Monitoring of antiepileptic drugs in children with epilepsy require multiple visits at a clinic for blood collection. Dried blood spot sampling is an alternative way of collection, performed at home by self-collection and can save time and costs for patients and family members. The aim was to develop and validate an LC–MS/MS dried blood spot method for carbamazepine, lamotrigine, levetiracetam and valproic acid with the requirements of using standard equipment and material in a routine laboratory setting.

Whatman-903 filter paper was utilized, and discs were punched into a 96 well plate with an automated puncher and barcode reading. Extraction with methanol/water solution including internal standards on an orbital shaker was followed by a vacuum centrifuge step and reconstitution in mobile phase. Bioanalytical validation was performed according to guidelines from European Medicines Agency and additional dried blood spot specific validation.

Calibration curves of the four included drugs had R² values ≥0.994. Therapeutic relevant concentrations were well within measuring ranges. Within and −between run precision had %CV:s of 2.9–10.5%. Accuracy (%bias) was between −16.5% (lower limit of quantification) to +7.4%. Blood spots in a volume range of 15–50 μL with hematocrit in expected ranges for this patient group were within precision and accuracy limits. To test the method, concentrations from dried blood spot venous and capillary patient samples (n = 50) were compared with plasma concentrations. Good correlations for all four drugs with R² of >0.92 was shown.

In summary, a fast method for dried blood spots based on a 96 well format was developed for four commonly prescribed antiepileptic drugs. This validated method with traceability in sample preparation by bar code reading makes it suitable for the clinical laboratory.

监测癫痫患儿的抗癫痫药物需要在诊所多次就诊以采集血液。干血点采样是一种替代的采集方式，可以在家中自行采集，可以节省患者和家庭成员的时间和成本。目的是开发和验证用于卡马西平，拉莫三嗪，左乙拉西坦和丙戊酸的LC-MS / MS干血斑方法，并要求在常规实验室环境中使用标准设备和材料。

使用Whatman-903滤纸，并用自动打孔器和条形码读取器将圆盘打孔到96孔板中。在定轨振荡器上用甲醇/水溶液（包括内标）萃取，然后进行真空离心步骤并在流动相中进行重构。根据欧洲药品管理局的指南进行生物分析验证，并进行额外的干血斑特异性验证。

所含四种药物的校准曲线的R ²值≥0.994。治疗相关浓度均在测量范围内。在运行精度之内和之间，％CV：s为2.9-10.5％。准确度（偏差百分比）在-16.5％（定量下限）至+ 7.4％之间。该患者组的血斑范围在15–50μL范围内，血细胞比容在预期范围内，均在精确度和准确度范围内。为了测试该方法，将干血点静脉和毛细血管病人样品（n = 50）中的浓度与血浆浓度进行了比较。显示所有四种药物的R ² > 0.92均具有良好的相关性。

总之，针对四种常用的抗癫痫药开发了一种基于96孔格式的干血斑快速方法。这种经过验证的方法可通过条形码读取在样品制备中具有可追溯性，使其适用于临床实验室。