Abstract
A simple, specific, accurate and stability-indicating reversed phase high performance liquid Chromatographic method was developed and validated for determination of related substances in Tazarotene gel. Chromatographic separation was Waters symmetry 150 × 3.9mm, 5μ column as stationary phase while mobile phase A was buffer: organic modifier, 40:60 v/v (Buffer=10 mM KH2PO4 with pH 3.0 by orthophospheric acid and organic modifier = methanol: tetra hydrofuran, 95:5 v/v) and mobile phase B as organic modifier. Method was developed in gradient mode with 55 minutes, at flow rate of 1.0 mL/min. Effluents were monitored at 325 nm. The method was validated for specificity, linearity, accuracy, precision, limit of quantification, limit of detection, robustness and solution stability. The RRF (relative response factor) values of impurity-A, impurity-B, impurity-C determined from linearity study were 0.99, 1.78 and 1.24 respectively. Limit of quantification of Tazarotene, impurity-A, impurity-B and impurity-C was found to be 0.02 µg/mL, 0.02 µg/mL, 0.01 µg/mL and 0.01 µg/mL respectively. Recovery was found to be in the range of 95-105%. The method was proved to be robust with respect to changes in flow rate, buffer pH and column temperature. The proposed method was successfully applied for the quantitative determination of related substances in Tazarotene topical formulation.
