THE EFFECTS OF PORE DIAMETER AND LIGAND CHAIN-LENGTH ON FAST LIQUID-CHROMATOGRAPHY OF PROTEINS AND PEPTIDES

Fast flow separations of proteins and peptides were successfully carri ed out on microbore columns containing reversed phase supports of thre e pore diameters. Increasing flowrates from 0.25 to 3 ml/min (1.2 mm/s - 14.4 mm/s) did not show any adverse effects on the separations when SynChropak RPP-1000 and RPP-4000, which have 1000 Angstrom and 4000 A ngstrom pore diameters, were used. At 3 ml/min, the 1000 Angstrom and 4000 Angstrom, pore diameters yielded narrower peaks for proteins than the 300 Angstrom support, whereas the latter exhibited better resolut ion for peptides. Analyses in less than five minutes were achieved. A series of ligand chains (C-4, C-8 and C-18) showed few differences in retention or resolution for either protein or peptide standards. Longt erm stability of the 300 Angstrom and 4000 Angstrom supports was in ex cess of 25,000 column volumes when run at 3 ml/min with 0.1% trifluoro acetic acid.