Determination of Glutamate and γ-Aminobutyric Acid in Rat Brain by HPLC with Pre Column Derivation and Electrochemical Detection

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.