Molecular definition of the retention parameters of peptides separated by RP-HPLC.

M. I. Aguilar, K. L. Richards, A. J. Round, M. T. Hearn

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Abstract

Different physicochemical properties of the constituent amino acid side chains have been employed in order to investigate the molecular basis of the experimentally derived retention parameters, S and log k0, of peptides separated by reversed-phase high performance liquid chromatography (RP-HPLC). The results demonstrate that the S-value is strongly related to the total surface area of a peptide but correlates poorly with the corresponding predicted hydrophobicity based on the summated amino acid group retention coefficients. In contrast, the experimentally derived log k0 value correlates well with both the total surface area and the predicted affinity values derived from the incremental free energy of transfer calculated from the summated amino acid retention coefficients. Since the S and log k0 values are parameters useful to the interpretation of peptide and protein chromatographic behavior in RP-HPLC, the results of the present study document the physicochemical relationships that exist between these retention parameters and peptide structure.

Original languageEnglish
Pages (from-to)207-217
Number of pages11
JournalPeptide Research
Volume7
Issue number4
Publication statusPublished - 1994

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