Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany.
Institute of Laboratory Medicine, University Hospital, LMU Munich, Germany.
J Pharm Biomed Anal. 2019 Mar 20;166:398-405. doi: 10.1016/j.jpba.2019.01.038. Epub 2019 Jan 25.
The aim of this project was to develop and validate an isotope-dilution liquid chromatography high resolution mass spectrometry (LC-HRMS) method for the quantification of the 11 most widely used systemic antimycotics and to study whether HRMS is a feasible alternative for therapeutic drug monitoring (TDM) when compared to tandem MS (MS/MS) technology.
After protein precipitation, followed by automated online sample clean-up the analytes were separated within 4 min on a C18 column using an acetonitrile-water gradient. Eleven antimycotics, namely 5-flucytosine, amphotericin B, anidulafungin, fluconazole, isavuconazole, itraconazole, ketoconazole, micafungin, OH-itraconazole, posaconazole and voriconazole were finally quantified in full MS scan mode using positive electrospray ionization (ESI +) with a mass range fromm/z 110-1300 using HRMS. The method was comprehensively validated on the basis of the European Medicines Agengy (EMA) method validation protocol using commercially available IVD kit components.
Good linear relationship between peak area responses and drug concentrations (R > 0.995) and excellent selectivity were observed for all antimycotics in this study. Inaccuracy and imprecision of all quality controls were consistently below ± 12.6% and ± 8.1%, respectively. Quantification results were in agreement with an IVD LC-MS/MS method.
HRMS was shown to be suitable for TDM of small molecules when compared to tandem mass spectrometry. The novel HRMS method is quickly installed and may be a robust and reliable tool for routine TDM of antimycotics in clinical laboratories.
本项目的目的是开发和验证一种同位素稀释液相色谱高分辨质谱（LC-HRMS）方法，用于定量分析 11 种最广泛使用的全身抗真菌药物，并研究 HRMS 是否可以替代串联质谱（MS/MS）技术进行治疗药物监测（TDM）。
在蛋白质沉淀后，通过自动在线样品净化，在 C18 柱上使用乙腈-水梯度在 4 分钟内分离分析物。使用正电喷雾电离（ESI +）在全 MS 扫描模式下，最终在正离子模式下定量 11 种抗真菌药物，质量范围为 m/z 110-1300，使用 HRMS。该方法是根据欧洲药品管理局（EMA）方法验证方案，使用商业上可获得的 IVD 试剂盒组件进行全面验证的。
在本研究中，所有抗真菌药物的峰面积响应与药物浓度之间均观察到良好的线性关系（R>0.995）和出色的选择性。所有质控品的准确度和精密度均始终保持在±12.6%和±8.1%以下。定量结果与 IVD LC-MS/MS 方法一致。
与串联质谱相比，HRMS 适合于小分子的 TDM。与传统的 MS/MS 方法相比，该新型 HRMS 方法具有快速安装的优势，可能成为临床实验室常规 TDM 抗真菌药物的一种强大且可靠的工具。
