Quantitation of bivalirudin, a novel anticoagulant peptide, in human plasma by LC-MS/MS： Method development, validation and application to pharmacokinetics

• Published in: Journal of pharmaceutical analysis Vol. 3; no. 1; pp. 1 - 8
• Main Authors: Li, Xiao-Jiao, Sun, Yan-Tong, Yin, Lei, Zhang, Xue-Ju, Yang, Yan, Paul Fawcett, J., Cui, Yi-Min, Gu, Jing-Kai
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.02.2013; Research Center for Drug Metabolism, College of Life Science, Jilin University, Changchun 130012, PR China%Clinical Pharmacology Center, Research Institute of Translational Medicine, The First Bethune Hospital of Jilin University,Changchun 130061, PR China%Research Center for Drug Metabolism, College of Life Science, Jilin University, Changchun 130012, PR China%School of Pharmacy, University of Otago, P.O.Box 56, Dunedin, New Zealand%Department of Pharmacy, Peking University First Hospital, Beijing 100034, PR China; Clinical Pharmacology Center, Research Institute of Translational Medicine, The First Bethune Hospital of Jilin University,Changchun 130061, PR China; Xi'an Jiaotong University
• Subjects: Anticoagulant; Bivalirudin; Human plasma; LC-MS; LC–MS/MS; Pharmacokinetics; S方法; 人体血浆; 应用; 开发; 抗凝血剂; 药代动力学; 验证; Bivalirudin; Anticoagulant; Human plasma; Pharmacokinetics; LC–MS/MS; LC-MS/MS
• ISSN: 2095-1779; 2214-0883
• DOI: 10.1016/j.jpha.2012.10.006

A rapid and sensitive method based on liquid chromatographtandem mass spectrometry （LC-MS/MS） for the determination of a novel anticoagulant peptide bivalirudin in human plasma has been developed and validated. Plasma samples were precipitated protein with acetonitrile and reextracted with dichloromethane, after which the analyte and triptorelin as an internal standard （IS） were separated on a 300SB-Cl8 column （150 mm x 4.6 mm i.d., 5 gm particle size） using 0.1% formic acid：methanol （45：55, v/v） as mobile phase. The triple-quadrupole mass spectrometer, equipped with electrospray ionization （ESI） interface, was operated in the positive ion mode, and the multiplereaction monitoring （MRM） transitions of bivalirudin and IS were at m/z 1091.0-650.4 and m/z656.5 - 249.3, respectively. The lower limit of quantification （LLOQ） was 1 ng/mL for 100 ng/mL plasma sample and the assay was linear over the concentration range 1 1000 ng/mL. The accuracy was within a range from -0.4% to 0.5% in terms of relative error （RE） and the intra- and inter-day precisions in terms of relative standard deviation （RSD） were 〈2.92 and 〈 3.36, respectively. The method was successfully applied to a pharmacokinetic study involving intravenous administration of bivalirudin （0.5 mg/kg） to Chinese volunteers.