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. 2015 Feb;5(1):12-17.
doi: 10.1016/j.jpha.2014.07.004. Epub 2014 Jul 17.

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

Yu-Han Li  1 , Min Song  1 , Tai-Jun Hang  1
Affiliations

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

Yu-Han Li et al. J Pharm Anal. 2015 Feb.

Abstract

A sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was established to determine 2-oxo-clopidogrel, a crucial intermediate metabolite in human plasma. A chromatographic separation was performed on a Sapphire C18 column following a liquid-liquid extraction sample preparation with methyl t-butyl ether. Detection was carried out on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) with an electrospray ionization (ESI) mode. The method was validated in terms of specificity, accuracy, precision and limit of quantification. The calibration curves ranged from 0.50 to 50.0 ng/mL with good linearity. The stability was fully validated with addition of 1,4-dithio-DL-threitol (DTT) into the plasma sample prior to and in the preparation procedure. The validated method was proved to be suitable for use in pharmacokinetic study after single oral administration of 75 mg clopidogrel tablets in human subjects, which could make contribution to intensive study of the clinical drug-drug interactions of clopidogrel and individual treatment.

Keywords: 2-Oxo-clopidogrel; Human plasma; LC−MS/MS; Pharmacokinetics.

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Figures

Fig. 1
Fig. 1
Active metabolic pathway of clopidogrel.
Fig. 2
Fig. 2
Product ion spectra of (A) 2-oxo-clopidogrel and (B) IS.
Fig. 3
Fig. 3
LC−MS/MS of (A) Blank plasma extract, (B) Spiked LLOQ (0.5 ng/mL, tR=3.8 min) and mifepristone (1 μg/mL, tR=5.0 min), and (C) Plasma sample collected from a subject at 2 h elapsed from administration of 75 mg clopidogrel(6.50 ng/mL 2-oxo-clopidogrel and 1 μg/mL mifepristone).
Fig. 4
Fig. 4
Plasma concentration-time profile (mean±SD, n=8) of 2-oxo-clopidogrel after an administration of 75 mg Plavix in human subjects.
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