Rapid separation of peptides and proteins on 2-mu m porous microspherical reversed-phase silica material

The fast LC of peptides and proteins on a reversed-phase column packed with 2-mu m porous silica microspherical silica gel (TSKgel Super-ODS, 50 mm X 4.6 mm I.D.) was investigated. This short column packed with small particles showed high resolutions for peptides and proteins with high yields (over 80%). The relationship between the sample amount and peak area of a basic peptide indicated good linearity over a wide range of sample amount applied (1-500 ng). In comparison of the chromatograms of a tryptic digest obtained on porous and non-porous columns, many sharp peaks were recognized on the Super-ODS column owing to its high hydrophobicity. Using a short gradient time and high flow-rate, a peptide mixture could be separated successfully within 1 min.