Abstract: The separation processing of trehalose and glucose by simulated moving bed chromatography was studied.Related parameters were measured in single column experiment firstly.And the complete separation zone was found with triangle theory under linear condition.Higher purity of the trehalose (97.6%) and glucose (98.8%) were obtained through optimizing in SMB actual operation.Optimal SMB parameters were switching time 11min, feeding rate 13.8mL/min, desorbent solution flow rate 29.7mL/min, extraction solution flow rate 20mL/min, raffinate flow rate 23.5mL/min, and cycling flow rate 17.6mL/min.System operating temperature was 58℃.