ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF ELBASVIR AND GRAZOPREVIR IN BULK AND TABLET FORMULATIONS BY RP- HPLC

Abstract

The aim of the study is to develop and validate a simple, accurate, precise and rapid isocratic reverse-phase highperformance liquid chromatographic method for simultaneous determination of Elbasvir and Grazoprevir in bulk and tablet formulations. Chromatographic separation was supported on Inertsil ODS, 5µm C18(150x4.6 ID) column with a blend of methanol: water (80:20) as mobile phase at a flow rate of 1 ml/min. UV detection was performed at 260 nm using HPLC Shimadzu (LC 20 AT VP) with LC solutions software. The retention times were found to be 7.15 minutes and 5.30 minutes for Elbasvir and Grazoprevir respectively. Calibration plots were linear for both the drugs (r2=0.999) over the concentration range 36-84 μg/ml for Elbasvir 6-14 μg/ml for Grazoprevir. The LOD and LOQ values for Elbasvir and Grazoprevir were found to be 0.81 µg/ml, 2.46µg/ml and 0.53µg/ml, 1.633µg/ml respectively. The optimized method was validated in accordance with ICH guidelines for accuracy, precision, specificity, linearity, and sensitivity. The proposed method was successfully used for quantitative analysis of Zepatier tablets. No interference from any component of pharmaceutical dosage form was observed. Validation studies revealed that method is specific, rapid, reliable, and reproducible. As the method shows high recovery and low relative standard deviation which confirms the suitability of the method for routine determination of Elbasvir and Grazoprevir in bulk drug and tablet dosage forms.