Abstract: In order to improve the microorganism fermentation process of producing glutamine，an electrodialysis method was developed to separate glutamic acid and glutamine. The feasibility of this method was predicted through calculating cation and anion concentrations in typical amino acid mixture solutions，and then validated by electrodialysis experiment. Furthermore，the influence of ammonium sulfate on the separation of amino acids mixture was investigated. Through the theoretical prediction and experiment tests，electrodialysis method could be used directly to separate glutamic acid and glutamine，and a little ammonium sulfate in the mixture benefited the separation process.