Related Experiment Video
Updated: Jul 5, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Impact of column temperature and mobile phase components on selectivity of hydrophilic interaction chromatography
Zhigang Hao1, Baiming Xiao, Naidong Weng
1Research and Development, Bristol-Myers Squibb Company, NJ, USA. zhigang.hao@gmail.com
Abstract:
The retention mechanism and chromatographic behavior for different polar analytes under hydrophilic interaction chromatography (HILIC) conditions have been studied by application of different mobile phases and stationary phases to various analytes at different temperatures. In addition to the commonly accepted mechanism of analyte liquid-liquid partitioning between mobile phase and water-enriched solvent layer which is partially immobilized onto the surface of the stationary phase, hydrogen-bonding, hydrophobic interaction, and ion-exchange interactions may also be involved. The predominant retention mechanism in HILIC separation is not always easily predictable. It can depend not only on the characteristics of the analytes but also on the selection of mobile and stationary phase compositions. The objective of this review is to evaluate the potential application of column temperature and mobile phase composition toward improving HILIC selectivity. The functional groups from analyte structures, stationary phase materials and organic mobile phase solvents will be highlighted.
Related Concept Videos
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
High-Performance Liquid Chromatography: Elution Process
Optimizing Chromatographic Separations
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
Ion-Exchange Chromatography
Silica Gel Column Chromatography: Overview
Polar components tend to bind strongly to the silica gel, causing them to move slowly through the column. In contrast, nonpolar compounds...
Gas Chromatography: Types of Columns and Stationary Phases
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...

