Study on fluorescence properties of HCV antiviral (velpatasvir) and its fluorimetric determination in presence of sofosbuvir; application to stability study and human plasma.

Velpatasvir (VLP) is a new, oral, direct-acting antiviral with potent inhibitory activity against all hepatitis C virus (HCV) genotypes. A highly sensitive, simple, fast and specific one fluorometric method for determination of VLP in the presence of sofosbuvir was developed and validated. The fluorescence behavior of VLP in different organic solvents was examined and explained. Methanol was concluded to be the best sensitizing reagent. The native fluorescence intensity of VLP was accomplished at 383 nm with 339 nm for excitation wavelength. The impacts of experimental variables included pH, various organized media, and time of stability were examined and optimized. A linear relationship was achieved between the VLP concentration and the fluorescence intensity in a range of 5 to 5 × 10 ng mL with 0.70 and 0.23 ng mL , for quantitation and detection limits respectively. The proposed method was utilized for analyzing of VLP in human plasma and additionally expanded to examine the stability of VLP after its exposure to various stress conditions, like oxidative, alkaline, acidic, UV, daylight and sunlight conditions, according to ICH guidelines. Furthermore, the kinetics of acidic and oxidative degradations of VLP was examined. Moreover, the half-life times of the reaction (t ) and the first-order reaction rate constants were estimated. Finally, a suggestion for the degradation pathway was presented.