Abstract: OBJECTIVE To establish a determination method for 5 kinds of amino acid in rats with high performance liquid-fluorescence detector. To study the effects of tetramethylpyrazine phosphate(TMPP) on amino acid content in acute cerebral ischemia rats model. METHODS Microdialysis fluid was determined with Agilent 1100 liquid system fluorescence detector, ZORBAX SB-Aq C_l8 chromatographic column was applied. Excitation wavelength and emission wavelength were set at 357 nm and 455 nm, respectively, mobile phase consisted of methanol and 50 mmol·L^-1 sodium acetate buffer (pH=6.5), with gradient elution, the flow rate was 1 mL·min^-1. Tweleve male rats randomly divided into model control group and treatment group, each group had 6. Microdialysis was used to collect samples in striatum of rats area after dialysate, five kinds of amino acid concentration was determined by HPLC-FD. RESULTS Glu and Asp levels in TMPP group were decreased significantly compare with control group, however, the amount of Tau and GABA increased. CONCLUSION TMPP can obviously reduce excitatory amino acid content and increase brain inhibitory amino acid content in model rats, which has protect effect on acute ceretral ischemia rats.