采用毛细管电泳-激光诱导荧光检测法测定人血浆中的氨己烯酸。

Determination of vigabatrin in human plasma by means of CE with LIF detection.

作者信息

Musenga Alessandro, Mandrioli Roberto, Comin Irene, Kenndler Ernst, Raggi Maria Augusta

机构信息

Pharmaco-Toxicological Analysis Laboratory, Department of Pharmaceutical Sciences, Alma Mater Studiorum-University of Bologna, Via Belmeloro 6, I-40126 Bologna, Italy.

出版信息

Electrophoresis. 2007 Oct;28(19):3535-41. doi: 10.1002/elps.200700139.

DOI:10.1002/elps.200700139

PMID:17828800

Abstract

A method has been developed for the quantitation of the antiepileptic drug vigabatrin (VGB) in human plasma. It is based on CE with LIF detection. The effect of the pH of the buffer and of N-methylglucamine (GLC) as BGE constituent was investigated. The final BGE consisted of 50 mM borate buffer, pH 9.0, with 100 mM GLC and enabled separation within 12 min at 20 kV voltage. An SPE procedure was used for the pretreatment of biological samples, based on mixed-mode lipophilic-cation exchange cartridges, followed by a derivatization step with 6-carboxyfluorescein-N-succinimidyl ester (CFSE). Fluorescence was excited by an Ar-ion laser (lambda(exc) = 488 nm). Linearity was observed in the 10-120 microg/mL plasma concentration range. Extraction yield was >96%, precision (expressed as RSD) <6.7% and accuracy (recovery) was between 97.0 and 101.6%. The method has been successfully applied to the analysis of VGB in plasma of epileptic patients undergoing therapy with the drug.

摘要

已开发出一种定量测定人血浆中抗癫痫药物氨己烯酸（VGB）的方法。该方法基于毛细管电泳-激光诱导荧光检测（CE-LIF）。研究了缓冲液pH值以及作为背景电解质（BGE）成分的N-甲基葡糖胺（GLC）的影响。最终的BGE由pH 9.0的50 mM硼酸盐缓冲液与100 mM GLC组成，在20 kV电压下12分钟内实现分离。采用基于混合模式亲脂性阳离子交换柱的固相萃取（SPE）程序对生物样品进行预处理，随后用6-羧基荧光素-N-琥珀酰亚胺酯（CFSE）进行衍生化步骤。通过氩离子激光（激发波长λ(exc)=488 nm）激发荧光。在10 - 120 μg/mL血浆浓度范围内观察到线性关系。提取率>96%，精密度（以相对标准偏差RSD表示）<6.7%，准确度（回收率）在97.0%至101.6%之间。该方法已成功应用于接受该药物治疗的癫痫患者血浆中VGB的分析。