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In size exclusion chromatography (SEC) separation is not based on interactions between the analyte and the column surface; separation relies on the degree of inclusion or exclusion of an analyte from the pores within the stationary phase. The stationary phase pores have different accessibility for molecules depending on their size (hydrodynamic volume), i.e. larger molecules are generally poorly retained and elute first as they cannot move into the pores within the stationary phase, whereas smaller molecules can diffuse into the pores to a greater extent, depending on their size, and will therefore be eluted later, with the smallest molecules eluting last. This module focuses on size exclusion chromatography for the analysis of biomolecules.


Topics include:

  • Basics of size exclusion chromatography (SEC)
  • Size exclusion chromatography calibration curves
  • Size exclusion chromatography for biomolecules
  • Stationary phases in size exclusion chromatography
  • Mobile phase and secondary interactions in size exclusion chromatography
  • Instrument considerations
  • Modern approaches to size exclusion chromatography
  • Selection of columns and conditions for bio size exclusion chromatography