Analysis of dispersion mechanism in gel chromatography. II. Dispersion mechanism in gel chromatography of proteins.

Abstract

The dispersion mechanism in gel chromatography of proteins was quantitatively studied on the basis of a dispersion model which gave consideration to the contribution of gel phase diffusion and longitudinal dispersion in a column. Particularly, the effect of gel phase diffusion was examined in detail by comparing the values determined by applying the moment analysis to the elution curve and those obtained for a single gel bead.When the distribution coefficient of solute was small and the particle size of gel was large, the elution curve was asymmetrical and had a prolonged tail of low concentration. The gel phase diffusion coefficient determined from the elution curve was much higher than that for a single gel bead owing to the effect of the tail. This suggested that only a small portion near the gel bead surface was accessible to most of the solute applied owing to the much lower diffusion velocity in gel than the superficial velocity. The column efficiency improved as compared with the theoretical result...