A novel operational mode for continuous rotating annular chromatography for the complete separation of a binary mixture is proposed. In this mode, the effluents in which two solutes overlap are continuously recycled into the inlet of the packed bed, while the pure effluents are directly recovered. The recycling manners are determined on the basis of the results obtained in the preliminary experiments and numerical simulation without recycling. The model mixture of binary amino acids is completely separated by this process combined with the cation exchange method. The recovered concentrations of the amino acids are larger than those from the conventional method. The effects of some of the other operational conditions on the resolution are experimentally and theoretically investigated.