Direct measurement of active thiol metabolite levels of clopidogrel in human plasma using tris(2‐carboxyethyl)phosphine as a reducing agent by LC–MS/MS. Issue 14 (21st June 2013)

• Record Type: Journal Article
• Authors: Park, Jung Bae; Bae, Soo Hyeon; Jang, Su‐Min; Noh, Won Jun; Hong, Jang‐Hee; Yoon, Kee Dong; Kang, Han Chang; Bae, Soo Kyung
• Is Part Of: Journal of separation science. Volume 36:Issue 14(2013:Jul.)
• Page Start: 2306
• Page End: 2314
• Publication Date: 2013-06-21
• Subjects: Separation (Technology) -- Periodicals; Chromatographic analysis -- Periodicals; 543.089
• DOI: 10.1002/jssc.201300332
• Languages: English
• ISSNs: 1615-9306

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>A simple, robust, and rapid LC–MS/MS method has been 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 C₁₈ RP 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 ESI‐MS. The detector was operated in selected reaction‐monitoring mode at <italic>m/z</italic> 322.0→211.9 for clopidogrel, <italic>m/z</italic> 356.1→155.2 for the AM, and <italic>m/z</italic> 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 (<italic>r</italic>) greater than 0.9976. Precision, both intra‐ and interday, 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.</p> </abstract>