ION EXCHANGE CHROMATOGRAPHY

ION EXCHANGE CHROMATOGRAPHY

Separates & purifies charged molecules based on their net charge.

Relies on the reversible exchange of ions between the sample molecules and charged groups immobilised on a stationary phase.

Stationary Phase: The column is packed with a matrix containing charged functional groups, either positively charged (cation exchange) or negatively charged (anion exchange).

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Sample Loading: The sample containing charged molecules is applied to the column. Positively charged molecules will bind to

negatively charged stationary phase (anion

exchange), and negatively charged

molecules will bind to positively charged

stationary phase (cation exchange).

Washing: Unbound molecules and

impurities are washed through the column using a buffer solution that doesn't disrupt the binding interaction.

To

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Elution: Elution is done by changing the ionic strength or pH of the buffer. This disrupts the electrostatic interactions between the sample molecules and the stationary phase, releasing the bound molecules.

Collecting Fractions: As different

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molecules have different affinities for the

stationary phase, they will elute at

different times, resulting in separation. Fractions containing the separated components can be collected.