Closantel sodium Injection Assay (MOA) By HPLC
Chromatographic Conditions :
Column - C18( 50 x
4.6mm ) 5μm or equivalent
Detector
wavelength - 240 nm
Flow Rate - 1.0 ml / Minute
Injection volume - 10 μl.
Retention time - About 16.0 min
Diluent
and wash vial - Methanol
Buffer Preparation:
Dissolve 7.7 gm ammonium
acetate in 1000 ml of water adjusted to pH 4.3 with acetic acid.
Filter with 0.45µm nylon membrane filter.
Gradient Progamme:
|
Time |
Mobile
Phase A |
Mobile
Phase B |
|
0 |
50 |
50 |
|
2 |
50 |
50 |
|
22 |
20 |
80 |
|
27 |
20 |
80 |
|
30 |
50 |
50 |
|
35 |
50 |
50 |
Standard Preparation
Accurately weigh and transfer about 52.0 mg of Closantel sodium Dihydrate WS in 100
ml volumetric flask and add about 70 ml of diluent, Sonicate to dissolve and
make up the volume 100 ml with diluent.
Dilute 5 ml of this solution to 50 ml with diluent
and mix.
Sample Preparation
Weigh and transfer sample solution i.e.
equivalent to 100 mg of Closantel in
100 ml volumetric flask, add about 70 of diluent, sonicate for 10 min with
intermittent shaking in every after 2 min and make up the volume 100 ml with
diluent.
Dilute 5 ml of this solution to 100 ml with diluent
and mix. Filter with 0.45µ nylon syringe 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 and Theoretical plates should not be less
than 2000.
Calculation :
