Stress degradation studies and development of validated stability indicating assay method by RP-HPLC for estimation of Apixaban in presence of degradation products as per ICH guidelines

Abstract

A simple, selective, reproducible, economic, stability indicating method has been developed and validated as per ICH guidelines for the estimation of Apixaban (APB) both in bulk drug and in solid dosage forms in the presence of degradation products. Reversed-phase chromatography was performed on a Inertsil ODS (4.6 x 150mm, 5 mm) with mobile phase Octo sulphonic acid adjusted with Orthophosphoric acid to pH 3.0 with 300 ml (30%) and 700 ml of Acetonitrile HPLC (70%).at a flow rate of 1.0 ml min-1. Detection was performed at 226 nm and a sharp peak was obtained for APB at a retention time of 2.235 minutes. Linear regression analysis data for the calibration plot showed there was a good linear relationship between response and concentration in the range 10-50 ppm; the regression coefficient was 0.9997 and the Limit of detection (LOD) was 0.33 μg/mL with S/N ratio of 2.99 and limits of quantification (LOQ) was 30 μg/mL with S/N ratio of 9.97 respectively. In order to determine whether the analytical method and assay were stability-indicating, APB 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 it did not  interfere with excepients 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 imply the method is suitable for routine quantification of APB with high precision and accuracy.