Development and Validation of a Stability-Indicating Reversed Phase High-Performance Liquid Chromatography Method for Quantifying Rivaroxaban (XARELTO): Synergistic Application of Infrared Spectroscopy and Clinical Pharmacology Insights

Aktham Mestareehi ^1,2,3*

• ¹ Wayne State University, Detroit, United States
• ² Northeastern Illinois University, Chicago, Illinois, United States
• ³ Isra University, Amman, Amman, Jordan

Rivaroxaban is an anticoagulant that targets Factor Xa, a key enzyme in the blood clotting cascade, to prevent clot formation and growth. It is widely used to reduce the risk of stroke and systemic embolism in patients with non-valvular atrial fibrillation, treat deep vein thrombosis (DVT) and pulmonary embolism (PE), and prevent recurrent DVT and PE. Additionally, it serves as prophylaxis against DVT in patients undergoing hip or knee replacement surgery.A robust, precise, and selective reversed-phase high-performance liquid chromatography (HPLC) method was developed and validated for Rivaroxaban analysis in raw materials. The method utilized isocratic elution at a flow rate of 1 mL/min on a Thermo ODS Hypersil C18 column (4.6 × 250 mm, 5 µm) at ambient temperature. The mobile phase consisted of monobasic potassium phosphate buffer (pH 2.9) and acetonitrile (70:30, v/v), with UV detection at 249 nm.The method demonstrated excellent linearity over a concentration range of 50–1000 ppm (R² = 0.999), with a retention time of approximately 12 minutes. Precision and accuracy, indicated by percentage relative standard deviation (RSD), remained consistently below 2.0%, confirming method reliability. Solution stability studies confirmed Rivaroxaban’s stability throughout the analysis, with no peak loss, degradation, or additional peaks observed between the first and last injections.Forced degradation studies were performed under various stress conditions, including acid and base hydrolysis and hydrogen peroxide oxidation. The method successfully separated Rivaroxaban from its degradation products, confirming its stability-indicating capability.Validation followed International Conference on Harmonisation (ICH) and U.S. Pharmacopeia (USP) guidelines, assessing specificity, robustness, linearity, accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ). The LOD for impurities and degradants was determined at 0.3 ppm, while the LOQ was 1 ppm.This validated, stability-indicating method is highly suitable for routine quality control and analytical applications, offering simplicity, efficiency, and robustness in both raw materials and finished pharmaceutical products.