BACKGROUND The accurate measurement of plasma levels of antibiotics is crucial for the individualization of antimicrobial therapies based on PK/PD strategies. In this paper we describe a new rapid and simple LC-MS/MS platform for quantifying 14 antibiotics (amikacin, amoxicillin, ceftazidime, ciprofloxacin, colistin, daptomycin, gentamicin, linezolid, meropenem, piperacillin, teicoplanin, tigecycline, tobramycin and vancomycin) and a beta-lactamase inhibitor (tazobactam) starting from 50 μL plasma samples. METHODS Analyses were performed on a Thermo Scientific™ Ultimate™ 3000 LC system (Thermo Fisher Scientific, Milan, Italy) coupled to a Thermo Scientific™ TSQ Quantiva™ Triple Quadrupole mass spectrometer. After fast protein precipitation protocols and addition of deuterated internal standards, samples were subjected to a fast HPLC gradient separation and the 15 drugs were quantified using multiple reaction monitoring of specific transitions over a wide range of concentrations. The suitability of the assay for TDM was tested on plasma samples derived from pediatric patients under treatment with one or more antibiotics. RESULTS The overall turnaround time of the assay was 20 min. The assay was validated following EMA guidelines for bioanalytical method validation and showed excellent accuracy (ranging from 85.3 and 112.7) and reproducibility (ranging from 1.3 to 9.7) as well as the absence of matrix effects (<15 %) for all the drugs tested. The lower limits of quantifications were between 0.1 and 2 mg/L. the recovery rate exceeded 85 % for all the drug tested. Stability was evaluated in different conditions thus allowing the setting up of reliable operative procedures. CONCLUSIONS This work provides a LC-MS/MS platform validated for clinical use for a rapid quantification of a broad spectrum of drugs having different chemical characteristics in a small volume of plasma and is suitable for real-time TDM-guided personalization of antimicrobial treatment in critically ill patients.

中文翻译：

---

用于14种抗生素的常规治疗药物监测的液相色谱-串联质谱分析平台：适用于危重儿科患者。

背景技术准确地测量抗生素的血浆水平对于基于PK / PD策略的抗微生物疗法的个体化至关重要。在本文中，我们描述了一种新的快速简便的LC-MS / MS平台，用于定量14种抗生素（阿米卡星，阿莫西林，头孢他啶，环丙沙星，粘菌素，达托霉素，庆大霉素，利奈唑胺，美罗培南，哌拉西林，替考拉宁，替加环素，妥布霉素和万古霉素）从50μL血浆样品开始的β-内酰胺酶抑制剂（他唑巴坦）。方法在与Thermo Scientific™TSQ Quantiva™三重四极杆质谱仪耦合的Thermo Scientific™Ultimate™3000液相色谱系统（Thermo Fisher Scientific，意大利米兰）上进行分析。经过快速蛋白质沉淀方案并添加氘化内标后，对样品进行快速HPLC梯度分离，并使用多种反应监测多种浓度范围内的特定跃迁对15种药物进行定量。在接受一种或多种抗生素治疗的儿科患者的血浆样品上测试了TDM分析的适用性。结果分析的总周转时间为20分钟。该测定法按照EMA指导原则进行了生物分析方法验证，并显示出极好的准确性（范围从85.3和112.7）和可重复性（范围从1.3到9.7），并且对于所有测试的药物均无基质作用（<15％）。定量的下限在0.1和2 mg / L之间。所有测试药物的回收率均超过85％。在不同条件下评估稳定性，从而建立可靠的手术程序。结论这项工作提供了一种LC-MS / MS平台，该平台经验证可用于临床，可在小剂量血浆中快速定量分析具有不同化学特性的多种药物，并且适用于TDM实时指导的抗菌治疗个性化危重病人。