Size‐exclusion Chromatography Gradients

Author:

Radke Wolfgang

Abstract

AbstractGradient chromatography is an established technique for the separation of polymers according to chemical composition. Gradient separation involves injection of the dissolved polymer sample into a chromatographic column at conditions such that the sample is either adsorbed or precipitated at the column head. Afterward, the sample components are successively desorbed or redissolved and eluted from the column by changing the eluent composition in a programmed manner. However, polymers are usually soluble only in a low number of solvents. Therefore, the solvent applied for dissolving the material often prevents adsorption to the stationary phase. This results in nonretained sample components eluting with the actual injected solvent in the so‐called breakthrough peaks. Breakthrough peaks might occur as well in gradient separations based on solubility differences, when the sample solvent is strong enough to prevent the desired precipitation on the column head. These breakthrough peaks hamper method development and quantitation of the different components. Size‐exclusion chromatography (SEC) gradients provide an alternative way of performing gradient separations of high molar mass samples without the risk of breakthrough peaks. In contrast to conventional gradients in SEC gradients, the sample is dissolved in a solvent that acts as strong eluent for the sample components, allowing in principle using the commonly applied SEC solvents as starting points. This article explains the principles of SEC gradients and provides examples, showing the potential and limitations of this rather new approach.

Publisher

Wiley

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