UPLC–MS/MS determination of thiamphenicol in human plasma and its application to a pharmacokinetic study

Abstract A sensitive and rapid ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed to determine thiamphenicol (TAP) in human plasma using chlorzoxazone as the internal standard (IS). Sample preparation was accomplished through a liquid–liquid extraction procedure with ethyl acetate to precipitation of plasma protein, and to a 0.1 mL plasma sample. The analyte and IS were separated on an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with the mobile phase of acetonitrile and 1% formic acid in water with gradient elution at a flow rate of 0.40 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) by multiple reactions monitoring (MRM) of the transitions at m / z 354.3 → 185.1 for TAP and m / z 168.1 → 132.1 for IS. The linearity of this method was found to be within the concentration range of 10–8000 ng/mL with a lower limit of quantification of 10 ng/mL. Only 1.5 min was needed for an analytical run. The method herein described was superior to previous methods and was successfully applied to the pharmacokinetic study of TAP in healthy Chinese volunteers after oral administration.