El-Najjar Nahed, Hösl Julian, Holzmann Thomas, Jantsch Jonathan, Gessner André
Institute of Clinical Microbiology and Hygiene, University Hospital Regensburg, Germany.
Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Germany.
Drug Test Anal. 2018 Mar;10(3):584-591. doi: 10.1002/dta.2253. Epub 2017 Aug 23.
A large variation in the levels of different ß-lactams and other antibiotics used in critically ill patients has been documented. The aim of this study is to establish and validate a fast, ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous analysis of ten antibiotics (Meropenem, Cefepime, Ceftazidime, Piperacillin, Benzylpenicillin, Ampicillin, Flucloxacillin, Linezolid, and Sulfamethoxazole/Trimethoprim) in human plasma according to European Medicines Agency (EMA) guidelines. Protein precipitation with ice-cold methanol containing 9 isotopically labeled internal standards was used for sample clean up. Antibiotics were detected, following a 4-minute gradient separation, in multiple reactions monitoring (MRM) using API 4000 instrument equipped with electrospray source operating in positive ion mode. The lower limit of quantification was 0.1 mg/L for Meropenem, Ceftazidime, Piperacillin, Ampicillin, Flucloxacillin, and Sulfamethoxazole; 0.05 mg/L for Cefepime, Benzylpenicillin, and Trimethoprim; and 0.02 mg/L for Linezolid. The method proved to be precise and accurate and applicable for therapeutic drug monitoring and other pharmacokinetic studies.
据记录，危重症患者使用的不同β-内酰胺类抗生素和其他抗生素的水平存在很大差异。本研究的目的是根据欧洲药品管理局（EMA）指南，建立并验证一种快速、超高效液相色谱-串联质谱（UPLC-MS/MS）方法，用于同时分析人血浆中的十种抗生素（美罗培南、头孢吡肟、头孢他啶、哌拉西林、苄青霉素、氨苄西林、氟氯西林、利奈唑胺和磺胺甲恶唑/甲氧苄啶）。采用含9种同位素标记内标的冰冷甲醇进行蛋白沉淀以净化样品。使用配备电喷雾源且以正离子模式运行的API 4000仪器，在多反应监测（MRM）中经过4分钟梯度分离后检测抗生素。美罗培南、头孢他啶、哌拉西林、氨苄西林、氟氯西林和磺胺甲恶唑的定量下限为0.1 mg/L；头孢吡肟、苄青霉素和甲氧苄啶为0.05 mg/L；利奈唑胺为0.02 mg/L。该方法被证明是精确、准确的，适用于治疗药物监测和其他药代动力学研究。
