Abstract
Commercial ovalbumin was measured by high-performance ion-exchange chromatography on TSK-GEL IEX-545 DEAE SIL and linear gradient elution with NaCl to investigate the effects of gradient, flow-rate, column length and sample loading on resolution and retention. As the gradient became less steep, both resolution and separation time increased. At constant gradient time, the resolution increased and the separation time decreased with increasing flow-rate, but at constant gradient volume, both resolution and separation time increased with decreasing flow-rate. Columns of 75 mm and 150 mm in length provided almost identical resolution. The maximum sample loading was approximately 0.5 mg for columns of 6 mm I.D. and almost independent of column length and injection volume.
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