Determination of batifeiban in macaque plasma by LC-MS/MS and its pharmacokinetic study

Objective: To develop a LC-MS/MS method for determination of batifeiban in macaque plasma,and to investigate the pharmacokinetics of batifeiban.Methods: Plasma samples 200 μL were treated with 600 μL mixture of acetonitrile and methanol(70∶30,containing 2 μg·mL-1 of eptifibatide).After centrifugation to precipitate the proteins,the supernatant was evaporated under nitrogen gas flow at 40 ℃.Residues were dissolved in 100 μL of mobile phase and centrifuged at 12 000 r·min-1 for 5 min.Thereafter,20 μL aliquots were injected into the LC-MS/MS for analysis.Separation was performed on a Diamonsil C18 column(150 mm×4.6 mm,5 μm).The mobile phase was composed of methanol and 20 mmol·L-1 ammonium formate(50∶50).The selected reaction monitoring(SRM) transitions were performed at m/z 818.3→632.4 for batifiban and m/z 832.0→646.2 for eptifibatide(IS) in the positive ion mode with electrospray ionization(ESI) source.Results: Calibration curve was linear over the concentration range of 25~5 000 ng·mL-1.The lower limit of quantification was 25 ng·mL-1.The mean recovery was(65.7±4.9)%.The intra-and inter-day precisions were less than 5.3% and 14.0%,respectively,and the accuracy was within ±7.5%.Batifiban was stable in possible conditions of storing and handling.Conclusion: The method has been validated,and can be applied to determine the plasma concentration of batifiban for a pharmacokinetic study.