This work concentrates on the separation mechanism and application of chiral ligand exchange high-speed countercurrent chromatography in enantioseparation of ten racemic aromatic α-hydroxyl acids, including mandelic acid, 2-chloromandelic acid, 4-methoxymandelic acid, 4-hydroxymandelic acid, α-methylmandelic acid, 4-hydroxy-3-methoxy-mandelic acid, 3-chloromandelic acid, 4-bromomandelic acid, α-cyclopentylmandelic acid and α-cyclohexylmandelic acid, in which five of the racemates were successfully enantioseparated by analytical apparatus with an optimized solvent system. The two-phase solvent system was composed of butanol-water (1:1, v/v) or hexane-n-butanol-water (0.5:0.5:1, v/v), to which N-n-dodecyl-l-proline was added in the organic phase as chiral ligand and cupric acetate was added in the aqueous phase as a transition metal ion. Various influence factors in high-speed countercurrent chromatography were optimized by enantioselective liquid-liquid extraction. The separation mechanism for chiral ligand exchange high-speed countercurrent chromatography was proposed based on the results of present studies. Successful enantioseparations of 72mg of mandelic acid, 76mg of 2-chloromandelic acid and 74mg of 4-methoxymandelic acid were achieved individually with high resolution by preparative high-speed countercurrent chromatography. The HPLC purity of all enantiomers was over 96% with the recovery in the range of 82-90% from the collected fractions.