SCVSA Department, University of Parma, Parma, Italy; Laboratory Medicine Service, San Raffaele Hospital, Milano, Italy.
Clinic of Infectious Diseases, San Raffaele Hospital, Milano, Italy.
J Pharm Biomed Anal. 2019 May 30;169:11-18. doi: 10.1016/j.jpba.2019.02.037. Epub 2019 Feb 21.
Antibiotic therapy is a crucial aspect of the management of hospitalized patients, however, current standard dosing protocols have been shown to often attain inadequate plasmatic concentrations which may impair the clinical outcome and promote the selection of multidrug-resistant bacteria. The aim of this study is to establish and validate a robust and fast liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of four commonly used antibiotics (Meropenem, Piperacillin, Linezolid and Teicoplanin) in human plasma according to the European Medicines Agency (EMA) guidelines. Samples preparation was performed using a commercially available extraction kit which needs a very small amount of sample (50 μl). Antibiotics were detected, following a 7 min gradient separation, in multiple reactions monitoring (MRM) mode using a Qtrap 5500 triple quadrupole instrument equipped with an electrospray source operating in positive ion mode. The method, covering the antibiotics' clinically relevant concentration ranges, is also able to quantify, individually, the major teicoplanin components. The high reproducibility and the need of a small amount of sample, associated with the use of a commercial kit, together with a short chromatographic time, makes the method particularly suited for high-throughput routine analysis. Monitoring of plasma antibiotic levels, as part of the clinical routine, would result in a quick therapy adjustment leading to a higher probability of eradicating the infection as well as a potential reduction of multidrug-resistance prevalence. The method was successfully applied to monitor the antibiotic concentration of 49 patients under therapy.
抗生素治疗是住院患者管理的重要方面，然而，目前的标准剂量方案已被证明往往无法达到足够的血浆浓度，这可能会影响临床结果并促进多药耐药菌的选择。本研究的目的是根据欧洲药品管理局（EMA）的指南，建立和验证一种用于同时分析四种常用抗生素（美罗培南、哌拉西林、利奈唑胺和替考拉宁）在人血浆中的灵敏、快速的液相色谱-串联质谱（LC-MS/MS）方法。样品制备采用市售的提取试剂盒，该试剂盒仅需少量样品（50μl）。抗生素在正离子模式下，经 7 分钟梯度分离后，在多重反应监测（MRM）模式下进行检测。该方法涵盖了抗生素的临床相关浓度范围，还能够单独定量替考拉宁的主要成分。该方法具有较高的重现性，需要少量样品，同时还使用了商业试剂盒，再加上较短的色谱时间，使其特别适合高通量常规分析。作为临床常规的一部分，监测血浆抗生素水平将导致快速调整治疗，从而更有可能消除感染，并可能降低多药耐药菌的流行率。该方法已成功应用于监测 49 名接受治疗的患者的抗生素浓度。
