DEVELOPMENT AND VALIDATION OF RP-HPLC-DAD STABILITY INDICATING ASSAY METHOD FOR THE DETERMINATION OF DESMOPRESSIN IN DESMOPRESSIN TABLETS

The aim of this research was to develop and validate a novel, rapid, selective, specific, accurate and efficient stability indicating RP-HPLC-DAD assay method for the quantification of Desmopressin in Desmopressin tablets. Various chromatographic parameters namely change in mobile phase, buffer and solvent composition, column stationary phase and oven temperature were studied. The elution of Desmopressin is achieved with improved peak shape under a set of gradient chromatographic condition: a reverse phase Hypersil BDS C18 (250mm x4.6mm, 5.0μm) column with a mobile phase A consisting of acetonitrile and mobile phase B is phosphate buffer pH 4.5 (25:75), flow rate 1.2 mL/minute, UV detection wavelength 220 nm. The chromatographic run time was 12 minutes with Desmopressin peak eluting at 6.7 minutes. The developed method was found to be specific and linear in the range of 5- 15 μg /mL (r2 = 0.9998). The good precision was achieved because the maximum RSD was 1.25%. Mean recoveries was 100.3%.The percent RSD was less than 2%. The method is robust with respect to changes in flow rate and wavelength. Solution stability evaluation indicated no evidence of degradation product. Standard solution was stable for 88 hours and sample solution was for 48 hours at room temperature. Forced degradation study of Desmopressin shown that peak was pure and there was no coeluting peaks when samples were assayed against reference. The developed method was validated and found precise, robust, accurate, linear, specific and stability indicating ensuring suitability of the method for quantification of Desmopressin in tablets. Method was successfully used for routine analysis of Desmopressin formulations