Validated LC-MS-MS Method for Quantitative Determination of Batifiban in Human Plasma and Its Application to a Pharmacokinetic Study

Batifiban is a new platelet GPIIb/IIIa receptor antagonist. In this work, an analytical method based on liquid chromatography and electrospray ionization tandem mass spectrometry has been firstly developed and validated for the quantitative measurement of batifiban in human plasma to support the investigation of this compound. Separation of analyte and the internal standard eptifibatide was performed on a Thermo HyPURITY C18 column (150 x 2.1 mm, 5 mu m) with a mobile phase consisting of formic acid 0.1% (v/v)-acetonitrile (40:60, v/v) at a flow rate of 0.25 mL min(-1). The Waters QuattroMicro API triple quadrupole mass spectrometer was operated in multiple reaction monitoring mode via positive electrospray ionization interface using the transition m/z 819.2 -> m/z (623.9 + 159.4) for batifiban and m/z 833.4 -> m/z (645.7 + 159.3) for IS. The method was linear over the concentration range of 2.45-5,000 mu g L-1. The intra- and inter- day precisions were less than 15% in terms of relative standard deviation, and the accuracy was within 8.5% in terms of relative error (RE). The lower limit of quantification (LLOQ) was identifiable and reproducible at 2.45 mu g L-1 with acceptable precision and accuracy. The validated method offered sensitivity and wide linear concentration range. This method was successfully applied for the evaluation of pharmacokinetics of batifiban afer single oral doses of 55, 110 and 220 mu g kg(-1) batifiban to 36 Chinese healthy volunteers.