Evaluation of the Enantiomeric Separation of Dipeptides Using a Chiral Crown Ether LC Column

Abstract

Abstract The direct optical resolution of six dipeptides into four stereoisomers each was achieved on an enantioselective crown ether column. An inclusion complex is formed between the stationary phase and the solute when using an acidic mobile phase. The acidic mobile phase serves to protonate the requisite primary amine of the dipeptide thereby allowing an attractive interaction between the ammonium functional group and the oxygens of the crown ether. Due to the differences in stability of the complexes formed, the four optical isomers elute at different times allowing the stereoisomeric separation. One of the factors affecting enantioselectivity is the distance between the primary amine functional group and the stereogenic center of the chiral moiety. Dipeptides are particularly useful molecules for the studying this “distance effect” since the bonding order of the two amino acids can be reversed. In addition to the enantiomeric separations of dipeptides possessing two stereogenic centers, the behavior of dipeptide separations possessing only one chiral center (i.e., with achiral glycine as one of the residues) is examined to gain additional insight into the mechanism and the effect of the proximity of the primary amine group to the chiral center.