Method Development and Force Degradation Study for Daclatasvir Using LC-MS/MS

Abstract

Daclatasvir (DCV) used for the treatment of chronic hepatitis C virus infection (HCV). The present piece of work contains development, validation and force degradation study for DCV has been carried out by using LC-MS/MS technique. The LC-MS system contains 4000 QTrap along with the Shimadzu LC 20AD LC System, controlled by Analyst 1.4.2 software for present study. For the preparation of standard drug DCV and its marketed formulations the diluent used was acetonitrile: water (50:50v/v). LC system runs with the conditions like use of C18 column (Thermo Scientific, 5µm, 4.6 X 250 mm size), column temperature 25°C (Ambient), Injection volume 2 µL, total run time 10 min, flow rate 0.5 mL/min and an isocratic mobile phase consist of 1mM Ammonium acetate Buffer pH=4 adjusted with 50% acetic acid in water as ‘A’ and acetonitrile as a solvent ‘B’ (20:80 v/v). Validation was carried out by testing parameters like LOD (limit of detection), LOQ (Limit of quantification) linearity, ruggedness, accuracy and precision in terms of reproducibility, repeatability according to ICH guidelines. The calibration range for DCV was 500 to 5000 ng/mL, where 800, 1600 & 3200 ng/mL concentrations of DCV set as an Internal QC with proper labeling. All Three QC samples applied n=6 times. For Mass spectrometry (MS) quantification of the DCV, the interest of mass m/z 740.50/514.10 was considered. The force degradation study has been carried out on LC-MS for DCV at different stress conditions. The drug was found to be stable in acidic, neutral, oxidative and photolytic but degrades in basic conditions. The proposed method is fast, economical, avoiding chemical steps and wet chemistry and thus provides a comparatively simpler, rapid green method for routine analysis in pharmaceutical industries.