Development and validation of an LC-ESI-MS/MS method for the quantitation of hemslecin A in rhesus monkey plasma and its application in pharmacokinetics

In this study, a new LC-ESI-MS/MS-based method was validated for the quantitation of hemslecin A in rhesus monkey plasma using otophylloside A as internal standard (IS). Hemslecin A and the IS were extracted from rhesus monkey plasma using liquid-liquid extraction as the sample clean-up procedure, and were subjected to chromatography on a Phenomenex Luna CN column (150x2.0mm, 3.0 mu m) with the mobile phase consisting of methanol and 0.02mol/mL ammonium acetate (55:45, v/v) at a flow rate of 0.2mL/min. Detection was performed on an Agilent G6410B tandem mass spectrometer by positive ion electrospray ionization in multiple reaction monitoring mode, monitoring the transitions m/z 580.5 [M+NH4](+)503.4 and m/z 518.2 [M+NH4](+)345.0 for hemslecin A and IS, respectively. The assay was linear over the concentration range of 0.5-200ng/mL and was successfully applied to a pharmacokinetic study in rhesus monkeys. Copyright (c) 2013 John Wiley & Sons, Ltd.