Development and Validation of Analytical Method for Assay Determination of Isosulfan Blue by Liquid Chromatography
A novel, simple and rapid method has been developed and validated for quantitative assay determination of
isosulfan blue in drug substance by Ultra Performance Liquid Chromatography (UPLC). Analytical method was developed
and validated as per ICH guidelines. Reverse phase liquid chromatography method is developed and validate on UPLC using
C18 (100 x 2.00 mm) 1.9μ column. 0.1 % perchloric acid in mixture of 73 volumes of water and 27 volumes of acetonitrile
as mobile phase. The ultra violet wavelength selected 220 nm, the column temperature 30ºC. Linearity study was carried out
for isosulfan blue linearity was calculated from 80% level to 120% with respect to specification level. The correlation
coefficient r = 0.999 was proved and showed that the method is linear. Specificity was studied by individual injection of
drug substance, related impurities and spiking study as well. It was evident from specificity test that all the related impurities
are separated from isosulfan blue peak and method is specific. Isosulfan blue sample solution and mobile phase were found
to be stable for at least 3 days.
Keywords - Isosulfan blue, UPLC, Assay, Validation, Reverse phase, Solution stability.