Simultaneous quantification of multiple anti-diabetic drugs in whole blood by ultra-performance liquid chromatography-tandem mass spectrometry

Abstract

An analytical method was developed and validated for the simultaneous quantification of multiple anti-diabetic drugs in whole blood using ultra-performance liquid chromatography-tandem mass spectrometry. Chromatographic separation was achieved on a CAPCELL PAK INERT ADME-HR column. The mobile phases consisted of water containing 0.1% formic acid and acetonitrile containing 0.1% formic acid. Quantification of 10 antidiabetic drugs was performed by selected reaction monitoring with each product ion referenced against 4 isotopically labelled internal standards. Calibration curves exhibited good linear relationships ranging from 0.0002–20 μg/mL, with correlation coefficients exceeding 0.996. The limits of detection were estimated to be 0.0001–0.005 μg/mL. The intra- and inter-day accuracies and precisions were 98.6–105.5% and 0.1–8.0%, respectively. The recovery efficiencies were in the range of 74.4–102.3%. Matrix effects were observed ranging from -29.1 to 534.6%. Stability tests of the analytes revealed that gliclazide was relatively unstable when stored long-term (28 days) at −30°C. We expect that the method described in the present study will be useful for various studies such as clinical and forensic toxicological investigations.