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首页> 外文期刊>Journal of Neuroscience Methods >A high performance liquid chromatography method with electrochemical detection of gamma-aminobutyric acid, glutamate and glutamine in rat brain homogenates.
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A high performance liquid chromatography method with electrochemical detection of gamma-aminobutyric acid, glutamate and glutamine in rat brain homogenates.

机译：高效液相色谱法，电化学检测大鼠脑匀浆中的γ-氨基丁酸，谷氨酸和谷氨酰胺。

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摘要

Determination of gamma-aminobutyric acid (GABA), glutamate (Glu) and glutamine (Gln) in animal models has been important to understand the normal function and clinical aspects of some neurological diseases. Quantification of these amino acid transmitters has conventionally been performed by using a high performance liquid chromatography (HPLC) system. This paper describes an improved HPLC method with electrochemical detection for glutamate, glutamine and GABA determination in brain homogenates. The protocol is based on a precolumn derivatization of amino acids with o-phthalaldehyde and sodium sulfite, a separation through a C18, 5 microm particle size column and an isocratic elution. Several modifications of previous works on methanol percentage, pH, temperature, flow rate and derivatization solution concentration were done to obtain a suitable protocol for amino acid quantification in brain homogenate samples. Total elution time is 35 min approximately. Technical requirements and laboratory expenses of this new protocol are minimal. This technique showed high linearity, repeatability and accuracy.
机译：动物模型中γ-氨基丁酸（GABA），谷氨酸（Glu）和谷氨酰胺（Gln）的测定对于理解某些神经系统疾病的正常功能和临床方面具有重要意义。这些氨基酸递质的定量常规上已经通过使用高效液相色谱（HPLC）系统进行。本文介绍了一种改进的HPLC方法，可通过电化学方法检测脑匀浆中的谷氨酸，谷氨酰胺和GABA。该方案基于氨基酸与邻苯二甲醛和亚硫酸钠的柱前衍生化，通过C18、5微米粒径色谱柱的分离和等度洗脱。对甲醇含量，pH，温度，流速和衍生化溶液浓度进行了先前工作的几处修改，以获得用于脑匀浆样品中氨基酸定量的合适方案。总洗脱时间约为35分钟。此新协议的技术要求和实验室费用是最小的。该技术显示出高线性度，可重复性和准确性。

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