title: Ultrafast quantification of β-lactam antibiotics in human plasma using UPLC-MS/MS.
abstract: There is an increasing interest in monitoring plasma concentrations of β-lactam antibiotics. The objective of this work was to develop and validate a fast ultra-performance liquid chromatographic method with tandem mass spectrometric detection (UPLC-MS/MS) for simultaneous quantification of amoxicillin, cefuroxime, ceftazidime, meropenem and piperacillin with minimal turn around time. Sample clean-up included protein precipitation with acetonitrile containing 5 deuterated internal standards, and subsequent dilution of the supernatant with water after centrifugation. Runtime was only 2.5 min. Chromatographic separation was performed on a Waters Acquity UPLC system using a BEH C18 column (1.7 μm, 100 mm × 2.1 mm) applying a binary gradient elution of water and methanol both containing 0.1% formic acid and 2 mmol/L ammonium acetate on a Water TQD instrument in MRM mode. All compounds were detected in electrospray positive ion mode and could be quantified between 1 and 100 mg/L for amoxicillin and cefuroxime, between 0.5 and 80 mg/L for meropenem and ceftazidime, and between 1 and 150 mg/L for piperacillin. The method was validated in terms of precision, accuracy, linearity, matrix effect and recovery and has been compared to a previously published UPLC-MS/MS method.
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, De Pintelaan 185, Building 2P8, 9000 Ghent, Belgium; Department of Critical Care Medicine, Ghent University, De Pintelaan 185, Building 2K12-IC, 9000 Ghent, Belgium. Electronic address: Mieke.Carlier@ugent.be.
authors.forename: Mieke
authors.initials: M
authors.lastname: Carlier
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, De Pintelaan 185, Building 2P8, 9000 Ghent, Belgium; Department of Laboratory Medicine, Ghent University Hospital, De Pintelaan 185, Building 2P8, 9000 Ghent, Belgium.
authors.forename: Veronique
authors.initials: V
authors.lastname: Stove
authors.affiliation: Department of Critical Care Medicine, Ghent University, De Pintelaan 185, Building 2K12-IC, 9000 Ghent, Belgium.
authors.forename: Jan J
authors.initials: JJ
authors.lastname: De Waele
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, De Pintelaan 185, Building 2P8, 9000 Ghent, Belgium; Department of Laboratory Medicine, Ghent University Hospital, De Pintelaan 185, Building 2P8, 9000 Ghent, Belgium.
authors.forename: Alain G
authors.initials: AG
authors.lastname: Verstraete
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D047090:beta-Lactams / Q000097:blood*
chemical_list: D000900:Anti-Bacterial Agents; D047090:beta-Lactams
keywords: Beta-lactam antibiotics; Therapeutic drug monitoring; UPLC–MS/MS