J Korean Soc Clin Pharmacol Ther.  2013 Dec;21(2):166-173. 10.12793/jkscpt.2013.21.2.166.

Validation of LC-MS/MS Method for Determination of Bivalirudin in Human Plasma: Application to a Pharmacokinetic Study

• Kim YH
• Park HJ
• Choi HY
• Lim HS
• Bae KS

• KMID: 2203230
• DOI: http://doi.org/10.12793/jkscpt.2013.21.2.166

Abstract

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 x 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.

Keyword

Bivalirudin; Liquid chromatography; Pharmacokinetics

MeSH Terms

Administration, Intravenous
Angina, Unstable
Chromatography, Liquid
Humans*
Male
Mass Spectrometry
Methods
Nafarelin
Percutaneous Coronary Intervention
Pharmacokinetics
Plasma*
Thrombin
Water
Nafarelin
Thrombin
Water

Figure

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  Figure 1. Chromatograms of (A) a human blank plasma sample; (B) a human plasma sample spiked with 10 ng/mL of bivalirudin and IS (nafarelin); (C) a subject’s plasma sample at 5 min after intravenous bolus administration of bivalirudin 0.375 mg/kg.

• 

  Figure 2. Mean plasma concentration time curves of bivalirudin after single intravenous bolus injection. Data are expressed as mean ± SD (each group, n=6).

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