Size Exclusion Chromatography (SEC)

Abstract Size exclusion chromatography operates on a different principle than RPLC, IEC, and other chromatographic techniques. Individual gel permeation and gel filtration columns display a distribution of pore sizes. Polymers can only diffuse into pores that are larger than the polymer. The fraction, K, of pore volume accessible to the macromolecule of a given size is known as its permeation coefficient. Selectivity results from differing permeation coefficients. A column can be calibrated using well-characterized macromolecular standards. The calibration curve is of the form log(Mwt) vs. retention volume, and is typically used by software to automatically calculate the molecular weights of unknowns.

KeywordsGel filtration, Gel permeation, Polymers, Pores, Molecular weight, Macromolecule, Pore volume, Interstitial volume, Total exclusion, Total permeation, Permeation coefficient, Selective permeation, Deconvolution, Calibration curve


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