ANALYTİCAL METHOD DEVELOPMENT AND VALİDATİON OF STABİLİTY INDİCATİNG RP-HPLC METHOD FOR THE SİMULTANEOUS ESTİMATİON OF SOFOSBUVİR AND DACLATASVİR İN BULK AND PHARMACEUTİCAL DOSAGE FORM

Abstract

Special, effective high pressure liquid chromatography method has been developed for the simultaneous quantification
of Sofosbuvir and Daclatasvir. By using Waters HPLC e-2695 quaternary pump with a PDA detector of 2998
instrument the chromatographic separation of Sofosbuvir and Daclatasvir was achieved on the column of Waters XBridge C18 150X4.6mm, 3.5µ using an isocratic elution with a buffer containing 0.1percent ortho phosphoric acid
and acetonitrile at a rate of 70:30 as a mobile phase with a flow rate of 1 ml/min at ambient temperature. A detector
wavelength of 265 nm utilizing the PDA detector were given in the instrumental settings. The linearity was studied
between the concentration range of 40-600 µg/ml of Sofosbuvir and 6-90 µg/ml of Daclatasvir were injected. The
plotted calibration curves were linear with a regression coefficient of R2> 0.999, indicates that the linearity was with
in the limit. As a part of method validation the parameters like specificity, linearity, accuracy, ruggedness, robustness
were determined and the results were found to be within the allowable limit. The method developed was found to be
applicable to routine analysis and to be used for the measurement of both active pharmaceutical ingredients (i.e,
Sofosbuvir and Daclatasvir). Validation of the proposed method was carried out according to an International
Conference on Harmonization (ICH) guidelines. Since, there is HPLC method reported in the literature for the
estimation of Sofosbuvir and Daclatasvir, there is a need to develop quantitative methods under different conditions
to achieve improvement in specificity, selecivity etc.