Separation of optical isomers of dansyl amino acids by a reversed phase liquid chromatography has been accomplished by adding a copper (II) chelate of N-benzyl-L-proline to the mobile phase. The pH, the eluent composition and the concentration of copper (II) chelate all affect the optical separations. The elution orders between D and L DNS-amino acids were consistant except dansyl phenylalanine showing that D forms of DNS-amino acids elute earlier than L forms. These behaviors are different from the results obtained by the use of copper (II) proline. The retention mechanism for the optical separation of the dansyl amino acids can be explained by the equilibrium of liqand exchange and by hydrophobic interaction.