Abstract
Reduction in the time of analysis whereas maintaining good efficiency has been of substantial focus on high-speed chromatographic separations and UPLC-MS/MS is an analytical instrument for this fast biomedical analysis. In this study a simple, rapid, sensitive and specific UPLC-MS/MS method was developed and validated for quantification of hydrochlorthiazide a common diuretic and anti-hypertensive agent in human plasma. Samples were extracted by protein precipitation using methanol and acetonitrile and on Acquity UPLC BEH (TM) C18 column (50 x 2.1 mm, i.d. 1.7 mu m, Waters, USA) with mobile phase consisted of acetonitrile: 8 mM ammonium acetate: 0.1% formic acid (80: 10: 10 v/v/v) pumped at a flow rate of 0.3 mL/min and detected by tandem mass spectrometry with negative ion mode. The ion transitions recorded in multiple reaction monitoring mode were m/z 295.93 -> 268.90 for hydrochlorthiazide and m/z 427.2 -> 193.08 for internal standard (IS) irbesartan. The assay exhibited a linear dynamic range of 1-500 ng/mL for hydrochlorthiazide in human plasma with good correlation coefficient of (0.997) and with a limit of quantitation of 1 ng/mL. The intra-and inter-assay precisions were satisfactory; the relative standard deviations did not exceed 11.59%. The proposed UPLC-MS/MS method is simple, rapid and highly sensitive, and hence it could be reliable for pharmacokinetic and toxicokinetic study in both animals and humans