Separation of dipeptide and tripeptide enantiomers in capillary electrophoresis using carboxymethyl-beta-cyclodextrin and succinyl-beta-cyclodextrin: influence of the amino acid sequence, nature of the cyclodextrin and pH.

Abstract

The separation of the LL and DD enantiomers of dipeptides and tripeptides using cyclodextrins (CDs) containing carboxyl groups was investigated with respect to the amino acid sequence of the peptides, the nature of the cyclodextrin and the buffer pH. Compared to succinyl-beta-cyclodextrin, carboxymethyl-beta-cyclodextrin was the more universal CD for enantioseparations. Reversal of the enantiomer migration order upon increasing the buffer pH from 2.5 to 3.5 was observed in some cases. As shown for Phe-Phe reversal of the migration order also occurred between pH 3.5 and 5.3. Complexation constants and complex mobilities change with pH as both, the charge of the peptide and the charge of the CD vary depending on the pH. The complexation constants and complex mobilities of the dipeptides Ala-Phe and Phe-Phe were determined in order to explain the enantiomer migration behavior in the pH range 2.5-5.3. While the complexation constants determined the migration order at pH 2.5 and 5.3, complex mobility had a strong influence around pH 3.5-3.8.