Development and Validation of UPLC-MS/MS Method for the determination of Rivaroxaban in human plasma using Liquid-liquid extraction

A simple, sensitive, and selective ultra-performance liquid chromatographic (UPLC) method with mass spectrometric (UPLC-MS/MS) detection was developed for the determination of Rivaroxaban in human plasma sample. After a simple protein-precipitation by acetonitrile, the post-treatment samples were separated on a UPLC Bridged Ethyl Hybrid (BEH) C-18 column with 0.2% (v/v) formic acid in water: acetonitrile (60:40, v/v) as a mobile phase and analyzed in positive ion mode. The method developed was validated in human plasma with a daily working range of 2.02–997.72ng/mL with correlation coefficient, r2=0.999936. The method proved to be accurate (recovery, 97.0–107.95%), precise (2.21-7.21%), rapid (run time 2.5 min) and specific. In conclusion, the present validated method could be applied for pharmacokinetic study and bioequivalence testing of Rivaroxaban products.