ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF ABALOPARATIDE AND TERIPARATIDE IN COMBINED DOSAGE FORM BY USING RP-HPLC METHOD

New method was established for simultaneous estimation of Abaloparatide and Teriparatide by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Abaloparatide and Teriparatide by using ACE C18 column (4.6x150mm) 5 mu, flow rate was 1.2 ml/min, mobile phase ratio was (70:30 v/v) methanol: Phosphate buffer pH 3 (pH was adjusted with orthophosphoricacid), detection wavelength was 240nm. The instrument used was Shimadzu, model No. SPD-20MA LC+20AD, Software-LC-20 Solution. The retention times were found to be 2.344 mins and 3.284 mins. The % purity of Abaloparatide and Teriparatide was found to be 101.27% and 99.97% respectively. The system suitability parameters for Abaloparatide and Teriparatide such as theoretical plates and tailing factor were found to be 4668, 1.3 and 6089 and 1.2, the resolution was found to be 6.0. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study n Abaloparatide and Teriparatide was found in concentration range of 50 mu g-250 mu g and 5 mu g-50 mu g and correlation coefficient (r(2)) was found to be 0.999 and 0.999, % recovery was found to be 99.56% and 99.48%, %RSD for repeatability was 0.2 and 0.2, % RSD for intermediate precision was 0.2 and 0.1respectively. The precision study was precise, robust, and repeatable. LOD value was 3.17 and 5.68, and LOQ value was 0.0172 and 0.2125 respectively. Hence the suggested RP-HPLC method can be used for routine analysis of Abaloparatide and Teriparatide in API and Pharmaceutical dosage form.