Novel determination of a new antiviral combination; sofosbuvir and velpatasvir by high performance thin layer chromatographic method; application to real human samples

摘要

Sofosbuvir (SOF) and velpatasvir (VEL) represent a brand new antiviral combination. A novel, simple and economical high performance thin layer chromatographic separation coupled with densitometric quantification was designed for the pharmaceutical and biomedical analysis of SOF and VEL. Simultaneous quantitation of the studied drugs in their newly introduced pharmaceutical formulation and human plasma; using ledipasvir (LED) as an internal standard; was achieved. Protein precipitation technique using acetonitrile was adopted, utilizing HPTLC as a clean-up step. Separation was achieved on HPTLC silica gel 60 F-254 aluminum plates with a green developing system consisting of ethyl acetate-isopropanol (90:10, v/v). Densitometric scanning was carried out at 260 and 302 nm for SOF and VEL, respectively. Peak purity was evaluated through the winCATS (R) software spectral correlation method to ascertain the specificity of the proposed method. Method validation was assessed as per the US-FDA guidelines. The proposed method represents a simple, green and cost-effective alternative to the only existing more complicated reported LC-MS/MS technique. The method would afford a competent tool for therapeutic drug monitoring and bioavailability studies of SOF and VEL. The method could be applied to the quality control and routine analysis of SOF and VEL in their pure forms and pharmaceutical formulations. Furthermore, the application was extended to simultaneous quantitation of SOF and VEL in real human plasma.