RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS DETERMINATION OF DARUNAVIR AND RITONAVIR IN COMBINED TABLET FORMULATION

Abstract

A simple, precise, accurate, and cost-effective reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of Darunavir and Ritonavir in combined tablet dosage form. The separation was carried out using a Thermo ODS C18 column (150 mm × 4.6 mm, 5 μm) with a mobile phase consisting of 0.01N KH₂PO₄ buffer (pH 3.1) and acetonitrile in the ratio of 40:60 v/v. The flow rate was 1.0 mL/min and detection was performed at 260 nm. The retention times of Darunavir and Ritonavir were found to be 2.6 and 3.4 minutes, respectively. The method exhibited good linearity in the concentration range of 30–180 μg/mL for Darunavir and 5–30 μg/mL for Ritonavir, with correlation coefficients of 0.999 for both. The percentage recoveries for Darunavir and Ritonavir were 99.06% and 98.68%, respectively. The method was validated as per ICH Q2 (R1) guidelines for accuracy, precision, specificity, linearity, robustness, LOD, and LOQ. The proposed method can be effectively applied for routine quality control analysis of Darunavir and Ritonavir in pharmaceutical formulations.