METHOD DEVELOPMENT AND VALIDATION OF ASSAY AND DISSOLUTION METHODS FOR THE ESTIMATION OF DACLATASVIR IN TABLET DOSAGE FORMS BY REVERSE PHASE HPLC.

V. Ashok Chakravarthy* and B.B.V. Sailaja

ABSTRACT

The objective of the present work is to develop a simple, efficient and reproducible stability indicating reverse phase high performance liquid chromatographic (RP-HPLC) method for the estimation of Daclatasvir (DCV) in tablet dosage forms for Assay and Dissolution methods. The developed method for Assay and Dissolution testing of Daclatasvir in tablets was carried out on Zorbax Eclipse Plus C-18 (100 x 4.6 mm, 3.5μm) column using Water and Methanol (20:80) by isocratic run. Flow rate was 1.5 mL min-1 with a column temperature of 25°C and detection wavelength was carried out at 315 nm. Total run time for the chromatographic analysis was 2 minutes for both assay and dissolution tests. The forced degradation studies were performed for assay content on Daclatasvir tablets under acidic, basic, oxidation, thermal, humidity and photolytic conditions. No degradation products were observed from the forced degradation studies. The method was validated in terms of specificity, linearity, accuracy, precision and robustness as per ICH guidelines. The method was found to be linear in the range of 10% to 150% of test concentration for assay content and dissolution rate respectively. The percentage recovery values were in the range of 98.6 to 99.8% and 98.0 to 102.6% for assay and dissolution rate at different concentration levels. Relative standard deviations for precision and intermediate precision results were found to be less than 2% and 5% for assay and dissolution rate. The results obtained from the validation experiments prove that the developed method is a stability-indicating method and suitable for routine analysis.

Keywords: Daclatasvir, Assay, Dissolution, Development, Validation, HPLC.