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Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF SOFOSBUVIR AND DACLATASVIR DIHYDROCHLORIDE IN SOLID DOSAGE FORM

Khushbu S. Bhavsar*, Amit Khandhar and Dr. Paresh U. Patel

ABSTRACT

A rapid and sensitive stability indicating analytical method was developed for the estimation of Sofosbuvir and Daclatasvir dihydrochloride drug by RP-HPLC. The chromatographic separation was achieved on Zorbax Rx C8 (250*4.6 mm, 5 μm) at 35˚C. The separation is achieved employing a Mobile Phase – (A) consist of KH2PO4 Buffer (pH- 2.5): Acetonitrile (95:05) % v/v and Mobile Phase – (B) Acetonitrile: Methanol: Water (40:50:10) % v/v. Both mobile phases are used in the ratio of (50:50) % v/v. The flow rate was 1.0 ml/min. and detection was carried at 272 nm. The individual absorption maxima was found at 261 nm and 316 nm for Sofosbuvir and Daclatasvir dihydrochloride respectively. The average retention time was found to be 4 min. for Daclatasvir dihydrochloride and 12 min for Sofosbuvir. The proposed method was validated for system suitability, precision, linearity, accuracy, specificity, robustness. All validation parameter were within the acceptable limit. The assay method was found to be linear from 50% to 150% for Sofosbuvir and Daclatasvir dihydrochloride. Forced degradation studies on the formulation were conducted by adopting the proposed method to assess the stability of analyte under acid, alkaline, peroxide, humidity, thermal and photolytic condition and suitability of method to resolve degradation product.

Keywords: Sofosbuvir, Daclatasvir dihydrochloride, RP-HPLC Method, KH2PO4 Buffer (pH- 2.5).


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