Amprolium Hydrochloride (Finished product) Assay (By HPLC)
Mobile Phase - Buffer : Methanol : ACN ( 500 : 450 : 50 )
Chromatographic condition:
Column
- C18 250 x 4.6 mm ( 5 um ) Make : Nucleodur
Flow Rate - 1.0 ml / minute.
Diluent - Mixture of 500 ml of water, 450 ml of methanol, and 50 ml of acetonitrile.
Wavelength - 254 nm
Injection Volume - 20 µl
Run Time - 25 Minute
Retention Time - About 14 Minute
Buffer Preparation - Dissolve 6.0 g of sodium 1-heptanesulfonate in 500 ml of water, add 12 ml of glacial acetic acid, 2 ml of triethylamine, 450ml of methanol, 50ml of acetonitrile and mix.
Standard Preparation: Take 50 mg of Amprolium HCl WS in 100 ml volumetric flask and add diluent to dissolve, and make up volume upto mark with diluent.
Sample Preparation: Weigh powder equivalent to about 50 mg of Amprolium HCl, in 100 ml volumetric flask, add 30ml of diluent and sonicate for 15 min and make up volume upto mark with diluent. Shake and filter.
Procedure : Inject one blank , five replicate injection of standard solution, two
replicate injection of sample solution & two bracketing
standard solution.
System Suitability :
1) RSD of
five replicate injection of standard solution should not be more than 2.0%
2) RSD of five replicate injection of standard solution and two bracketing standard solution should not be more than 5.0%
3) Tailing factor should not be more than 2.0
4) Theoretical plates should not be less than
2000
Calculation :
