QUANTITATIVE DETERMINATION OF BIVALIRUDIN IN HUMAN PLASMA BY ULTRA-FAST LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY AND ITS APPLICATION IN A PHARMACOKINETIC STUDY

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.

Keywords:

• bivalirudin
• human plasma
• nafarelin
• pharmacokinetic
• protein precipitation
• UFLC-ESI-MS/MS

ACKNOWLEDGMENT

This experiment was supported by the Grant from School of Republic Health and Tropical Medicine of Southern Medical University, China (Grant no. GW201102).