The Influence of Retention on the Plate Height in Ion‐Exchange Chromatography

Abstract

The plate heights for the amino acid tyrosine (anion exchange) and the polypeptide aprotinin (cation exchange) were determined on a porous media (Resource 15) and a gel filled media (HyperD 20) at salt concentrations ranging from weak to strong retention. At a constant velocity, measurements showed that the plate height increase with increasing retention, went through a maximum, and finally, decreased as the retention increased, i.e., when the salt concentration was lowered further. The band broadening of a chromatographic peak in the column was caused by the axial dispersion and mass transfer. In this article, the rate of mass transfer in the particles is described by three different rate mechanisms, pore diffusion, solid diffusion, and parallel diffusion. The van Deemter equation was used to model the data to determine the mass-transfer properties. The development of the plate height with increasing retention revealed a characteristic behavior for each rate mechanism. In the pore diffusion model, the plate height increased toward a constant value at strong retention, while the plate height in the solid diffusion model decreased, approaching a constant value at strong retention. In the parallel diffusion model, both pore and solid diffusion took place. Therefore, the parallel diffusion model coincides with the pore diffusion model at weak retention and with the solid diffusion model at strong retention, while a maximum is reached at intermediate retention, resulting in a bell-shaped curve. This behavior corresponds to the observed variation of the plate height at constant velocity. Neither the pore nor the solid diffusion model can describe the experimental data while a satisfactory fit was obtained using the parallel diffusion model.

Design and control of integrated chromatography column sequences

“…Assuming that the separation in each chromatography step is well defined and each phase, its liquid volume and flow rate, are specified in column volumes, it is possible to scale the separation by assuming that the separation is unchanged if the residence time is constant . The amount eluted from the previous step has to be loaded on to the next step.…”

“…13 Assuming that the separation in each chromatography step is well defined and each phase, its liquid volume and flow rate, are specified in column volumes, it is possible to scale the separation by assuming that the separation is unchanged if the residence time is constant. 14,15 The amount eluted from the previous step has to be loaded on to the next step. For an adsorption column, the capacity has to be larger than the amount eluted from the previous column, constraint ii above, and for a gel filtration step, the load volume of the step has to be larger than the eluted volume, constraint i.…”