STABILITY-INDICATING REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS ESTIMATION OF SELECTED DRUGS IN BULK AND FORMULATIONS WITH GREENNESS ASSESSMENT USING GREEN ANALYTICAL PROCEDURE INDEX

Abstract

Objectives: A comprehensive, verified, and dependable reversed-phase high-performance liquid chromatography method was created to assess velpatasvir and sofosbuvir in bulk and fixed-dose combo tablets.

Methods: The analysis was performed using a Waters Alliance 2695 high-performance liquid chromatography (HPLC) system equipped with an autosampler, a 2998 PDA detector, and Empower 2 software. Chromatographic separation was achieved on a BDS Hypersil C8 column (4.6 × 50 mm, 5 μm; Thermo Scientific). The mobile phase consisted of a 50:50 (v/v) mixture of potassium dihydrogen phosphate buffer (pH 3.5, adjusted with 0.1% orthophosphoric acid) and HPLC-grade acetonitrile. The detection wavelength was set at 288 nm, and the flow rate was maintained at 1.0 mL/min.

Results: In the concentration range of 2.5–15 μg/mL for velpatasvir and 20–120 μg/mL for sofosbuvir, the methodology consistently exhibits high correlation coefficients (R²) of 0.999, indicating exceptional linearity. Sofosbuvir exhibited a higher degree of precision than velpatasvir, with a recovery rate of 100.11% in contrast to 99.31%. To verify the precision and robustness (with a %RSD of <2%), the International Council for Harmonization Q2 (R1) principles were applied. Velpatasvir’s limits of detection/limit of quantification values were 0.01/0.04 μg/mL, whereas sofosbuvir’s were 0.13/0.40 μg/mL.

Conclusion: The observed degradation levels state the method’s specificity and applicability for quality control and stability testing, as they are within acceptable limits. The green analytical procedure index (GAPI) was employed to assess the method’s environmental impact. Applicability for quality control and stability testing was also assessed. The method’s environmental impact was evaluated using the GAPI.