Principles of Size Exclusion Chromatography SEC (GFC, GPC)

Basics

Size Exclusion Chromatography (SEC) separates analytes by their molecular weight (MW) and/or the hydrodynamic radius (size, shape of the molecule).

Hydrodynamic radius = variable size of an analyte with specific MW based on dynamical properties

The packed columns have a stationary phase with defined pore size suitable for a range of molecular weight. Analytes and stationary phase with defined pore size

The smallest compounds in the sample mixture can enter deep into the pores and have a long elution time. The medium-sized components penetrate the pores in some depth and elute separately according to their size. Macromolecules, that are bigger than the pore size will not be retained. The maximum molecular weight that can enter the pore is called exclusion limit. exclusion limit (no interaction with pores, molecules too large to be trapped)

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