Direct Measurement Of Active Thiol Metabolite Levels Of Clopidogrel In Human Plasma Using Tris (2-carboyxethyl) Phosphine As A Reducing Agent By LC-MS/MS | 10301
Journal of Analytical & Bioanalytical Techniques
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A simple, robust, and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed
and validated for the simultaneous quantitation of clopidogrel and its active metabolite (AM) in human plasma. Tris
(2-carboxyethyl) phosphine (TCEP) was used as a reducing agent to detect the AM as a disulfide bonded complex with plasma
proteins. Mixtures of TCEP and human plasma were deproteinized with acetonitrile containing 10 ng/mL of clopidogrel-d4 as an
internal standard (IS). The mixtures were separated on a C
reversed-phase column with an isocratic mobile phase consisting of
0.1% formic acid in acetonitrile and water (90:10, v/v) at a flow rate of 0.3 mL/min. Detection and quantification were performed
using a mass spectrometer with a positive electrospray ionization source. The detector was operating in selected reaction-
monitoring mode at m/z 322.0→211.9 for clopidogrel, m/z 356.1→155.2 for the AM, and m/z 326.0→216.0 for the IS. The linear
dynamic range for clopidogrel and its AM were 0.05?20 and 0.5?200 ng/mL, respectively, with correlation coefficients (r) greater
than 0.9976. Precision, both intra- and inter-day, was less than 8.26% with an accuracy of 87.6?106%. The validated method was
successfully applied to simultaneously analyze clinical samples for clopidogrel and its AM.
Jung Bae Park is a Ph.D. candidate in drug metabolism and pharmacokinetics in the Catholic University of Korea. He graduated from the Catholic
University of Korea in 2013, where he majored in pharmacokinetics.
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