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采用超快速液相色谱-电化学检测法分析脑单胺神经递质的快速灵敏方法。

A rapid and sensitive method for the analysis of brain monoamine neurotransmitters using ultra-fast liquid chromatography coupled to electrochemical detection.

机构信息

INSERM U1028, Lyon Neuroscience Research Center, Ani-RA NeuroChem, Lyon, France.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Dec 15;879(32):3871-8. doi: 10.1016/j.jchromb.2011.10.038. Epub 2011 Nov 3.

DOI:10.1016/j.jchromb.2011.10.038
PMID:22100552
Abstract

Electrochemical detection is often used to detect catecholamines and indolamines in brain samples that have been separated by conventional reverse-phase high performance liquid chromatography (HPLC). This paper presents the transfer of an existing chromatographic method for the determination of monoamines in brain tissues using 5 μm granulometry HPLC columns to columns with a particle diameter less than 3 μm. Several parameters (repeatability, linearity, accuracy, limit of detection, and stability of samples) for this new ultrafast high performance liquid chromatography (UHPLC) method were examined after optimization of the analytical conditions. The separation of seven compounds, noradrenaline, dopamine and three of its metabolites, dihydroxyphenylacetic acid, homovanillic acid, and 3-methoxytyramine, and serotonin and its metabolite, 5-hydroxyindole-3-acetic acid was analyzed using this UHPLC-electrochemical detection method. The final method, which was applied to brain tissue extracts from mice, rats, and cats, decreased analysis time by a factor of 4 compared to HPLC, while guaranteeing good analytical performance.

摘要

电化学检测常用于检测经传统反相高效液相色谱 (HPLC) 分离的脑样本中的儿茶酚胺和吲哚胺。本文介绍了将现有的用于测定脑组织中单胺类物质的色谱方法从使用 5μm 粒径 HPLC 柱转移到粒径小于 3μm 的柱上。在优化分析条件后,对这种新的超快速高效液相色谱 (UHPLC) 方法的几个参数(重复性、线性、准确性、检测限和样品稳定性)进行了检验。使用这种 UHPLC-电化学检测方法分析了七种化合物,包括去甲肾上腺素、多巴胺及其三种代谢物,二羟苯乙酸、高香草酸和 3-甲氧基酪胺,以及血清素及其代谢物 5-羟色胺-3-乙酸。最终方法应用于从小鼠、大鼠和猫的脑组织提取物中,与 HPLC 相比,分析时间缩短了 4 倍,同时保证了良好的分析性能。

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