A simple, precise and specific reverse phase high performance liquid chromatographic method has been developed and validated for the determination of sitagliptin in blood. The HPLC separation was carried out by reverse phase chromatography on Shimadzu HPLC system consisted of welchrom 5µ C18 column (250 X 4.6 mm), SPD 10A UV detector and LC 10AD Pumps. Rheodyne injector with 20 μl capacity, mobile phase comprises of Methanol: Water (65:35) pH 9.0 adjusted with triethanolamine at flow rate 1.0 ml/min. Glimperide was used as internal standard in the determination. The retention time of sitagliptin and internal standard was found to be 5.73 and 12.2 respectively. The detection was monitored at 262 nm. The calibration curve for sitagliptin was linear from 0.1-10 µg/ml with correlation coefficient of 0.999. The inter-day and intraday precision was found to be within limits. The method was validated as per the guidelines.
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