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

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.

Species Comparison of Pre-systemic Bioactivation of Vicagrel, a New Acetate Derivative of Clopidogrel

“…The harvested plasma was processed by the addition of triple volume of cold acetonitrile (containing 50 ng/mL ISTD, 5% acetic acid and 10 mM MPBr) to avoid the possible degradation caused by plasma proteins and derivatize AM ( Qiu et al, 2013 ). Also, 1,4-dithio- DL -threitol (DTT, as antioxidant) significantly increased the stability of 2-oxo-clopidogrel ( Li et al, 2015 ). In our study, the acidification by acetic acid could also prevent the degradation of 2-oxo-clopidogrel (from below 50% to above 85%) by compared to the samples without acetic acid treatment.…”