Laboratory of Clinical Neuropharmacology, Neurology Clinic, Department of Neurological Sciences, University of Bologna, Via Foscolo 7, 40123 Bologna, Italy.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Feb 1;878(3-4):461-5. doi: 10.1016/j.jchromb.2009.11.039. Epub 2009 Dec 3.
We present an implementation of a method we previously reported allowing the newer antiepileptic drugs (AEDs) rufinamide (RFN) and zonisamide (ZNS) to be simultaneously determined with lamotrigine (LTG), oxcarbazepine's (OXC) main active metabolite monohydroxycarbamazepine (MHD) and felbamate (FBM) in plasma of patients with epilepsy using high performance liquid chromatography (HPLC) with UV detection. Plasma samples (250 microL) were deproteinized by 1 mL acetonitrile spiked with citalopram as internal standard (I.S.). HPLC analysis was carried out on a Synergi 4 microm Hydro-RP, 250 mm x 4.6 mm I.D. column. The mobile phase was a mixture of potassium dihydrogen phosphate buffer (50 mM, pH 4.5), acetonitrile and methanol (65:26.2:8.8, v/v/v) at an isocratic flow rate of 0.8 mL/min. The UV detector was set at 210 nm. The chromatographic run lasted 19 min. Commonly coprescribed AEDs did not interfere with the assay. Calibration curves were linear for both AEDs over a range of 2-40 microg/mL for RFN and 2-80 microg/mL for ZNS. The limit of quantitation was 2 microg/mL for both analytes and the absolute recovery ranged from 97% to 103% for RFN, ZNS and the I.S. Intra- and interassay precision and accuracy were lower than 10% at all tested concentrations. The present study describes the first simple and validated method for RFN determination in plasma of patients with epilepsy. By grouping different new AEDs in the same assay the method can be advantageous for therapeutic drug monitoring (TDM).
我们介绍了一种方法的实施，该方法是我们之前报道的，允许使用高效液相色谱法（HPLC）与紫外检测法，将新型抗癫痫药物（AED）鲁非酰胺（RFN）和唑尼沙胺（ZNS）与拉莫三嗪（LTG）、奥卡西平的主要活性代谢物单羟基卡马西平（MHD）和非氨酯（FBM）同时在癫痫患者的血浆中进行定量分析。血浆样品（250 μL）用 1 mL 乙腈沉淀，加入西酞普兰作为内标（I.S.）。HPLC 分析在 Synergi 4 微米 Hydro-RP，250mm×4.6mm ID 柱上进行。流动相为磷酸二氢钾缓冲液（50mM，pH4.5）、乙腈和甲醇（65：26.2：8.8，v/v/v）的混合物，等度流速为 0.8mL/min。紫外检测器设置在 210nm。色谱运行时间为 19 分钟。通常同时开处方的 AED 不会干扰测定。两种 AED 的校准曲线在 RFN 的 2-40μg/mL 和 ZNS 的 2-80μg/mL 范围内均呈线性。两种分析物的定量限均为 2μg/mL，RFN、ZNS 和 I.S.的绝对回收率在 97%-103%之间。日内和日间精密度和准确度在所有测试浓度下均低于 10%。本研究描述了一种用于癫痫患者血浆中 RFN 测定的简单、验证方法。通过将不同的新型 AED 分组到同一个测定中，该方法有利于治疗药物监测（TDM）。
