Validation of LC-MS/MS Method for Determination of Bivalirudin in Human Plasma: Application to a Pharmacokinetic Study
Background: Bivalirudin is a direct thrombin inhibitor for patients with unstable angina undergoing
percutaneous coronary intervention.
Methods: A sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) method was
developed and validated for the determination of bivalirudin, in human plasma using nafarelin as internal
standard (IS). Chromatographic separation was performed using a Shiseido MG3 mm column (2.0 × 50
mm) with a gradient mobile phase consisting of water and acetonitrile containing 0.1 % formic acid at
a flow rate of 0.4 mL/min, and total run time was within 5 min. Detection and quantification was
performed by the mass spectrometer using a multiple reaction-monitoring mode at m/z 1091.0 → 650.3
for bivalirudin, and m/z 662.1 → 249.3 for IS.
Results: The assay was linear over a concentration range of 10 - 10000 ng/mL with a lower limit of
quantification of 10 ng/mL in human plasma.
Conclusion: This method was successfully applied for pharmacokinetics study after intravenous
administration of bivalirudin to healthy Korean male volunteers.