Development and validation of a LC-MS/MS method for quantification of dapaglifozin in human plasma

A specific and sensitive LC-MS/MS method has been developed for determination of dapaglifozin (DAP) in human plasma. DAP and the internal standard (empaglifozin) were extracted from human plasma by liquid-liquid extraction. Chromatographic separation was executed on a Hypersil GOLD C18 column (50 x 2.1 mm, 1.7 µm) with gradient elution at a fow rate of 0.3 ml/min using a mixture of water and acetonitrile as mobile phase. The MS/MS detection of dapagliflozin and IS was done in the multi reaction-monitoring (MRM) mode using ESI ion source at negative mode, with quantitative transitions at (m/z) 407.27 →329.19 for DAP, 449.29 → 371.18 for (IS). The method was validated according to current requirements of USFDA guideline over concentration range from 2.50 ng/mL to 250.0 ng/mL. The recovery range in accuracy study were between 90.8% and 103.6%. The intra-day and inter-day precisions of the method, explained in CV(%), were not more than 6.2%. This method was proved as suitable for bioanalysis in applications such as bioequivalent (BE) studies.