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

Abstract

A rapid and sensitive method based on liquid chromatography–tandem 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 re-extracted with dichloromethane, after which the analyte and triptorelin as an internal standard (IS) were separated on a 300SB-C18 column (150 mm×4.6 mm i.d., 5 μm 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 multiple-reaction monitoring (MRM) transitions of bivalirudin and IS were at m/z 1091.0→650.4 and m/z 656.5→249.3, respectively. The lower limit of quantification (LLOQ) was 1 ng/mL for 100 μL 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.

Pharmacokinetic study of inosiplex tablets in healthy Chinese volunteers by hyphenated HPLC and tandem MS techniques

“…Although it has been reported [8] , [11] , [12] that PABA is extensively metabolized to p-aminohippuric acid and p-acetaminohippuric acid, for they are not pharmacologically as active as PABA and readily eliminated into the urine, the study here did not emphasize the determination of inosine, but focused on the analysis of DIP and the unconjugated PABA in the biological samples. In this paper, an LC/MS/MS determination method [13] , [14] for DIP and an HPLC method for PABA have been set up, respectively, and the established methods were applied to plasma pharmacokinetic studies of DIP and PABA in 30 healthy Chinese volunteers after oral administration of inosiplex tablets. Meanwhile, the food and gender effects on the pharmacokinetic properties have been investigated as well.…”

The Pharmacokinetics in Mice and Cell Uptake of Thymus Immunosuppressive Pentapeptide Using LC-MS/MS Analysis

“…The addition of 2% FA further improved the peak shape and increased the ionization efficiency of TIPP. However, TIPP is a small peptide with limited solubility in the organic phase, and thus, DCM was supplemented to remove the organic phase [15,16]. Moreover, due to the instability observed between batches, the performance of stabilizers was evaluated, among which DTT had a better stabilizing effect.…”

Development of an LC−MS/MS method for determination of 2-oxo-clopidogrel in human plasma

“…However, the high activity of thiolactone poses great challenge for the establishment of analytic method. For the lucubration of clopidogrel metabolism and individual treatment, this study focused on developing an LC−MS/MS method [18] , [19] to determine the concentration of 2-oxo-clopidogrel. To achieve the goal, the method was validated and applied to a pharmacokinetic study.…”