Cizmarova Ivana, Mikus Peter, Svidrnoch Martin, Piestansky Juraj
Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius University in Bratislava, Odbojarov 10, SK-832 32 Bratislava, Slovakia.
Toxicological and Antidoping Center, Faculty of Pharmacy, Comenius University in Bratislava, Odbojarov 10, SK-832 32 Bratislava, Slovakia.
Pharmaceuticals (Basel). 2024 Apr 19;17(4):526. doi: 10.3390/ph17040526.
Monitoring plasma concentrations of β-lactam antibiotics is crucial, particularly in critically ill patients, where variations in concentrations can lead to treatment failure or adverse events. Standardized antimicrobial regimens may not be effective for all patients, especially in special groups with altered physiological parameters. Pharmacokinetic/pharmacodynamic (PK/PD) studies highlight the time-dependent antibacterial activity of these antibiotics, emphasizing the need for personalized dosing. Therapeutic drug monitoring (TDM) is essential, requiring rapid and accurate analytical methods for precise determination of drugs in biological material (typically plasma or serum). This study presents a novel capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) method designed for the simultaneous quantification of five penicillin antibiotics, two cephalosporins, one carbapenem, and two β-lactamase inhibitors in a single run. The method involves a simple sample pretreatment-precipitation with organic solvent-and has a run time of 20 min. Optimization of CZE separation conditions revealed that 20 mM ammonium hydrogen carbonate (NHHCO) serves as the optimal background electrolyte (BGE). Positive electrospray ionization (ESI) mode, with isopropyl alcohol (IP)/10 mM ammonium formate water solution (50/50, /) as the sheath liquid, was identified as the optimal condition for MS detection. Method validation according to the Food and Drug Administration (FDA) guideline for development of bioanalytical methods demonstrated satisfactory selectivity, linearity, recovery, robustness, and stability. The method's practicality was evaluated using the Blue Applicability Grade Index (BAGI), yielding a score of 77.5. Moreover, the greenness of the proposed method was evaluated by two commonly used metric tools-Analytical GREEnness (AGREE) and Green Analytical Procedure Index (GAPI). The developed CZE-MS/MS method offers a practical and reliable approach for quantifying a broad spectrum of β-lactam antibiotics in plasma. Its ability to simultaneously quantify multiple analytes in a single run, coupled with a straightforward sample pretreatment, positions it as a valuable and prospective tool for TDM in critically ill patients.
监测β-内酰胺类抗生素的血浆浓度至关重要，尤其是在重症患者中，浓度变化可能导致治疗失败或不良事件。标准化抗菌方案可能对所有患者都无效，特别是在生理参数改变的特殊群体中。药代动力学/药效学（PK/PD）研究突出了这些抗生素的时间依赖性抗菌活性，强调了个性化给药的必要性。治疗药物监测（TDM）至关重要，需要快速准确的分析方法来精确测定生物材料（通常是血浆或血清）中的药物。本研究提出了一种新型毛细管区带电泳-串联质谱（CZE-MS/MS）方法，该方法设计用于在一次运行中同时定量五种青霉素类抗生素、两种头孢菌素、一种碳青霉烯类抗生素和两种β-内酰胺酶抑制剂。该方法包括简单的样品预处理——用有机溶剂沉淀——运行时间为20分钟。CZE分离条件的优化表明，20 mM碳酸氢铵（NHHCO）是最佳背景电解质（BGE）。正电喷雾电离（ESI）模式，以异丙醇（IP）/10 mM甲酸铵水溶液（50/50，/）作为鞘液，被确定为MS检测的最佳条件。根据美国食品药品监督管理局（FDA）生物分析方法开发指南进行的方法验证显示出令人满意的选择性、线性、回收率、稳健性和稳定性。使用蓝色适用性等级指数（BAGI）评估了该方法的实用性，得分为77.5。此外，通过两种常用的度量工具——分析绿色度（AGREE）和绿色分析程序指数（GAPI）评估了所提出方法的绿色度。所开发的CZE-MS/MS方法为定量血浆中广谱β-内酰胺类抗生素提供了一种实用且可靠的方法。它能够在一次运行中同时定量多种分析物，再加上简单的样品预处理，使其成为重症患者TDM的一种有价值且有前景的工具。
