The invention belongs to the technical field of separation of chiral enantiomers, and in particular relates to a method for separating chiral enantiomers of Tiagabine hydrochloride by reversed-phase high performance liquid chromatography; a sulfobutyl-beta-cyclodextrin aqueous solution as an additive is added to an original mobile phase to obtain a desired reverse phase chromatographic mobile phase; the original mobile phase is acetonitrile, and the concentration of the sulfobutyl-beta-cyclodextrin aqueous solution is 0.04 g/ml, the volume ratio of the original mobile phase to the sulfobutyl-beta-cyclodextrin aqueous solution is 35:65, and the flow rate of the mobile phase is 1.0 ml/ml. The method can effectively avoid the defects that traditional beta-cyclodextrin is easy to cause blockage of a chromatographic column, and has the advantages of low cost, more economical application, wide measurement range and wide application prospect.