Development of Validated Bioanalytical RP-HPLC Method for Determination of Sofosbuvir In Human Plasma

A linear, precise, accurate and simple reversed-phase high performance liquid chromatography (RP-HPLC) bioanalytical method was developed and validated for determination of sofosbuvir in human plasma (K2EDTA) using ledipasvir as an internal standard. Sofosbuvir and ledipasvir (ISTD) were separated from plasma using solid phase extraction technique and The chromatographic separation was accomplished by using a Zorbax eclipse XDB C18 Column (2504.6 mm i.d, 5 m particle size) with 1% orthophosphoric acid (pH6.4) and Acetonitrile in the ratio of 30:70% v/v, as mobile phase in an isocratic elution mode pumped at a flow rate 0.7 mL/min and the column oven temperature was maintained at 25C. The wavelength selected for quantitation was 254 nm.