A simple and sensitive high performance liquid chromatography assay with a fluorescence detector for determination of canagliflozin in human plasma

Abstract

Canagliflozin is the first sodium–glucose co-transporter-2 inhibitor approved for the treatment of type 2 diabetes mellitus. In this study, a simple and sensitive HPLC assay with a florescence detector was developed for accurate quantification of canagliflozin in human plasma using telmisartan as the internal standard (IS). Plasma samples were extracted by a liquid–liquid extraction method using diethyl ether as an extracting solvent. Chromatographic separation of canagliflozin and IS was performed on a Nucleodur Isis C₁₈ column with an isocratic mobile phase of 20 mM potassium dihydrogen orthophosphate : acetonitrile (45 : 55, v/v) at a flow rate of 1 mL min⁻¹. Canagliflozin and IS were eluted at 2.8 and 5.8 min, respectively, and detected at 280 and 325 nm for excitation and emission, respectively. The plasma calibration curve displayed excellent linearity over the concentration range of 16.13–6000 ng mL⁻¹. The assay was fully validated in terms of selectivity & specificity, linearity of the calibration curve, accuracy & precision, recovery and stability under various storage conditions. To the best of our knowledge, this is the first validated HPLC-florescence detector assay for the quantification of canagliflozin in human plasma.