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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
10:21

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Published on: September 21, 2011

Contributions to reversed-phase column selectivity. I. Steric interaction.

P W Carr1, J W Dolan, U D Neue

  • 1University of Minnesota, Minneapolis, MN, USA.

Journal of Chromatography. A
|February 26, 2011
PubMed
Summary
This summary is machine-generated.

Steric interaction and shape selectivity in reversed-phase chromatography are distinct retention mechanisms. Steric interaction is driven by solute length, while shape selectivity is influenced by column properties and temperature.

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Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Reversed-phase chromatography (RPC) exhibits restricted retention for bulky solutes.
  • This restricted retention can manifest as either "shape selectivity" or "steric interaction."

Purpose of the Study:

  • To further examine the steric interaction process in RPC.
  • To compare steric interaction with shape selectivity.
  • To investigate the influence of column properties on these interactions.

Main Methods:

  • Analysis of data from 150 solutes and 167 monomeric type-B alkylsilica columns.
  • Determination of column hydrophobicity and steric interaction dependence on column properties (ligand length/concentration, pore diameter, end-capping).

Main Results:

  • Steric interaction is primarily dependent on solute molecular length; longer solutes exhibit greater steric interaction.
  • Steric interaction peaks with monomeric C(18) columns and decreases with C(1) or C(30) columns.
  • Shape selectivity is minimal for monomeric C(1)-C(18) columns at ambient/higher temperatures but significant for C(30) and polymeric columns (ligands ≥C(8)).

Conclusions:

  • Shape selectivity and steric interaction are affected differently by separation conditions and sample characteristics.
  • A hypothesis suggests shape selectivity involves cavity formation (enthalpic), while steric interaction utilizes pre-existing spaces (entropic).
  • The study also examined the dependence of hydrophobic interaction on column properties.