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Hydrophilic-Interaction Chromatography in Peptide and Protein Analysis

Peptides and Proteins

  1. Paul Jenö

Published Online: 15 SEP 2006

DOI: 10.1002/9780470027318.a1619

Encyclopedia of Analytical Chemistry

Encyclopedia of Analytical Chemistry

How to Cite

Jenö, P. 2006. Hydrophilic-Interaction Chromatography in Peptide and Protein Analysis. Encyclopedia of Analytical Chemistry. .

Author Information

  1. Biozentrum of the University of Basel, Basel, Switzerland

Publication History

  1. Published Online: 15 SEP 2006


Hydrophilic-interaction liquid chromatography (HILIC) has been introduced as a highly efficient chromatographic technique for the separation of a wide range of solutes. A hydrophilic stationary phase is eluted with a hydrophobic mobile phase whereby retention of solutes increases with the hydrophilicity of solutes. Bound solutes are eluted by decreasing the hydrophobicity of the mobile phase. Mixed mode effects based on hydrophilic and ionic interactions can also be exploited by proper selection of the stationary and the mobile phase. Typically, the order of elution is the opposite of that obtained by reversed-phase chromatography (RPC). HILIC is particularly interesting for solutes which are difficult to chromatograph on conventional reversed-phase materials. This review focuses on the application of HILIC to the separation of amphipathic α-helical peptides, the analysis of protein glycosylation, phosphorylation, and acetylation, and to the removal of detergents from membrane proteins.