Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING UPLC METHOD FOR ESTIMATION OF APIXABAN AND ITS PROCESS RELATED IMPURITIES IN TABLET DOSAGE FORM

M. Madan Mohan Reddy*, D. Gowri Sankar and J. V. L. N. Seshagiri Rao

Abstract

A validated Ultra Performance Liquid Chromatography (UPLC) stability indicating method has been developed for determination of Apixaban (APB) and its process related impurities as per ICH guidelines in bulk and solid oral dosage forms in the presence of degradation products. Reversed-phase chromatography was performed on a UPLC BEH Waters C18 (50 x 2.1 mm; 1.7 m) with mobile phase consists of phosphate buffer (pH 3.0), Acetonitrile in a gradient proportion of 30:70 v/v respectively at a flow rate of 0.3 mL min-1. Detection was performed at 220 nm and a sharp peak was obtained for Imp-A, Imp-B and APB at a retention time of 0.862, 0.953, 1.325 minutes respectively. Linear regression analysis data for the calibration plot showed there was a good linear relationship between response and concentration in the range 10-60 μg/mL for APB with regression coefficient 0.9999 and for Imp-A and Imp-B is 0.625-3.75 μg/mL with the regression coefficients 0.9994 and 0.9999 respectively. The LOD values for APB and Imp-A & B were found to be 0.12 μg/mL, 0.06 μg/mL, 0.05 μg/mL & the LOQ values 0.40 μg/mL, 0.20 μg/mL, 0.16 μg/mL respectively. In order to determine whether the analytical method and assay were stability-indicating, APB and Impurities was stressed under various conditions to conduct forced degradation studies. Stability indicating forced degradation established studies showed results that there is no interference of any degraded products and there is no intererence from the excipients in the formulation. The detection of APB and the performed assay is thus specific stability- indicating. The wide linearity range, sensitivity, accuracy, short retention time, and simple mobile phase composition imply the method is suitable for routine quantification of APB with high precision and accuracy.

Keywords: Apixaban; Impurity A & B, Stability Indicating, UPLC.