Peptide separation in normal-phase liquid chromatography. Study of selectivity and mobile phase effects on various...

An experimental procedure for peptide separation by normal-phase liquid chromatography (NPLC) was proposed in previous papers. In the present study, the chromatographic behavior of amino, cyano, amide, diol and silica columns, which have been used in non-aqueous NPLC, is investigated anew. The amino column was not appropriate for peptide separation because of poor recovery. The cyano column could not be used due to lack of retention. The amide, diol and silica columns were useful for peptide separation. The chromatograms on amide, diol and silica columns were a little different when the mobile phase composition was changed. The recovery of peptides was good: diol > amide > silica. Repeatability and reproducibility using amide, diol and silica columns was satisfactory.