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Biology LibreTexts

9.3: Ion Exchange Chromatography

In ion exchange chromatography, the support consists of tiny beads to which are attached chemicals possessing a charge. Each charged molecule has a counter-ion. The figure shows the beads (blue) with negatively charged groups (red) attached. In this example, the counter-ion is sodium, which is positively charged. The negatively charged groups are unable to leave the beads, due to their covalent attachment, but the counter- ions can be “exchanged" for molecules of the same charge. Thus, a cation exchange column will have positively charged counter-ions and positively charged compounds present in a mixture passed through the column will exchange with the counter-ions and “stick" to the negatively charged groups on the beads. Molecules in the sample that are neutral or negatively charged will pass quickly through the column. On the other hand, in anion exchange chromatography, the chemical groups attached to the beads are positively charged and the counter-ions are negatively charged. Molecules in the sample that are negatively charged will “stick" and other molecules will pass through quickly. To remove the molecules “stuck" to a column, one simply needs to add a high concentration of the appropriate counter-ions to displace and release them. This method allows the recovery of all components of the mixture that share the same charge.

Figure 9.3.1:  Cation exchange chromatography

Contributors

Dr. Kevin Ahern and Dr. Indira Rajagopal (Oregon State University)