Gas-phase pyridylamination of saccharides: development and applications

Anal Chem. 2007 Apr 1;79(7):2674-9. doi: 10.1021/ac0700878. Epub 2007 Mar 3.

Abstract

Pyridylamination is a versatile method for fluorescence labeling of oligosaccharides. The technique affords sensitive detection of saccharides with reducing termini and high-resolution separation by high-performance liquid chromatography. The conventional method, based on a liquid-phase reaction, has been extensively used in various aspects of glycobiology and glycotechnology. Unfortunately, the necessity for removing excess 2-aminopyridine makes the technique both laborious and time-consuming. Furthermore, removal of excess reagent can result in a significant loss of short saccharide components. In the present paper, we report an alternative methodology based on a "gas-phase" reaction, in which dried saccharides are reacted with vaporized 2-aminopyridine. The resultant Schiff base was also reduced in the gas phase within the same reaction microtube using a purpose-built device. The newly developed procedure was applied to both monosaccharide (GlcNAc) and oligosaccharides (isomalto-oligosaccharides) at quantitative yields with no requirement to remove excess reagent. The acid-labile sialyl linkages of alpha2-6-disialobiantennary oligosaccharides proved to be fully stable during the procedure. The developed method was also successfully applied to profiling N-linked oligosaccharides liberated from glycoproteins by hydrazinolysis and, thus, should contribute to various fields of glycomics.

MeSH terms

  • Aminopyridines / chemistry*
  • Chromatography, High Pressure Liquid / methods
  • Gases / chemistry
  • Oligosaccharides / chemistry*
  • Oligosaccharides / isolation & purification
  • Schiff Bases / chemistry
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Aminopyridines
  • Gases
  • Oligosaccharides
  • Schiff Bases
  • alpha-aminopyridine