A Simple and Sensitive LC–MS/MS Method for Determination of Dapagliflozin and Its Major Metabolite in Human Plasma

Dapagliflozin (DAPA), an inhibitor of sodium-dependent glucose cotransporter-2, has been created to treat individuals with type 2 diabetes mellitus. A method was developed and validated using high-performance liquid chromatography coupled to tandem mass spectrometry to aid in studying the connection between clinical effectiveness and concentration of DAPA and its primary metabolite dapagliflozin 3-O-glucuronide (D3OG) in human plasma. The two analytes were separated using the Waters XSELECT HSS T3 (2.1 × 100 mm, 3.5 μm; Waters Co., Milford, USA) chromatographic column, with a mobile phase flow rate of 0.3 mL/min. The elution program was performed after protein precipitation with methanol, which only required a 5-min duration. The extraction recovery was from 99.8 to 109% for DAPA and from 101 to 103% for D3OG. Validation of the method for detecting DAPA within the range of 5-50 ng/mL and D3OG within the range of 50-500 ng/mL demonstrated satisfactory inter- and intra-day precision and accuracy. The method was successfully developed and validated, and it was used to measure the levels of DAPA and D3OG in plasma samples from patients with type 2 diabetes mellitus.