Hydrodynamic Fractionation of Macromolecules. I. A Simple Theory

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Abstract
A new mechanism for separation in gel permeation chromatography (CPC) or gel filtration has been developed based upon the postulate that flow occurs through the gel phase. On the basis of the hydrodynamic nature of the separation mechanism, the new name Hydrodynamic Fractionation is proposed. A theory has been formulated for the case of equal pore size in the gel phase and equal sized spaces between the beads. This theory predicts the elution volume vs molecular weight curve. The flow-rate dependence of the separation peaks is investigated. The theoretically predicted relationship between molecular weight and elution volume compares very well with the gel permeation experiments using glass beads as packing probably because these packings correspond most closely to the assumptions in the theory. In the cross-linked polymer packing where a large distribution of pore sizes exists the theory does not fit well; however, it does predict the general shape of the curve. The flow-rate dependence of the separation peaks is experimentally the same as qualitatively predicted. Also the experimental findings of equilibrium experiments were correlated with the corresponding GPC data using the present theory.

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