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Updated: May 21, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Identifying orthogonal and similar reversed phase liquid chromatography stationary phases using the system
Andrew R Johnson1, Carrie M Johnson, Dwight R Stoll
1Indiana University, Department of Chemistry, 800 E. Kirkwood Ave., Bloomington, IN 47405, USA.
Abstract:
We have compared over 500 RPLC columns characterized by the hydrophobic subtraction model using the system selectivity cube (SSC). We have shown numerous differences in column selectivity even among columns in the same class (e.g., alkyl-silica, cyano, or embedded polar groups). We also illustrate the utility of our method for selecting alternative columns with different selectivities for problematic separations and for selecting orthogonal columns for use in two-dimensional separations. The system selectivity cube offers a visual way to easily compare many columns simultaneously and select those columns offering the desired selectivity.
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