Ivermectin Injection Assay (MOA) (By UV HPLC)
Mobile Phase : Water : Acetonitrile :
Methanol (7:58:35 v/v)
Chromatographic conditions :
Column - C18(250 mm x
4.6 mm) 5 µ or equivalent.
Detector
and Wavelength - VWD and 254 nm
Flow rate - 1.0 ml/min
Injection
volume - 10 µl
Column
temperature - 30°C + 2°C
Auto
sampler temperature - 15°C + 1°C
Run Time - 25 min
Retention
Time - Ivermectin B1a - about 5.1 min and Ivermectin B1b - about 7.4 min
Diluent
and wash vial - Methanol
Standard Preparation
Accurately
weigh and transfer about 30.0 mg of Ivermectin
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.
Sample Preparation
Weigh and
transfer sample solution i.e. equivalent to 30 mg of Ivermectin in 100 ml volumetric flask, add about 70 of diluent,
sonicate for 6 min with intermittent shaking in every after 2 min and make up
the volume 100 ml with diluent.
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 2.0%
3) Tailing
factor should not be more than 2.0
Calculation
