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通过高效液相色谱与金纳米粒子引发的化学发光联用测定小鼠脑微透析液中的单胺类神经递质及其代谢产物。

Determination of monoamine neurotransmitters and their metabolites in a mouse brain microdialysate by coupling high-performance liquid chromatography with gold nanoparticle-initiated chemiluminescence.

作者信息

Li Na, Guo Jizhao, Liu Bo, Yu Yuqi, Cui Hua, Mao Lanqun, Lin Yuqing

机构信息

Department of Chemistry, University of Science and Technology of China (USTC), JinZhai Road No: 96, 230026 Hefei, Anhui, PR China.

出版信息

Anal Chim Acta. 2009 Jul 10;645(1-2):48-55. doi: 10.1016/j.aca.2009.04.050. Epub 2009 May 7.

Abstract

Our previous work showed that gold nanoparticles could trigger chemiluminescence (CL) between luminol and AgNO3. In the present work, the effect of some biologically important reductive compounds, including monoamine neurotransmitters and their metabolites, reductive amino acids, ascorbic acid, uric acid, and glutathione, on the novel CL reaction were investigated for analytical purpose. It was found that all of them could inhibit the CL from the luminol-AgNO3-Au colloid system. Among them, monoamine neurotransmitters and their metabolites exhibited strong inhibition effect. Taking dopamine as a model compound, the CL mechanism was studied by measuring absorption spectra during the CL reaction and the reaction kinetics via stopped-flow technique. The CL inhibition mechanism is proposed to be due to that these tested compounds competed with luminol for AgNO3 to inhibit the formation of luminol radicals and to accelerate deposition of Ag atoms on surface of gold nanoparticles, leading to a decrease in CL intensity. Based on the inhibited CL, a novel method for simultaneous determination of monoamine neurotransmitters and their metabolites was developed by coupling high-performance liquid chromatography with this CL reaction. The new method was successfully applied to determine the compounds in a mouse brain microdialysate. Compared with the reported HPLC-CL methods, the proposed method is simple, fast, and could determine more analytes. Moreover, the limits of linear ranges for NE, E, and DA using the proposed method were one order of magnitude lower than the luminol system without gold nanoparticles.

摘要

我们之前的工作表明,金纳米颗粒可引发鲁米诺与硝酸银之间的化学发光(CL)。在本工作中,为了分析目的,研究了一些具有生物学重要性的还原性化合物,包括单胺神经递质及其代谢物、还原性氨基酸、抗坏血酸、尿酸和谷胱甘肽,对这种新型CL反应的影响。发现它们均可抑制鲁米诺 - 硝酸银 - 金胶体体系的CL。其中,单胺神经递质及其代谢物表现出较强的抑制作用。以多巴胺作为模型化合物,通过测量CL反应过程中的吸收光谱以及采用停流技术研究反应动力学,对CL机理进行了研究。提出CL抑制机理是由于这些受试化合物与鲁米诺竞争硝酸银,抑制鲁米诺自由基的形成,并加速银原子在金纳米颗粒表面的沉积,导致CL强度降低。基于抑制型CL,通过将高效液相色谱与该CL反应联用,开发了一种同时测定单胺神经递质及其代谢物的新方法。该新方法成功应用于测定小鼠脑微透析液中的化合物。与已报道的HPLC - CL方法相比,该方法简单、快速,且能测定更多分析物。此外,使用该方法测定去甲肾上腺素(NE)、肾上腺素(E)和多巴胺(DA)的线性范围下限比无金纳米颗粒的鲁米诺体系低一个数量级。

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