Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Bad Oeynhausen, Germany.
Talanta. 2013 Jun 15;110:71-80. doi: 10.1016/j.talanta.2013.02.010. Epub 2013 Feb 13.
Therapeutic drug monitoring (TDM) may be very useful in the clinical management of antiepileptic drug therapy for multiple reasons, such as individual variability, metabolism, genetic factors or drug-drug or drug-food interactions. In addition, TDM is helpful to study the variation in pharmacokinetics that occurs between individuals. Here, we describe a rapid assay using ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry to measure the antiepileptic drugs lacosamide, lamotrigine, levetiracetam, primidone, topiramate, and zonisamide. After the addition of internal standards (ISs) and protein precipitation of serum or plasma, 1 μl of sample was separated on a 2.1×50 mm reverse phase column (Waters, Acquity UPLC BEH Phenyl, 1.7 μm). Analytes were then ionized and detected by electrospray ionization mass spectrometry with multiple reaction monitoring. Runtime was 2.5 min per injection. Matrix effects were investigated by systematical ion suppression and in-source fragmentation experiments. The calibration curves of the 6 antiepileptic drugs were linear over the working range between 0.05 and 50 mg/L (r>0.99). The limit of detection (LOD) was <0.05 mg/L, whereas the limit of quantification (LLOQ) was 0.10 mg/L of all drugs measured in the assay. The intraassay and interassay coefficients of variation for all compounds were <15% for very low concentration (0.1 mg/L) and <8% in the clinically relevant concentration range (>1.0 mg/L). Mean recoveries were between 87.8 and 98.6% for all drugs. There were no significant ion suppressions detected at the elution times of the analytes. The mean differences between serum and heparinized plasma values were less than 6% for the 6 antiepileptic drugs. All drugs were stable in serum at -20°C, 4°C, and even at RT for at least 1 month. In summary, a specific and sensitive stable isotope dilution UPLC-MS/MS method was developed and validated for routine clinical monitoring of lacosamide, lamotrigine, levetiracetam, primidone, topiramate, and zonisamide.
治疗药物监测（therapeutic drug monitoring，TDM）可能由于个体差异、代谢、遗传因素或药物-药物或药物-食物相互作用等多种原因，在抗癫痫药物治疗的临床管理中非常有用。此外，TDM 有助于研究个体之间发生的药代动力学变化。在这里，我们描述了一种使用超高效液相色谱-电喷雾串联质谱法测量抗癫痫药物拉科酰胺、拉莫三嗪、左乙拉西坦、扑米酮、托吡酯和唑尼沙胺的快速分析方法。在加入内标（IS）并沉淀血清或血浆中的蛋白质后，取 1μl 样品在 2.1×50mm 反相柱（Waters， Acquity UPLC BEH Phenyl，1.7μm）上进行分离。然后，通过电喷雾电离质谱法和多反应监测对分析物进行离子化和检测。每个进样的运行时间为 2.5 分钟。通过系统的离子抑制和源内碎裂实验研究了基质效应。6 种抗癫痫药物的校准曲线在 0.05 至 50mg/L 的工作范围内呈线性（r＞0.99）。所有药物的检测限（LOD）均＜0.05mg/L，定量下限（LLOQ）均为 0.10mg/L。所有化合物在非常低浓度（0.1mg/L）和临床相关浓度范围内（＞1.0mg/L）的日内和日间变异系数均＜15%。所有药物的平均回收率均在 87.8%至 98.6%之间。在分析物的洗脱时间没有检测到明显的离子抑制。6 种抗癫痫药物的血清和肝素化血浆值之间的平均差异小于 6%。所有药物在 -20°C、4°C 甚至室温下至少 1 个月均稳定。总之，开发并验证了一种用于拉科酰胺、拉莫三嗪、左乙拉西坦、扑米酮、托吡酯和唑尼沙胺常规临床监测的特异性和灵敏性稳定同位素稀释 UPLC-MS/MS 方法。
