Development and validation of an LC–MS/MS method for the determination of tofogliflozin in plasma and its application to a pharmacokinetic study in rats

•A rapid and sensitive LC–MS/MS method was developed for tofogliflozin quantification in rat plasma.•The method has a low quantification limit of 0.5 ng/mL and short run time of 4 min.•Plasma concentration was quantified at least 24 h after oral administration of 0.4 mg/kg tofogliflozin to rats.

Tofogliflozin is a novel selective inhibitor of sodium-dependent glucose co-transporter-2 (SGLT2) and has been developed for the treatment of patients with type 2 diabetes mellitus. In this study, a highly sensitive and specific liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for the quantitation of tofogliflozin in rat plasma was developed and validated. The detection was performed using an API 3200 triple-quadrupole mass spectrometer with selected reaction monitoring (SRM) in the positive electrospray ionization mode. The SRM transitions were m/z = 387.1 [M+H]+ → 267.1 for tofogliflozin and m/z = 451.2 [M+H]+ → 71.0 for empagliflozin (internal standard: I.S.). Chromatographic separation was performed on a Quicksorb ODS (2.1 mm i.d. × 150 mm, 5 μm size) using isocratic elution with acetonitrile/10 mM ammonium acetate (50:50, v/v) as the mobile phase at a flow rate of 0.2 mL/min and the total run time was 4.0 min. The lower limit of quantification (LLOQ) for tofogliflozin was 0.5 ng/mL with sufficient specificity, accuracy, and precision. The validated method was successfully applied to the pharmacokinetic studies of tofogliflozin in rats. This assay method could be a valuable tool for future studies including pharmacokinetic and pharmacodynamic studies of SGLT2 inhibitors.