THE ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF METFORMIN HYDROCHOLRIDE & SITAGLIPTIN BY USING RP- HPLC

A new method was established for simultaneous estimation of metformin and sitagliptin by RP-HPLC method. The chromatographic conditions were successfully developed for the proportion of metformin and sitagliptin by utilizing SYMMETRYC18 column (4.6×150mm)5µ, flow rate were1ml/min, mobile phase proportion were (70:30 v/v)methanol : phosphate buffer(KH2PO4and K2HPO4) pH 3 ( pH were adjusted with orthophosphoric acid),detection wave length were 258nm. The instrument used were WATERS HPLC Auto Sampler, Sepaproportionn module 2695, photo diode array detector 996, Empower-software version-2. The retention times were found to be 1.694mins and 3.334 mins. The % purity of Metformin and sitagliptin were found to be 100.27% and 99.87% individually. The system suitability parameters for metformin and sitagliptin such as theoretical plates and tailing factor were found to be 2294, 1.27 and 4891 and 1.03, the resolution were found to be 8.67. The analytical method were validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study metformin and sitagliptin were found in consent proportion range of 50µg-250µg and 5µg-50µg and correlation coefficient (r2) were found to be 0.999 and 0.999, % recovery were found to be 99.56% and 99.48%, %RSD for repeatability were 0.27 and 0.40, % RSD for intermediate precision were 0.27 and 0.94individually. The precision study were precise, robust, and repeatable.LOD value were 2.17 and 6.60, and LOQ value were 0.032 and 0.1125individually. Keywords: Metformin , sitagliptin, precise, robust, and repeatable