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Updated: Jun 28, 2026
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Mass Spectrometric Analysis of Glycosphingolipid Antigens
Published on: April 16, 2013
Structural glycomics using hydrophilic interaction chromatography (HILIC) with mass spectrometry
Manfred Wuhrer1, Arjen R de Boer, André M Deelder
1Leiden University Medical Center, Biomolecular Mass Spectrometry Unit, Department of Parasitology, P.O. Box 9600, 2300 RC Leiden, The Netherlands. m.wuhrer@lumc.nl
Mass Spectrometry Reviews
|November 4, 2008
Summary
Hydrophilic interaction chromatography (HILIC) coupled with mass spectrometry offers powerful glycomics analysis. This technique efficiently separates glycans and glycopeptides, even resolving complex isobaric species with high sensitivity.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Glycomics
Background:
- Structural glycomics is crucial for understanding biological processes.
- Glycans and glycopeptides require advanced separation techniques for analysis.
- Mass spectrometry provides sensitive detection for biomolecules.
Purpose of the Study:
- To highlight the utility of hydrophilic interaction chromatography (HILIC) for glycomics.
- To detail the separation mechanisms and applications of HILIC-MS.
- To compare online HILIC-MS with offline MALDI-MS(/MS) for glycan analysis.
Main Methods:
- Hydrophilic interaction chromatography (HILIC) utilizing hydrogen bonding, ionic, and dipole-dipole interactions for glycan retention.
- Online coupling of HILIC with nano-scale electrospray ionization-mass spectrometry (ESI-MS).
- Offline analysis of glycans using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS(/MS)) in both negative and positive ion modes.
Main Results:
- HILIC-MS enables efficient separation of glycans, glycopeptides, and modified glycans, resolving isobaric species.
- Nano-scale HILIC-MS achieves a detection limit of approximately 1 femtomol for oligosaccharides.
- MALDI-MS(/MS) provides informative fragment ion spectra for deprotonated species (negative mode) and sodium adducts (positive mode).
Conclusions:
- HILIC coupled with mass spectrometry is a versatile and sensitive technique for comprehensive structural glycomics.
- Both online HILIC-MS and offline MALDI-MS(/MS) are valuable complementary methods for detailed glycan structural elucidation.
- The described methods facilitate high-resolution separation and sensitive detection of diverse glycan structures.
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