1. Validation and clinical application of a multiplex high performance liquid chromatography - tandem mass spectrometry assay for the monitoring of plasma concentrations of 12 antibiotics in patients with severe bacterial infections
- Author
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Sabine Laila Lahrichi, Raphaël Burger, Jean-Luc Pagani, O. Marchetti, Nicole Guignard, Thomas Mercier, Nicolas Widmer, Chantal Csajka, Philippe Eggimann, Pascal André, Bertrand Rochat, Eva Choong, Benoît Pesse, Frédéric Lamoth, Sandra Cruchon, Laurent A. Decosterd, Thierry Buclin, and Beatrice Ternon
- Subjects
Adult ,Male ,Clinical Biochemistry ,030226 pharmacology & pharmacy ,01 natural sciences ,Biochemistry ,Meropenem ,Tazobactam ,High-performance liquid chromatography ,Sensitivity and Specificity ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Pharmacokinetics ,Tandem Mass Spectrometry ,medicine ,Humans ,Multiplex ,Chromatography, High Pressure Liquid ,Aged, 80 and over ,ddc:615 ,Chromatography ,medicine.diagnostic_test ,010401 analytical chemistry ,Selected reaction monitoring ,Reproducibility of Results ,Cell Biology ,General Medicine ,Bacterial Infections ,Amoxicillin ,Analysis ,Anti-bacterial agents ,Antibacterial ,Antibiotics ,Assay ,Beta-lactams ,Carbapenems ,Cefepime ,Ceftazidime ,Ceftriaxone ,Cephalosporins ,Cilastatin ,Concentration ,Daptomycin ,Ertapenem ,Flucloxacillin ,Imipenem ,Mass spectrometry ,Penicillins ,Piperacillin ,Plasma ,Quantification ,Rifampicin ,Therapeutic drug monitoring ,0104 chemical sciences ,Anti-Bacterial Agents ,chemistry ,Child, Preschool ,Female ,Drug Monitoring ,medicine.drug - Abstract
Objective Unpredictable pharmacokinetics of antibiotics in patients with life-threatening bacterial infections is associated with drug under- or overdosing. Therapeutic drug monitoring (TDM) may guide dosing adjustment aimed at maximizing antibacterial efficacy and minimizing toxicity. Rapid and accurate analytical methods are key for real-time TDM. Our objective was to develop a robust high-performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) for multiplex quantification of plasma concentrations of 12 antibiotics: imipenem/cilastatin, meropenem, ertapenem, cefepime, ceftazidime, ceftriaxone, piperacillin/tazobactam, amoxicillin, flucloxacillin, rifampicin, daptomycin. Methods A single extraction procedure consisting in methanol plasma protein precipitation and H2O dilution was used for all analytes. After chromatographic separation on an Acquity UPLC HSS-T3 2.1 × 50 mm, 1.8 µm (Waters®) column, quantification was performed by electro-spray ionisation-triple quadrupole mass spectrometry with selected reaction monitoring detection. Antibiotics were divided in two pools of calibration according to the frequency of analyses requests in the hospital routine antibiotic TDM program. Stable isotopically-labelled analogues were used as internal standards. A single analytical run lasted less than 9 min. Results The method was validated based on FDA recommendations, including assessment of extraction yield (96–113.8%), matrix effects, and analytical recovery (86.3–99.6%). The method was sensitive (lower limits of quantification 0.02–0.5 µg/mL), accurate (intra/inter-assay bias −11.3 to +12.7%) and precise (intra/inter-assay CVs 2.1–11.5%) over the clinically relevant plasma concentration ranges (upper limits of quantification 20–160 µg/mL). The application of the TDM assay was illustrated with clinical cases that highlight the impact on patients’ management of an analytical assay providing information with short turn-around time on antibiotic plasma concentration. Conclusion This simple, robust high-throughput multiplex HPLC-MS/MS assay for simultaneous quantification of plasma concentrations of 12 daily used antibiotics is optimally suited for clinically efficient real-time TDM.
- Published
- 2020