Towards a more complete glycome: Advances in ion chromatography-mass spectrometry (IC-MS) for improved separation and analysis of carbohydrates

J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jun 15:1175:122719. doi: 10.1016/j.jchromb.2021.122719. Epub 2021 Apr 20.

Abstract

To date, few tools are available for the analysis of the glycome without derivatization, a process which is known to introduce issues such as differential loss of sialic acid and incomplete labeling. We have previously reported the use of ion chromatography-mass spectrometry (IC-MS) to analyze native sialylated and sulfated glycans. Here, we introduce improvements to IC column technology, enabling the separation of neutral glycans while maintaining charge separation capabilities. When implemented in an IC-MS workflow, this enables the structural characterization of a broad array of chemically distinct glycans. With the newly developed IC column and modified IC-MS instrumentation configuration, we qualitatively investigated O-glycome profiles in bovine fetuin and porcine gastric mucins. The improved chromatographic resolution in combination with high-resolution MS data present a powerful tool for glycan structural identification.

Keywords: Anion exchange chromatography; Carbohydrate; Derivatization-free; Glycomics; HPAE-MS; Pulsed amperometric detection.

MeSH terms

  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid
  • Fetuins / chemistry
  • Glycomics / methods*
  • Milk / chemistry*
  • Mucins / chemistry
  • N-Acetylneuraminic Acid / analysis
  • Polysaccharides / analysis*
  • Sulfates / analysis
  • Swine
  • Tandem Mass Spectrometry

Substances

  • Fetuins
  • Mucins
  • Polysaccharides
  • Sulfates
  • N-Acetylneuraminic Acid