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使用毛细管电泳和天然激光诱导荧光检测法分析脑微透析液中的血清素。

Analysis of serotonin in brain microdialysates using capillary electrophoresis and native laser-induced fluorescence detection.

作者信息

Benturquia Nadia, Couderc François, Sauvinet Valérie, Orset Cyrille, Parrot Sandrine, Bayle Christophe, Renaud Bernard, Denoroy Luc

机构信息

Laboratoire de Neuropharmacologie et Neurochimie, INSERM U512, Institut Fédératif des Neurosciences de Lyon (IFR 19), Université Claude Bernard, F-69373 Lyon Cedex 08, France.

出版信息

Electrophoresis. 2005 Mar;26(6):1071-9. doi: 10.1002/elps.200410150.

Abstract

Serotonin or 5-hydroxytryptamine (5-HT) is a major neurotransmitter in the central nervous system. In this work, a method for analyzing 5-HT in brain microdialysis samples using a commercially available capillary electrophoresis (CE) system has been developed. A pH-mediated in-capillary preconcentration of samples was performed, and after separation by capillary zone electrophoresis, native fluorescence of 5-HT was detected by a 266 nm solid-state laser. The separation conditions for the analysis of 5-HT in standard solutions and microdialysates have been optimized, and this method has been validated on both pharmacological and analytical bases. Separation of 5-HT was performed using a 80 mmol/L citrate buffer, pH 2.5, containing 20 mmol/L hydroxypropyl-beta-cyclodextrin (HP-beta-CD) and +30 kV voltage. The detection limit was 2.5 x 10(-10) mol/L. This method allows the in vivo brain monitoring of 5-HT using a simple, accurate CE measurement in underivatized microdialysis samples.

摘要

血清素或5-羟色胺(5-HT)是中枢神经系统中的一种主要神经递质。在本研究中,已开发出一种使用市售毛细管电泳(CE)系统分析脑微透析样品中5-HT的方法。对样品进行了pH介导的毛细管内预浓缩,通过毛细管区带电泳分离后,用266nm固态激光检测5-HT的天然荧光。对标准溶液和微透析液中5-HT分析的分离条件进行了优化,该方法已在药理学和分析学基础上得到验证。使用含20mmol/L羟丙基-β-环糊精(HP-β-CD)的80mmol/L柠檬酸盐缓冲液(pH 2.5)和+30kV电压进行5-HT的分离。检测限为2.5×10(-10)mol/L。该方法可通过对未衍生化微透析样品进行简单、准确的CE测量实现对脑内5-HT的体内监测。

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