Abstract

ANALYTICAL METHOD DEVELOPMENT, VALIDATION AND STRESS DEGRADATION STUDY OF DACLACTASVIR BY RP-HPLC METHOD

Amol V. Sawale*, Kalyani V. Tighare and Monika Jadhav

Abstract

To develop and validate simple, rapid, linear, accurate, precise and economical reverse phase-high-performance liquid chromatography (RP-HPLC) method for of stress degradation study of Daclatasvir. The separation and quantization were achieved on Inertsil C18(250 mm × 4.6ID,5 um). The mobile phase selected was Methanol: Water (80:20) at a flow rate of 0.8 ml/min and detection of analyte was carried out at 317nm at pH 3. The method exhibited good linearity over the range of 10–50 μg/mL. The drug is freely soluble in organic solvents Methanol. The drug was identified in terms of solubility studies and on the basis of melting point done by capillary tube method. The drug which when subjected to thermal, photolytic, oxidative, and acidic stress degraded into many degradation products. In most of the cases, the degradation rate was seen to be directly proportional to the amount of stress applied. The thermal stress was increased by increasing the incubation temperature, the faster the degradation took place. The values of LOD were found to be 0.845ug/ml for DCV. and the calculated LOQ values were found to be 0.054ug/ml. The low values of LOD and LOQ indicates the sensitivity of the method.

Keywords: Daclatasvir, Method development, Validation, Force degradation, RP-HPLC.