Focusing Review

Study of HILIC retention behavior using Hydrophilic retention coefficients

Tatsunari Yoshida

Analytical Applications Department, Shimadzu Corporation, 380-1, Horiyamashita, Hadano-city, Kanagawa, 259-1304, Japan

Abstract:

Hydrophilic interaction chromatography (HILIC) is able to retain and separate hydrophilic peptides that are not retained on an octadecyl silica column in reversed.phase liquid chromatography (RPLC). It has been reported that separation selectivities in HILIC and RPLC methods differed significantly. In order to clarify the differences of selectivities, a set of “hydrophilicity retention coefficients” in HILIC of peptide is presented. The contribution of each amino acid residue to retention on HILIC column was calculated by linear multiple regression analysis using 121peptide retention times. A set of the hydrophilicity retention coefficients on each column successfully explained the contribution degree of retention. In this review, recent developments in the separation of peptides by HILIC using polar sorbents and less polar eluents, and the discussion of retention selectivities using hydrophilicity retention coefficients, have been summarized.

Keywords: hydrophilicity retention coefficients, selectivity, hydrophilicity parameters, HILIC, peptide separation

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