Abstract
With the objective of decreasing analysis time and retaining good efficiency (UPLC-MS/MS) is an outstanding analytical approach for speedy biomedical analysis. The aim of this study was to develop and validate a simple, rapid, sensitive and specific UPLC-MS/MS method for quantification of CZT in human plasma. After a simple protein precipitation using acetonitrile and methanol, CZT and paroxetine (IS) were separated on Acquity UPLC BEH (TM) C-18 column (50 x 2.1 mm i.d., 1.7 mu m, Waters, USA) using a mobile phase composed of methanol:0.1%(v/v) ammonium hydroxide (80:20) pumped at a flow rate of 0.4 mL min(-1). CZT and IS were eluted at 0.46 and 0.66 min, respectively. The mass spectrometric determination was carried out using an electrospray interface operated in the positive mode with multiple reaction monitoring mode. The mass transitions m/z 450.0 -> 260.0 and m/z 330.11 -> 192.11 were used to measure CZT and internal standard, respectively. Mass transition m/z 450.0 -> 176.99 was used as qualifying ion for CZT. The method was linear in the concentration range of 5-500 ng mL(-1) with correlation coefficient of 0.997 and lower limit of quantitation of 5 ng mL(-1). This study represents the first report describing the determination of CZT in human plasma by UPLC-MS/MS method. The proposed method is superior to the previously reported LC-MS methods in terms of the sensitivity and simplicity as the method described herein is based on simple one step protein precipitation for sample preparation and isocratic flow of mobile phase. The run time was only 2 min which is suitable for high-throughput analysis.