Abstract: Based on chiral ligand exchange, extraction and separation of α-cyclohexyl-mandelic acid (HCHMa) enantiomers in the two-phase system containing copper ion (II) (Cu2+) and N-n-dodecyl-L-hydroxyproline (HN) was studied. The influence of pH, concentrations of Cu2+ and chiral ligand and solvents on partition coefficients (K) and separation factors (α), were discussed in detail, respectively. The experimental results show that HN forms more stable ternary complexs with S-HCHMa enantiomer than with R-enantiomer. There is a larger influence of pH on K and α. At low pH values (<3.5), formation of binary complexes is thermodynamically unfavourable. K and α are best when pH values are above 3.5 and the molar ratio of chiral ligand to Cu2+ is 2:1. Solvents also influence K and α very much.