High peak capacity separations of peptides in reversed-phase gradient elution liquid chromatography on columns packed with porous shell particles

The performance of 5 and 15 cm long columns packed with shell particles (Halo, AMT) is compared in gradient elution separations of the tryptic digests of myoglobin and bovine serum albumin. The influences of the temperature and the mobile phase flow rate on the column efficiency for two peptides are discussed. The influences of this flow rate, of the temperature, and of the gradient slopes on the peak capacities are also considered. Peak capacities in excess of 400 were achieved in 6 h with the longer column. Peak capacities of 200 could be achieved in 30 min with the shorter column.