Simultaneous Determination of Canagliflozin and Metformin in Human Plasma by LC–MS/MS Assay and its Application to a Human Pharmacokinetic Study

Abstract:

Objective: The main objective of this work was to develop a simple, rapid and sensitive liquid chromatography/tandem mass spectrometry (LC–MS/MS) method for the simultaneous quantification of Canagliflozin and Metformin. Methods: Deuterated compounds of respective drugs were used an internal standard. Sample extraction was carried out using a simple Protein Precipitation (PP) technique. A C₁₈ column with an isocratic mobile phase composed of 5mM ammonium acetate with 0.01% formic acid and methanol were used for chromatographic separation. Results: The method was validated in the linearity range of 10.00–6028.00 ng/mL for Canagliflozin and 10.00– 3027.00 ng/mL for Metformin. The precision and accuracy results over five concentration levels in five different batches were well within the acceptance limits. A variety of stability tests were executed in plasma and in neat samples are comply with the FDA guidelines. Conclusion: The proposed assay method is simple, rapid and sensitive for the simultaneous determination of Canagliflozin and Metformin in human plasma. This method was successfully used to quantitate the in vivo plasma concentrations obtained from a pharmacokinetic study.

Key words: Canagliflozin, Metformin, Human Plasma, LC–MS/MS, Method Validation, Pharmacokinetics.