Comparison of cyclodextrin-dipeptide inclusion complexes in the absence and presence of urea by means of capillary electrophoresis, nuclear magnetic resonance and molecular modeling

The use of capillary electrophoresis (CE) modified with cyclodextrin (CD) for the separation of stereoisomers of peptides is well established. To increase the solubility of beta-CD, urea is often added to the buffer which may influence the complexation of a CD with a guest molecule. The aim of the present study was to investigate the influence of urea on the complexation between dipeptides and beta-CD using Ala-Phe and Ala-Tyr as model compounds. For this purpose three different analytical methods were employed: capillary electrophoresis (CE), H-1-NMR spectroscopy and molecular dynamics simulations (MD). Electropherograms of the peptide enantiomers were different in the presence and absence of urea. For example, at pH2.5 in the absence of urea the enantiomers of Ala-Tyr are not separated in contrast to the use of buffers containing urea. Applying complexation-induced chemical shift (CICS) in NMR spectroscopy band rotating frame Overhauser enhancement spectroscopy (ROESY) revealed differences in the complexation. of the peptide enantiomers by beta-CD in the absence and presence of urea suggesting the stabilization of the complex through the phenolic hydroxyl group of tyrosine. MD simulations for different complexes were carried out with consideration of both water and urea molecules in solution. Simulations were performed for 1 ns. In conclusion, NMR spectroscopy and MD methods help to understand the structure of peptide-CD complexes and the separation and migration behavior in CE. ((c) Wiley-VCH Verlag GmbH & Co.