The invention belongs to the technical field of the sugar industry and discloses a method for separating mycose from glucose. According to the method, nano-gold is as a catalyst and the mixed liquid of mycose and glucose is as a substrate, wherein the nano-gold catalyzes glucose in the mixed liquid to generate gluconic acid; and then the gluconic acid is separated from the mycose through a chromatographic process. The glucose is catalyzed by the nano-gold to generate the gluconic acid, and the sugar-to-sugar separation applied to mycose and glucose is transformed into sugar-to-acid separation, so that the separation efficiency is greatly improved; besides, the method is simple and quick, wide in raw material source and low in cost; therefore, the method is an efficient new process for separating mycose from glucose.