A validated LC-MS/MS bioanalytical method for the simultaneous determination of dapagliflozin or saxagliptin with metformin in human plasma

An LC-MS/MS method was developed and validated for the simultaneous quantitation of dapagliflozin or saxagliptin with metformin in human plasma. Separation and detection of dapagliflozin (DAP), saxagliptin (SAX), metformin (MET) and their internal standards was achieved on Zorbax C18 column (50 × 4.6 mm, 5 μm) using acetonitrile: 0.1% formic acid (45:55, v/v) as a mobile phase applying electrospray ionization (ESI) source in positive ion mode. Sequential liquid-liquid extraction and precipitation techniques were used for the extraction due to the difference in the physicochemical properties of drugs. The MS/MS response was linear over the concentration ranges from 5 to 500 ng/ml, 2–50 ng/ml and 10–4000 ng/ml for DAP, SAX and MET, respectively. Precision, accuracy, selectivity, recovery, and stability were within the acceptable limits as FDA guidelines. The method is suitable for further application of pharmacokinetic studies. •The prevalence of T2DM indicates the use of combination therapy such as combination between Gliflozins and Gliptins with metformin.•Rapid LC-MS/MS method was developed for the determination of dapagliflozin or saxagliptin with metformin.•Separation and determination of the three antidiabetic drugs were performed in human plasma.•Method was validated according to FDA and EMA guidelines.