Reversed-Phase Liquid Chromatographic Method for Determination of Daclatasvir Dihydrochloride and Study of Its Degradation Behavior

Abstract

A simple and selective reversed-phase stability-indicating liquid chromatographic method has been developed and validated for the determination of daclatasvir in drug substance and drug product. Daclatasvir was subjected to acidic, alkaline, oxidative, thermal and photo-degradation study. The LC method was based on isocratic elution of daclatasvir and its degradation products on a reversed-phase C18 Hypersil column using a mobile phase consisting of phosphate buffer (10 mM, 1 mL triethylamine L−1): acetonitrile (60:40 v/v) at a flow rate of 2 mL min−1. Quantitation was achieved with UV detection at 312 nm. Linearity, accuracy, and precision were found to be acceptable over the concentration range of 0.75–120 μg mL−1, with regression coefficient value of 0.9999, and with limit of detection and quantitation of 0.148 and 0.447 μg mL−1, respectively. Peak purity was checked for principle drug and its alkali induced degradation product, and the pathway of alkaline hydrolysis of daclatasvir was suggested by LC/MS.