A Stability-Indicating Ultra-Performance Liquid Chromatographic Method for Estimation of Related Substances and Degradants in Rivaroxaban Active Pharmaceutical Ingredient

Objective: The objective of the study was to develop and evaluate the reverse phase ultra-performance liquid chromatography (RP-UPLC) method for the quantitative determination of potential impurities of rivaroxaban active pharmaceutical ingredient. Method: The method uses a water acquity BEH C8 column (100mm x 2.1 mm, 1.7μm) with mobile phase A consisted, 0.05M diammonium hydrogen phosphate, pH adjusted to 3.0 and acetonitrile (80:20, v/v) and mobile phase B consisted mixture of acetonitrile and water (90:10, v/v) with a gradient programme. The column temperature was maintained at 30oC and the detection was carried out at 254nm. Results and Discussion: Efficient and reproducible chromatographic separation was achieved on BEH C8 stationary phase in gradient elution profile. The newly developed UPLC method was validated accordingly to ICH guidelines considering five impurities to demonstrate precision, linearity, accuracy and robustness of the method. The developed UPLC method was found to be rapid (15.0min runtime), accurate and sensitive. The correlation coefficient values are greater than 0.999 for rivaroxaban and its five impurities. Detection limit and Quantitation limit were 0.0005μg/mL and 0.0015 μg/mL respectively, indicating the high sensitively of the newly developed method. Accuracy of the method was established based on the recovery obtained between 98.4% and 103.5% for all impurities. The results of robustness study also indicates that the method is robust and is unaffected by small variation in chromatographic conditions. Conclusion: The proposed UPLC method provides reliable, reproducible, accurate and sensitive for the quantification of rivaroxaban related substances.