摘要
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
Aim To develop a HPLC method to determine glutamate and γ-aminobutyric acid in rat brain tissue.Methods Homogenate of rat brain tissue was centrifuged at low temperature.Supernatant was deproteinized and dilluted to a suitable concentration before reaction with OPA in the presence of sulfite ions.Derivatives were seperated with HPLC and detected with ECD.Results A good linerity was preseuted from 0.1 to 2.0μg·ml -1 for glutamate,as well as from 0.05 to 1.0μg·ml -1 for γ-aminobutyric acid.Within day and between day RSD of the method was less than 3.4% and 4.5% for glutamate,4.2% and 8.6% for γ-aminobutyric acid, respectively.The recoveries of glutamate and γ-aminobutyric acid was 95.9%～102.3% and 107.2%～110.0%.The content of glutamate in cerebellum,cerebral cortex,thalamus and hippocampus of normal SD rat was 960.2,1068.0,460.2,846.6μg·g -1 and the content of γ-aminobutyric acid was 186.0,165.4,149.3,188.6μg·g -1 ,respectively.Conclusion The method is simple,sensitive and accurate.It can be used in the determination of glutamate and γ-aminobutyric acid in rat brain tissue.
出处
《解放军药学学报》
CAS
2000年第6期299-302,共4页
Pharmaceutical Journal of Chinese People's Liberation Army
关键词
HPLC
柱前衍生化
电化学
谷氨酸
Γ-氨基丁酸
HPLC
Pre-column derivation
Electrochemical detection
Glutamate
γ-Aminobutyric a,
