UPLC for the Determination of Clopidogrel in Dog Plasma by Tandem Quadrupole MS: Application to a Pharmacokinetic Study

A rapid, sensitive and specific ultra performance liquid chromatography-electrospray ionization tandem quadrupole mass spectrometry method was developed and validated for the determination of clopidogrel in dog plasma. Plasma samples were deproteinized with acetonitrile and separated on a Waters BEH C18 column (1.7 μm, 50 mm × 2.1 mm id.) with isocratic elution at a flow-rate of 0.2 mL min^?1 and mobile phase consisting of acetonitrile and water (containing 0.15% formic acid) (75:25, v/v). The single run analysis was as shorter as 2 min. Electrospray ionization in positive ion mode and multiple reaction monitoring were used for the quantification of clopidogrel with monitored transitions m/z 322 → 212 for clopidogrel and m/z 324 → 217 for internal standard (gliclazide). The intra- and inter-day precisions (RSD%) were less than 6.32 and 7.03%, and accuracy (RE%) between ?9.12 and 9.65% (n = 6). The extraction recovery of clopidogrel was 96.7%. The developed method was successfully applied to the pharmacokinetic study of clopidogrel tablets in dogs following oral administration at a single dose of 75 mg.