BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF RIVAROXABAN IN HUMAN PLASMA BY RP-HPLC

Pratiksha Sunil Deshmukh*, Sugriv R. Ghodake and Hemant Vinayak Kamble

ABSTRACT

A simple, precise, and accurate reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the quantitative estimation of Rivaroxaban in human plasma. The method employed protein precipitation for sample preparation and used a mobile phase composed of acetonitrile and ammonium formate buffer (78:22 v/v) at pH 4.5. Chromatographic separation was achieved on a Symmetry C18 column (250 mm × 4.6 mm, 3.5 μm) with a flow rate of 1 mL/min, and detection was carried out at 251 nm. The retention time of Rivaroxaban was approximately 4.4 minutes. The method demonstrated good linearity in the concentration range of 5–25 μg/mL with correlation coefficients (r²) exceeding 0.995. Validation parameters including accuracy, precision, recovery, and stability were within the limits established by USFDA guidelines. Stability studies confirmed that Rivaroxaban was stable under various conditions, including freeze-thaw, short-term, long-term, and stock solution storage. The developed method is suitable for pharmacokinetic studies and therapeutic drug monitoring of Rivaroxaban in human plasma.

Keywords: Rivaroxaban, RP-HPLC, Bioanalytical Method, Method Validation, Human Plasma, Protein Precipitation, Acetonitrile, Ammonium Formate Buffer, Linearity, Accuracy, Precision, Recovery, Stability Studies, USFDA Guidelines, Pharmacokinetics etc.