Abstract
A rapid, sensitive, and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed for the quantitative determination of bivalirudin in human plasma and its application in a pharmacokinetic study. With nafarelin as internal standard, the sample pretreatment involved one-step protein precipitation with methanol of 0.2 mL plasma. The analysis was conducted on Shim-pack XR-ODS II (75 mm × 2.0 mm, i.d., 2.2 µm) column. The mobile phase consisted of acetonitrile and 0.2% formic acid (20:80,v/v) at a flow rate of 0.35 mL min−1. The detection was performed on a triple-quadruple tandem mass spectrometer by multiple reaction monitoring mode via electrospray ionization. The linear calibration curves for bivalirudin were obtained in the concentration range of 1.006–2011 ng mL−1, with a lower limit of quantification of 1.006 ng mL−1. The intra-day and inter-day precision were high, with relative standard deviations lower than 15%. The accuracy in terms of relative error ranged from 2.7% to 7.8% at all quality control levels. The method was successfully applied to a clinical pharmacokinetic study of bivalirudin in healthy volunteers following intravenous administration of bivalirudin.
ACKNOWLEDGMENT
This experiment was supported by the Grant from School of Republic Health and Tropical Medicine of Southern Medical University, China (Grant no. GW201102).