Abstract: The method of Qtrap-ultra high performance liquid chromatography tandem mass spectrometry (Qtrap-UPLC-MS/MS) was established for simultaneous determination four amino acid neurotransmitters in rat’s brain tissue. The effects of powder, water decoction and dregs of Chloriti Lapis on amino acid content in PTZ-kindled epileptic rat’s brain tissue were studied. The contents of Glu, Gly, Asp and GABA in rat’s brain tissue (cortex and hippocampus) were detected by multiple reactions monitoring (MRM) scan mode in positive electrospray ionization (ESI⁺). The result shows that the four amino acids have good linear relationship, the RSDs of precision and accuracy conform to the requirements of analysis in biological samples. After rats were treated with different samples of Chloriti Lapis, excitatory neurotransmitter Glu, Asp in cerebral cortex and hippocampus decreased significantly. Compared with the model group, the content of Gly in cerebral cortex is increased in the percentage of total amino acidcontents, while the ratio of Asp/Gly and Glu/Gly are decreased. The contents of Glu, Asp, Gly and GABA in other groups display reduction in hippocampus compared with the model group. This shows that mineral Chinese medicine Chloriti Lapis can effectively reduce the excitatory amino acid contents in PTZ-kindled rat’s brain, and effectively regulate the content of Gly in the cortex and hippocampus.