Continuous rotating annular chromatography with partial recycling of the effluent (CRAC-PRE) was used for the separation of a ternary mixture of amino acids. A mathematical model describing the separation process was constructed to find the optimized operating conditions. The effectiveness of the CRAC-PRE for the separation of a ternary mixture was experimentally and theoretically elucidated by comparison to conventional CRAC. A recycling loop which gives reasonable separation was successfully arranged on the basis of the numerical simulation result. A combination of CRAC-PRE with the step elution method was applied to the complete separation of a ternary mixture of amino acids.