Kocur Arkadiusz, Moczulski Mateusz, Czajkowska Agnieszka, Częczek Anna, Rubik Jacek
Department of Drug Chemistry, Pharmaceutical and Biomedical Analysis, Faculty of Pharmacy, Medical University of Warsaw, 02-097 Warsaw, Poland.
Therapeutic Drug Monitoring, Clinical Pharmacokinetics and Toxicology Laboratory Unit, Department of Clinical Biochemistry, The Children's Memorial Health Institute, 04-730 Warsaw, Poland.
Eur J Pharm Sci. 2025 Sep 1;212:107200. doi: 10.1016/j.ejps.2025.107200. Epub 2025 Jul 13.
(Val)Ganciclovir (VGCV) is utilised for the prevention and treatment of cytomegalovirus (CMV) infections in pediatric patients who have undergone renal transplants. Due to toxic side effects, therapeutic drug monitoring (TDM) is recommended, especially in vulnerable populations. Microsampling offers collecting small volumes of body fluids (<50 µL), making it ideal for TDM service. This study aimed to develop and clinically validate two microsampling techniques - Mitra™ and Capitainer®- employing volumetric absorptive microsampling (VAMS) and quantitative dried blood spot (qDBS) methods, respectively. Additionally, reference methods for analysing GCV in whole blood and plasma were validated. The LC-MS/MS multi-matrix method for quantifying GCV was validated using a calibration range of 0.01-25 mg/L in accordance with ICH M10 and Food and Drug Administration guidelines. The calculated accuracies and precisions for all tested matrices were within 15 %. No significant matrix and carryover effects were observed. Clinical validation demonstrated the efficacy of the examined microsampling devices in practice, highlighting the feasibility of home-based self-sampling. The developed methodologies were utilised to assess GCV levels in clinical samples from pediatric renal transplant recipients receiving VGCV treatment within a preemptive or prophylactic CMV protocol. The results were clinically evaluated through regression and bias estimation analyses, confirming a high level of correlation between GCV levels in the tested matrices. In conclusion, the newly developed, straightforward, and robust multi-matrix LC-MS/MS method has been effectively validated and integrated into clinical practice. For the first time in the literature, we demonstrated the analytical and clinical feasibility of tested microsampling devices for routine GCV concentration monitoring.
（缬氨）更昔洛韦（VGCV）用于预防和治疗接受肾移植的儿科患者的巨细胞病毒（CMV）感染。由于存在毒副作用，建议进行治疗药物监测（TDM），尤其是在弱势群体中。微量采样可收集少量体液（<50µL），使其成为TDM服务的理想选择。本研究旨在开发并临床验证两种微量采样技术——分别采用体积吸收微量采样（VAMS）的Mitra™和采用定量干血斑（qDBS）方法的Capitainer®。此外，还验证了全血和血浆中更昔洛韦分析的参考方法。根据国际人用药品注册技术协调会（ICH）M10和美国食品药品监督管理局指南，使用0.01-25mg/L的校准范围对定量更昔洛韦的液相色谱-串联质谱（LC-MS/MS）多基质方法进行了验证。所有测试基质的计算准确度和精密度均在15%以内。未观察到显著的基质和残留效应。临床验证证明了所检查的微量采样设备在实际应用中的有效性，突出了家庭自我采样的可行性。所开发的方法用于评估接受VGCV治疗的儿科肾移植受者在抢先或预防性CMV方案下临床样本中的更昔洛韦水平。通过回归和偏差估计分析对结果进行临床评估，证实了测试基质中更昔洛韦水平之间的高度相关性。总之，新开发的、简单且稳健的多基质LC-MS/MS方法已得到有效验证并整合到临床实践中。我们首次在文献中证明了测试的微量采样设备用于常规更昔洛韦浓度监测的分析和临床可行性。
