Determination of infliximab in human plasma by liquid chromatography tandem-mass spectrometry
OBJECTIVE To establish a liquid chromatography tandem mass spectrometry(LC-MS/MS)method for the quantification of infliximab in human plasma.METHODS Infliximab was extracted from plasma by methanol.After denatur-ation,reduction and alkylation,trypsin was added to digest infliximab into a large number of peptides.Quantification was per-formed by analyzing signature peptide,and peptide of bevacizumab was used as the internal standard.The chromatographic col-umn was Ultimate LP C18(5 μm,21 mm×100 mm),and the mobile phases were water containing 0.1％ formic acid and acetoni-trile containing 0.1％formic acid.The multiple reaction monitoring was performed in the positive electrospray ionization mode.The established LC-MS/MS method was validated and used to determine plasma drug concentrations of infliximab-treated patients.RESULTS Infliximab showed good correlation within the range of 1-100 μg·mL-1.The intra-and inter-batch accu-racy ranged from 85.0％-115.0％,and the intra-and inter-batch precision was within 15.0％.Specificity,matrix effect,stabil-ity,selectivity and carryover all met the requirements.The plasma concentrations of infliximab in 10 patients with inflammatory bowel disease ranged from 3.61 to 19.60 μg·mL-1.CONCLUSION The LC-MS/MS method established in our study can accu-rately and precisely determine the plasma concentration of infliximab,which provides an effective means for the clinical monitoring of infliximab.
infliximabtherapeutic drug monitoringliquid chromatography tandem-mass spectrometryblood drug concen-tration detection
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