
Injection Volume : 20ul.
Mobile phase A:
Prepare a buffer solution of 0.1 M monobasic sodium phosphate.

Mobile phase B:

Prepare a filtered and degassed mixture of acetonitrile in water (1:1).

Diluents:

Dissolve 5.0 g of chlorobutanol in 5.0 mL of glacial acetic acid; add 5.0 g of

alcohol, 1.1 g of sodium acetate, and 1000 mL of water, and mix.

Standard preparation:

Weigh 8.350mg of working standard of OXYTOCIN in 100ml Amber color

volumetric flask, dissolve in mobile phase (Protect from light) transfer 2ml in to
100ml volumetric flask and make up with mobile phase.

Sample preparation:

Take 10 ml of sample solution in to 50ml Amber color volumetric flask and

make with mobile phase.

(Protect from light)

Procedure:

Inject 20µL volume of std. and test solutions in to a chromatograph and record
the chromatogram.

CALCULATION:

Mcg/ml = Area of sample x Std. Wt x 50 x P x 1 x1000

Area of Standard x 100 x Spl Vol. x 100

%age = Mcg/ ml x 100

L.C