Simultaneous estimation of saxagliptin and dapagliflozin in human plasma by validated HPLC-UV method

Abstract

Objective: The fixed dose combination of saxagliptin and dapagliflozin, recently approved antidiabetic medication. It is marketed with a brand name Qtern. The intend method aim to develop a simple, rapid, sensitive and validated isocratic reversed phase high performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of saxagliptin and dapagliflozin in human plasma by using linagliptin as internal standard as per US-FDA guidelines. Materials and methods: The method was performed on Waters 2695 HPLC equipped with quaternary pump. The analyte separation was achieved using Eclipse XDB C18 (150×4.6μ×5mm) column with a mobile phase consisting of 0.1% ortho phosphoric acid and acetonitrile (50:50) with pH adjusted to 5.0 at1ml/min flow rate. Results: The analyte was detected at 254nm. Retention time of the internal standard, saxagliptin and dapagliflozin was found at 2.746, 5.173 and 7.218minutes, respectively. The peaks were found to be free of interference. The method is validated over a dynamic linear range of 0.01 to 0.5μg/ml and 0.05 to 2μg/ml for saxagliptin and dapagliflozin, respectively, with a correlation coefficient of 0.998. The precision and accuracy of samples of six replicate measurements at LLOQ level was within limit. The analytes were found to be stable in human plasma at -28°C for 37 days. Conclusion: The stability, sensitivity, specificity and reproducibility of this method make it appropriate for the determination of saxagliptin and dapagliflozin in human plasma.