Cobo-Golpe María, Paniagua-González Lucía, Lendoiro Elena, Blanco-Ces Miriam, López-Rabuñal Ángela, Abella Javier, Castro Dolores, Melcón Cristina, Fuentes Patricia, Carballeira Iria, García Carlos, Gómez Carmen, Lamas Manuel López-Rivadulla, Cruz Angelines, de-Castro-Ríos Ana
Toxicology Service, Institute of Forensic Sciences, C/San Francisco, s/n, 15782 Santiago de Compostela, Spain.
Neurology Service, University Hospital Complex of Ferrol, Av. da Residencia, 15405 Ferrol, Spain.
J Anal Toxicol. 2025 Jul 16. doi: 10.1093/jat/bkaf073.
Therapeutic drug monitoring (TDM) of antiepileptic drugs (AEDs) is used for optimization and individualization of patients' treatment. Capillary microsampling techniques are a promising alternative to conventional venous sampling for TDM. Both dried blood spots (DBS) and volumetric adsorptive microsampling (VAMS) devices are less invasive and patient-friendly sampling techniques which have been gaining interest in the last years. This study describes the development and validation of an LC-MS/MS method for the determination of 8 AEDs (Carbamazepine, Lacosamide, Levetiracetam, Lamotrigine, Phenobarbital, Valproic acid) and 2 metabolites (10,11-Dihydro-10-hydroxy-carbamazepine (DHCB) and carbamazepine-10,11-epoxide) in DBS and VAMS samples. The method was fully validated for linearity, selectivity, accuracy, precision, carryover, matrix effect, recovery and stability (15 days at room temperature and 72 h in autosampler). Moreover, the volume effect, volcano effect, and the hematocrit (Hct) effect were also assessed for DBS samples. All parameters showed satisfactory results, with a limit of quantification ranging from 0,5 to 10 µg/mL, depending on the analyte. Some instability issues were detected in DBS samples for oxcarbazepine. However, the inclusion of oxcarbazepine's metabolite DHCB overcomes this problem as it was stable under both conditions tested. Moreover, this is the first DBS or VAMS method reporting the inclusion of DHCB, which seems essential for the TDM of oxcarbazepine. The method was applied to 80 paired samples from patients under treatment with these drugs in order to study the suitability of the method for the detection of these compounds, and compare concentrations in paired VAMS, DBS, whole blood and plasma samples. Ratios between paired samples show a promising correlation between microsampling techniques and plasma concentrations.
抗癫痫药物（AEDs）的治疗药物监测（TDM）用于优化患者治疗并实现个体化。对于TDM而言，毛细管微量采样技术是传统静脉采样的一种有前景的替代方法。干血斑（DBS）和体积吸附微量采样（VAMS）装置都是侵入性较小且对患者友好的采样技术，近年来受到了关注。本研究描述了一种用于测定DBS和VAMS样品中8种抗癫痫药物（卡马西平、拉科酰胺、左乙拉西坦、拉莫三嗪、苯巴比妥、丙戊酸）和2种代谢物（10,11-二氢-10-羟基卡马西平（DHCB）和卡马西平-10,11-环氧化物）的液相色谱-串联质谱（LC-MS/MS）方法的开发与验证。该方法在线性、选择性、准确性、精密度、残留、基质效应、回收率和稳定性（室温下15天以及自动进样器中72小时）方面进行了全面验证。此外，还评估了DBS样品的体积效应、火山效应和血细胞比容（Hct）效应。所有参数均显示出令人满意的结果，定量限根据分析物的不同在0.5至10 μg/mL范围内。在DBS样品中检测到奥卡西平存在一些稳定性问题。然而，纳入奥卡西平的代谢物DHCB克服了这一问题，因为它在两种测试条件下均稳定。此外，这是首个报道纳入DHCB的DBS或VAMS方法，这对于奥卡西平的TDM似乎至关重要。该方法应用于80对正在接受这些药物治疗的患者的样品，以研究该方法检测这些化合物的适用性，并比较配对的VAMS、DBS、全血和血浆样品中的浓度。配对样品之间的比率显示出微量采样技术与血浆浓度之间有前景的相关性。
