Determination of the lipophilicity of some peptides. Effect of surface pH of silica.

The lipophilicities of 22 peptides were determined by reversed-phase thin-layer chromatography using methanol as organic modifier in the concentration range 0-90 vol.-% on silica supports with surface pH values of 2.0,4.5,6.0,7.5 and 9.0. Only one of the 22 peptides (Trp-Ala-Ile) followed the general rule, i.e., its RM value decreased linearly with increasing proportion of organic modifier over the whole concentration range. Most peptides exhibited typical silanophilic retention behaviour, the RM value decreasing with increasing organic phase concentration in the lower concentration range and then increasing with further increase in the proportion of organic modifier. In some instances the lipophilicity increased linearly with increasing proportion of methanol. The silanophilic effect depended not only on the structure of the peptide, but also on the surface pH of the silica support. The retention behaviour of peptides can be well described by a polynomial function, the linear and quadratic forms of methanol concentration and surface pH being the independent variables. Principal component analysis showed that the presence of a ring structure in the peptides has the greatest impact on their retention behaviour, the overall polarity (basic, neutral or acidic) being of secondary importance. The number of amino acids in the peptide has a negligible effect on the chromatographic behaviour.