Rapid determination of canagliflozin in rat plasma by UHPLC–MS/MS using negative ionization mode to avoid adduct-ions formation

•A UHPLC–MS/MS assay for determination of canagliflozin in plasma.•Use of negative ionization mode to avoid adduct-ions formation.•Assay applied in pre-clinical pharmacokinetic study in rats.

Canagliflozin is the first sodium–glucose co-transporter-2 inhibitor, approved by the US Food and Drug Administration for the treatment of type 2 diabetes mellitus. In this study, a sensitive UHPLC–MS/MS assay for rapid determination of canagliflozin in rat plasma was developed and validated for the first time. Chromatographic separation of canagliflozin and zafirlukast (IS) was carried out on Acquity BEH C18 column (100×2.1 mm, i.d. 1.7 µm) using acetonitrile–water (80:20, v/v) as mobile phase at a flow rate of 0.3 mL min−1. Canagliflozin and IS were extracted from plasma by protein precipitation method using acetonitrile. The mass spectrometric detection was performed using electrospray ionization source in negative mode to avoid canagliflozin adduct ions formation. Multiple reaction monitoring were used for quantitation of precursor to product ion at m/z 443.16 >364.96 for canagliflozin and m/z 574.11>462.07 for IS, respectively. The assay was fully validated in terms of selectivity, linearity, accuracy, precision, recovery, matrix effects and stability. The validated method was successfully applied to the characterization of oral pharmacokinetic profiles of canagliflozin in rats. The mean maximum plasma concentration of canagliflozin of 1616.79 ng mL−1 was achieved in 1.5 h after oral administration of 20 mg kg−1 in rats.

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