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目的 :建立大鼠脑组织内谷氨酸和γ 氨基丁酸的HPLC测定方法。方法 :大鼠脑组织匀浆液低温离心 ,上清液经甲醇沉淀蛋白并稀释至合适浓度后与邻苯二醛在亚硫酸钠存在下发生衍生化反应 ,衍生物经HPLC分离 ,电化学检测。结果 :谷氨酸在 0 .1～ 2 .0 μg·ml- 1 ,γ 氨基丁酸在 0 .0 5～ 1.0 μg·ml- 1 浓度范围内 ,线性关系良好。谷氨酸测定的日内和日间变异系数(RSD )分别小于 3.4%和 4.5 % ,γ 氨基丁酸测定的日内和日间变异系数 (RSD )分别小于 4.2 %和 8.6 %。两者回收率分别为95 .9%～ 10 2 .3%和 10 7.2 %～ 110 .0 %。谷氨酸和γ 氨基丁酸的最小检出浓度均为 5ng·ml- 1 。SD大鼠小脑、皮层、丘脑、海马内谷氨酸的含量分别为 96 0 .2、10 6 8.0、46 0 .2和 846 .6 μg·g- 1 ;γ 氨基丁酸的含量分别为 186 .0、16 5 .4、149.3和 188.6 μg·g- 1 。结论 :本法简便、灵敏、准确 ,可用于大鼠脑组织内谷氨酸和γ 氨基丁酸的测定
Objective: To establish a method for the determination of glutamate and γ-aminobutyric acid in rat brain. Methods: The homogenate of rat brain was centrifuged at low temperature. The supernatant was precipitated with methanol and diluted to the proper concentration. The derivatization reaction was carried out with o-phthalaldehyde in the presence of sodium sulfite. The derivatives were separated by HPLC and electrochemically detected. Results: The linearity of glutamic acid was in the range of 0.1-2.0 μg · ml-1 and γ-aminobutyric acid was in the range of 0.5-0.1 μg · ml-1. The intra-and inter-day coefficient of variation (RSD) for glutamic acid was less than 3.4% and 4.5%, respectively, and the intra-and inter-day coefficient of variation (CV) for gamma- aminobutyric acid was less than 4.2% and 8.6%, respectively. The recoveries of the two were 95.9% -102.3% and 10 7.2% -110.0%, respectively. The minimum detectable concentrations of glutamic acid and γ-aminobutyric acid were 5 ng · ml-1. The contents of glutamic acid in cerebellum, cortex, thalamus and hippocampus of SD rats were 96.02, 106.0, 426.0 and 846.6 μg · g-1, respectively, and the contents of γ-aminobutyric acid were 186 .0,16 5 .4,149.3 and 188.6 μg · g-1. Conclusion: This method is simple, sensitive and accurate and can be used for the determination of glutamate and γ-aminobutyric acid in rat brain