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Abstract

RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF GRAZOPREVIR AND ELBASVIR IN BULK AND ITS DOSAGE FORM WITH STRESS STUDIES

Rupa Lavyanya S.*, Bangaruthalli J., M. L. N. Renuka, P. Akhila and N. Arunkumar

ABSTRACT

Objective: The objective of this study was to develop and validate rapid, specific, sensitive, and precise reverse-phase high performance liquid chromatography (RP-HPLC) method for the quantitative determination of Grazoprevir and Elbasvir. No interference from any component of pharmaceutical dosage form was observed. Methods: The chromatographic separation was performed on Inertsil ODS column discovery (4.6 x 250mm, 5 μ particle size) the optimized mobile phase consists of 0.1% OPA: Methanol (50:50% v/v) with a flow rate of 1.0 ml/min and ultraviolet (UV) detection at 225 nm. Results: The retention times were 1.867 and 2.812 min for Grazoprevir and Elbasvir, respectively. Quantification is achieved with PDA detection at 254 nm over the concentration range of 50–150 μg/ml for Grazoprevir and for Elbasvir 25–125 μg/ml, with a correlation coefficient of 0.999 and 0.999, respectively. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines with respect to linearity, accuracy, precision, specificity, and robustness. Forced degradation study was extended out under acidic, alkaline, oxidative, photolytic, and thermal conditions to demonstrate the stability-indicating capability of the developed HPLC method. The degradation products were well resolved from the main peak, thus proved the stabilityindicating power of the method. The results of the analysis were validated statistically. Conclusion: The method is precise, accurate, linear, robust, and fast. The short retention time allows the analysis of a large number of samples in a short period of time and, therefore, should be cost-effective for routine analysis in the pharmaceutical industry.

Keywords: Grazoprevir, Elbasvir, High performance liquid chromatography, Development, Validation, Forced degradation.


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