On-line selective enrichment and ion-pair reaction for structural determination of sulfated glycopeptides by capillary electrophoresis-mass spectrometry

J Chromatogr A. 2008 Jun 20;1194(2):237-42. doi: 10.1016/j.chroma.2008.04.050. Epub 2008 Apr 24.

Abstract

We describe a new capillary electrophoresis-mass spectrometry (CE-MS)-based technique for analyzing sulfated glycopeptides. The proposed method performs selective enrichment of sulfated glycopeptides from a complex mixture of peptides based on field-enhanced sample injection and ion-pair reaction with a basic ion-pair reagent (Lys-Lys-Lys; KKK) at the exit end of a capillary in a single analysis, which permits successful fragmentation of sulfated glycopeptides in positive-ion mode at the MS/MS stage for comprehensive structural analysis. In this study, the method was verified using a model sulfated monosaccharide, N-acetyl-d-galactosamine 4-sulfate (GalNAc 4S). As an example of an application of this method, sulfated glycopeptides were selectively enriched from the enzymatic digest of thyroid stimulating hormone, affording approximately 500-fold sensitivity enhancement, and structural information was successfully obtained via on-line ion-pair complexation reaction.

MeSH terms

  • Electrophoresis, Capillary / instrumentation
  • Electrophoresis, Capillary / methods*
  • Glycopeptides / analysis*
  • Glycopeptides / chemistry
  • Mass Spectrometry / instrumentation
  • Mass Spectrometry / methods*
  • Molecular Structure
  • Reproducibility of Results
  • Sulfuric Acid Esters / analysis*
  • Sulfuric Acid Esters / chemistry

Substances

  • Glycopeptides
  • Sulfuric Acid Esters