Stability-indicating HPLC method development and validation of rivaroxaban impurities and identification of forced degradation products using LC-MS/MS.

Abstract

The current aim of this study is to develop a simple, sensitive, and robust analytical method that can separate the Rivaroxaban and its related impurities by using HPLC. This new method can separate enantiomers and all the process-related impurities of rivaroxaban. This method can also be used to determine the assay of rivaroxaban in drug substances and drug products. This new method was developed using a Chiralpak IC (250 × 4.6mm, 5μm) column at 27°C with a gradient program of 25.0min at a flow rate of 0.8mL/min. Mobile phase A consisted of acetonitrile, ethanol, and n-butyl amine in the ratio of 95:5:0.5 (v/v/v), respectively. Mobile phase B consisted of a mixture of Milli-Q water, methanol, and n-butyl amine in the ratio of 50:50:0.5 (v/v/v), respectively. The detector wavelength was 254 nm. Rivaroxaban was subjected to the forced degradation conditions (stress conditions) of hydrolysis, base, acid, thermal, photolysis, and oxidation. The degradation products were well separated from rivaroxaban peak and enantiomer peak, and its potential impurities prove the stability-indicating power of the method. The proposed new stability-indicating method was validated with respect to specificity, precision, accuracy, limit of quantification, limit of detection, linearity robustness, and roundness per International Conference on Harmonization guidelines. The %RSD (relative standard deviation) for six sample preparations for rivaroxaban and impurities less than 10% and the recovery is between 90 and 110%. The current method can be used in quality control labs for analysis of commercial products.