In Vitro free Radical Scavenging Activity of Various Extracts of Aerial Parts of Dyschoriste littoralis Nees

摘要

A new, rapid, simple, precise and accurate stability-indicating reversed phase-HPLC method has been developed and validated for quantitative determination of daclatasvir (DAC) in pure form and tablets. An isocratic HPLC method, using Thermohypersile BDS C18 reversed phase column (150 mm × 4.6 mm i.d., particle size 5 μm) with isocratic ternary mobile phase consisting of methanol: acetonitrile: phosphate buffer (pH 3.0) (30:30:40, V:V:V), was investigated to separate DAC from its stress degradation products. The flow rate was 1.5 mL min-1 at ambient temperature and UV detector was used at 305 nm for detection. The elution time of DAC was found to be 2.195± 0.045 minutes. The developed method was validated for system suitability, linearity, accuracy, precision, limits of detection and quantitation, specificity, stability, robustness and for system suitability parameters as per ICH guidelines. The calibration curve was found to be linear with the equation y=51.399x + 4.8876, with a correlation coefficient of (R2=0.9999) over a concentration range of 2.0–120 μg mL-1. The limits of detection and quantification were 0.12 and 0.5 μg mL-1, respectively. The recovery value of this method is 99.0% and the reproducibility is within 0.90%. Stability tests were done through exposure of the analyte solution for five different stress conditions: Reflux with 1.0 mol L-1 hydrochloric acid (HCl), reflux with 1.0 mol L-1 sodium hydroxide (NaOH), reflux with 30% hydrogenperoxide (H2O2), exposure to ultraviolet radiation (UV) radiation and thermal conditions. The proposed method could be used for routine analysis of DAC in tablets.