Department of Drug Chemistry, Pharmaceutical and Biomedical Analysis, Medical University of Warsaw, Banacha 1, 02-097 Warsaw, Poland.
Therapeutic Drug Monitoring, Clinical Pharmacokinetics and Toxicology Laboratory, Department of Clinical Biochemistry, The Children's Memorial Health Institute in Warsaw, Dzieci Polskich 20, 04-730 Warsaw, Poland.
Int J Mol Sci. 2024 Aug 12;25(16):8760. doi: 10.3390/ijms25168760.
Ganciclovir (GCV) and its prodrug valganciclovir (VGCV) are antiviral medications primarily used to treat infections caused by cytomegalovirus (CMV), particularly in immunocompromised individuals such as solid organ transplant (SOT) recipients. Therapy with GCV is associated with significant side effects, including bone marrow suppression. Therefore, therapeutic drug monitoring (TDM) is mandatory for an appropriate balance between subtherapeutic and toxic drug levels. This study aimed to develop and validate three novel methods based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for GCV determination in serum (reference methodology), dried serum spots (DSS), and VAMS-Mitra™ devices. The methods were optimized and validated in the 0.1-25 mg/L calibration range. The obtained results fulfilled the EMA acceptance criteria for bioanalytical method validation. Assessment of DSS and VAMS techniques extended GCV stability to serum for up to a minimum of 49 days (at room temperature, with desiccant). Developed methods were effectively evaluated using 80 clinical serum samples from pediatric renal transplant recipients. Obtained samples were used for DSS, and dried serum VAMS samples were manually generated in the laboratory. The results of GCV determination using serum-, DSS- and VAMS-LC-MS/MS methods were compared using regression analysis and bias evaluation. The conducted statistical analysis confirmed the interchangeability between developed assays. The DSS and VAMS samples are more accessible and stable during storage, transport and shipment than classic serum samples.
更昔洛韦（GCV）及其前体药物缬更昔洛韦（VGCV）是主要用于治疗巨细胞病毒（CMV）感染的抗病毒药物，特别是在免疫功能低下的个体中，如实体器官移植（SOT）受者。GCV 治疗与显著的副作用相关，包括骨髓抑制。因此，治疗药物监测（TDM）对于在治疗和毒性药物水平之间取得适当平衡是强制性的。本研究旨在开发和验证三种基于液相色谱-串联质谱（LC-MS/MS）的新方法，用于血清（参考方法）、干血清斑（DSS）和 VAMS-Mitra™设备中 GCV 的测定。该方法在 0.1-25mg/L 的校准范围内进行了优化和验证。获得的结果满足 EMA 对生物分析方法验证的验收标准。DSS 和 VAMS 技术的评估将 GCV 稳定性扩展至血清中，至少可达 49 天（在室温下，使用干燥剂）。使用来自儿科肾移植受者的 80 份临床血清样本对开发的方法进行了有效评估。获得的样本用于 DSS，并且在实验室中手动生成干燥的血清 VAMS 样本。使用血清、DSS 和 VAMS-LC-MS/MS 方法测定 GCV 的结果通过回归分析和偏差评估进行比较。进行的统计分析证实了开发的测定方法之间的可互换性。与经典的血清样本相比，DSS 和 VAMS 样本在储存、运输和运输过程中更易于获得且更稳定。
