背景:这项研究的目的是开发和验证一种UPLC-MS/MS方法,用于同时测定13种AED,包括卡马西平,奥卡西平,拉莫三嗪,左乙拉西坦,托吡酯,普米酮,唑尼沙胺,加巴喷丁,拉科沙胺,Perampanel,普瑞巴林,鲁非酰胺,和vigabatrin,在全血样本中。
方法:建立了同时测定全血中13种AEDs的UPLC-MS/MS方法,并进行了准确性验证,精度,定量限(LOQ),基体效应,和稳定性。使用UPLC-MS/MS将我们的方法与两家不同的医院进行了比较。
结果:所有AED均在AMR(分析测量范围)范围内表现出线性,R2值范围从0.994到1.000。低和高质量控制(QC)水平的不精确性和不准确性在可接受的范围内,变异系数(CV)<15%。卡马西平的LOQ为0.62微克/毫升,奥卡西平为1.61微克/毫升,拉莫三嗪为1.30µg/mL,左乙拉西坦为13.20微克/毫升,topira-mate为1.26µg/mL,扑米酮1.01微克/毫升,唑尼沙胺为1.59微克/毫升,1.09µg/mL的拉科沙胺,加巴喷丁1.61微克/毫升,普瑞巴林为0.50微克/毫升,perampanel为0.07ng/mL,3.00µg/mL的鲁非酰胺,和2.06µg/mL的vigabatrin。所有AED都证明了可接受的残留测定参数,稳定性,和矩阵效应。此外,与两家不同的医院相比,该检测结果令人满意,偏倚小于15%。
结论:我们成功开发并验证了一种快速,稳健的UPLC-MS/MS方法,用于同时监测13种抗癫痫药物的常规治疗药物。
BACKGROUND: The goal of this study was to develop and validate a UPLC-MS/MS method for simultaneous mea-surement of 13 AEDs, including carbamazepine, oxcarbazepine, lamotrigine, levetiracetam, topiramate, primidone, zonisamide, gabapentin, lacosamide, perampanel, pregabalin, rufinamide, and vigabatrin, in whole blood samples.
METHODS: A UPLC-MS/MS method for simultaneous determination of 13 AEDs in whole blood was developed, and validation was conducted for accuracy, precision, limit of quantification (LOQ), matrix effect, and stability. Our method was compared to two different hospitals using UPLC-MS/MS.
RESULTS: All AEDs exhibited linearity across the AMR (analytical measurement range), with R2 values ranging from 0.994 to 1.000. The imprecision and inaccuracy for low and high quality control (QC) levels were within an acceptable range, with the coefficient of variation (CV) < 15%. The LOQ was 0.62 µg/mL for carbamazepine, 1.61 µg/mL for oxcarbazepine, 1.30 µg/mL for lamotrigine, 13.20 µg/mL for levetiracetam, 1.26 µg/mL for topira-mate, 1.01 µg/mL for primidone, 1.59 µg/mL for zonisamide, 1.09 µg/mL for lacosamide, 1.61 µg/mL for gabapentin, 0.50 µg/mL for pregabalin, 0.07 ng/mL for perampanel, 3.00 µg/mL for rufinamide, and 2.06 µg/mL for vigabatrin. All AEDs demonstrated acceptable assay parameters for carryover, stability, and matrix effects. Moreover, the assay showed satisfactory results compared to two different hospitals with a bias of less than 15%.
CONCLUSIONS: We successfully developed and validated a fast and robust UPLC-MS/MS method for routine therapeutic drug monitoring of thirteen antiepileptic drugs simultaneously.