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Application of green solvents for developing green analytical methodologies has grown dramatically in the past few years. The “more hazardous reagents” are replaced by more “environment-friendly solvents” without affecting method performance. In the present study, two simple and accurate chromatographic methods were developed and validated for determination of the new antiviral combination sofosbuvir (SBR) and ledipasvir (LPV). The first adopted method is high-performance thin-layer chromatography coupled to densitometric determination where silica gel 60 F254 plates were used as the stationary phase. Whereas, the running mobile phase used was toluene: ethanol: ammonia (4:1:0.2, v/v/v). Reversed-phase high-performance liquid chromatography with ultraviolet detection was the second method developed. The column used was Inertsil C18 column (150 × 4.6 mm, 5 μm) and the mobile phase was 20 mM potassium dihydrogen orthophosphate (adjusted to pH = 3 using acetic acid): ethanol (60:40, v/v) with a flow rate of 1.0 mL/min. The detection wavelength for both methods was 265 nm. The validation of both methods was done according to ICH guidelines where both methods were found to be accurate, reproducible, and selective. The linearity range for HPTLC and RP-HPLC methods were 0.8–25.6 and 0.4–12.8 μg/band and 6.0–100.0 and 4.0–80.0 μg/mL for sofosbuvir and ledipasvir, respectively. Comparison of the developed methods was done with reported HPLC method where no significant difference was found.

Simultaneous chromatographic analysis of Sofosbuvir/Ledipasvir in their combined dosage form: an application to green analytical chemistry