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Size Exclusion Chromatography

Size Exclusion Chromatography

Size Exclusion Chromatography (SEC) separates molecules based on their size, or more precisely, their hydrodynamic volume. It is based on the discrimination of individual sample components by the pores of the packing material. Large sample molecules cannot or can only partially penetrate the pores and elute from the column first, whereas smaller molecules can access all or a larger number of pores and elute later. SEC is the only mode of chromatography that does not involve interaction with a stationary phase by means of adsorption or partitioning of the solutes.

The terms SEC, GFC (gel filtration chromatography) and GPC (gel permeation chromatography) all refer to the same chromatographic technique. In GFC an aqueous mobile phase is used, while an organic mobile phase is employed in GPC. The general term SEC covers both uses.

Applications

SEC is the dominant mode of separation for natural and synthetic polymers: 

  • GFC is the term used for the size-based separation of  water-soluble polymers, for example biopolymers or natural polymers.

  • GPC is the term used for the size-based separation of polymers soluble in organic solvents.

TSKgel® SEC Columns

Size exclusion chromatography columns are traditionally packed with porous polystyrene divinylbenzene (PS-DVB) or silica particles.  PS-DVB columns are commonly used for the analysis of synthetic polymers in organic solvents, while silica-based columns are used for the separation of biopolymers.  


Aqueous SEC - Gel Filtration Chromatography

Gel Filtration Chromatography (GFC) is popular among biochemists for the isolation of proteins, for the removal of aggregates, to desalt a protein sample, to separate nucleic acid fractions, or to characterize water-soluble polymers used in food products, paints, pharmaceutical preparations, etc.


Organic SEC - Gel Permeation Chromatography 

Gel Permeation Chromatography (GPC) plays an important role in the characterization of industrial organic-soluble and polar organic-soluble polymers in the consumer, chemical and petrochemical industries.