Derivatization chemistries for determination of serotonin, norepinephrine and dopamine in brain microdialysis samples by liquid chromatography with fluorescence detection
Derivatization chemistries for determination of serotonin, norepinephrine and dopamine in brain microdialysis samples by liquid chromatography with fluorescence detection
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DOI：
10.1002/bmc.560
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发表时间：
2006-03-01
影响因子：
1.8
通讯作者：
Yamaguchi, M
中科院分区：
文献类型：
--
作者：
Yoshitake, T;Kehr, J;Yamaguchi, M
本文综述了微透析样品中单胺类神经递质5-羟色胺（5-HT）、去甲肾上腺素（NE）和多巴胺（DA）的衍生化化学和微孔液相色谱荧光检测技术的研究进展。在温和的碱性条件下，5-羟基吲哚和儿茶酚胺与苄基胺（BA）反应，分别生成高荧光的2-苯基-4,5-吡咯啉苯并恶唑和2-苯基（4,5-二氢吡咯）[2,3-f]苯并恶唑。然而，对于DA的衍生化反应，与1,2-二苯乙二胺（DPE）的反应比与BA的反应具有更高的荧光强度，因此，为了同时测定脑微透析液中的5-HT， NE和DA，开发了与BA和DPE的两步衍生化反应。5-羟色胺、NE和DA在20 μ L注射量下的检出限分别为0.2、0.08和0.13 fmol，对应衍生化前标准溶液的浓度分别为30、12和19.5 pm。实验数据表明，该技术能够同时监测大鼠和小鼠大脑在基础条件下、药物治疗或生理刺激后的单胺类神经递质的神经释放池。这些技术在抑郁症、精神分裂症和帕金森病等精神和神经疾病的药物发现和临床研究中发挥着重要作用。版权所有(c) 2005 John Wiley & Sons, Ltd。
The present paper provides an overview on currently developed derivatization chemistries and techniques for determination of monoamine neurotransmitters serotonin (5-HT), norepinephrine (NE) and dopamine (DA) in microdialysis samples by microbore liquid chromatography with fluorescence detection. In mild alkaline conditions, 5-hydroxyindoles and catecholamines react with benzylamine (BA), forming highly fluorescent 2-phenyl-4,5-pyrrolobenzoxazoles and 2-phenyl(4,5-dihydropyrrolo) [2,3-f]benzoxazoles, respectively. However, for derivatization of DA a higher fluorescence intensity was achieved for reaction with 1,2-diphenylethylenediamine (DPE) rather than with BA, therefore for simultaneous determination of 5-HT, NE and DA in brain microdialysates, a two-step derivatization with BA followed by DPE was developed. The detection limits for 5-HT, NE and DA were 0.2, 0.08 and 0.13 fmol, respectively, in an injection volume of 20 mu L, which corresponds to concentrations of 30, 12 and 19.5 pm, respectively in standard solution prior to derivatization. The experimental data presented demonstrate the ability of the technique to simultaneously monitor neuronally releasable pools of monoamine neurotransmitters in the rat and mouse brains at basal conditions and following pharmacological treatments or physiological stimuli. These techniques play an important role in drug discovery and clinical investigation of psychiatric and neurological diseases such as depression, schizophrenia and Parkinson's disease. Copyright (c) 2005 John Wiley & Sons, Ltd.