Abstract

DEVELOPMENT AND VALIDATION OF UV SPECTROPHOTOMETRIC METHOD AND HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHIC METHOD FOR ESTIMATION OF SOFOSBUVIR IN PHARMACEUTICAL FORMULATIONS

Ashwini A. Wakchaure* and Shubha P. Katte

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Abstract

Simple, rapid, precise, economical and specific UV spectrophotometric and high- performance thin-layer chromatographic (HPTLC) methods have been developed for quantitative analysis of Sofosbuvir (SFR) in pharmaceutical formulation. For UV spectrophotometric method, a mixture of methanol and water was used as a solvent. Initial stock solution of SFR was prepared in methanol and subsequent dilutions were done in water. The standard solution of SFR in water showed absorption maxima at 261nm. The drug obeyed Beer–Lambert’s law in the concentration range of 10–50 μg/mL with coefficient of correlation (R2) of 0.999. The recovery was obtained with values close to the 99.41% at three different concentrations, which reflect that the method is free from interference of the impurities and other additives used in tablet formulation. For HPTLC method, aluminum plates precoated with silica gel G60F254 was used as the stationary phase. The solvent system consisted of toluene: chloroform: ethanol in the proportion of 4:4:1, v/v/v. This solvent system was found to give compact spots for Sofosbuvir with RFvalue 0.30 ± 0.01. Densitometric analysis of Sofosbuvir was carried out in the absorbance mode at 261 nm. Linear regression analysis showed good linearity (r2 =0.998) with respect to peak area in the concentration range of 100–800 ng/spot. The both methods were validated for precision, limit of detection (LOD), limit of quantitation (LOQ), accuracy and specificity. Statistical analysis proved that the methods are repeatable and specific for the estimation of the said drug.

Keywords: Sofosbuvir, Spectrophotometry, HPTLC, Method Validation.