Electrostatic repulsion hydrophilic interaction chromatography for isocratic separation of charged solutes and selective isolation of phosphopeptides

Anal Chem. 2008 Jan 1;80(1):62-76. doi: 10.1021/ac070997p. Epub 2007 Nov 21.

Abstract

If an ion-exchange column is eluted with a predominantly organic mobile phase, then solutes can be retained through hydrophilic interaction even if they have the same charge as the stationary phase. This combination is termed electrostatic repulsion-hydrophilic interaction chromatography (ERLIC). With mixtures of solutes that differ greatly in charge, repulsion effects can be exploited to selectively antagonize the retention of the solutes that normally would be the best retained. This permits the isocratic resolution of mixtures that normally require gradients, including peptides, amino acids, and nucleotides. ERLIC affords convenient separations of highly charged peptides that cannot readily be resolved by other means. In addition, phosphopeptides can be isolated selectively from a tryptic digest.

MeSH terms

  • Amino Acids / chemistry
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange / methods*
  • Hydrogen-Ion Concentration
  • Ion Exchange
  • Kinetics
  • Nucleic Acids / chemistry
  • Nucleotides / chemistry
  • Organic Chemicals / chemistry
  • Phosphopeptides / chemistry*
  • Phosphopeptides / isolation & purification*
  • Salts / chemistry
  • Solvents / chemistry
  • Static Electricity

Substances

  • Amino Acids
  • Nucleic Acids
  • Nucleotides
  • Organic Chemicals
  • Phosphopeptides
  • Salts
  • Solvents