chip-based amide-HILIC LC/MS platform for glycosaminoglycan glycomics profiling

A key challenge to investigations into the functional roles of glycosaminoglycans (GAGs) in biological systems is the difficulty in achieving sensitive, stable, and reproducible mass spectrometric analysis. GAGs are linear carbohydrates with domains that vary in the extent of sulfation, acetylation,...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2009-02, Vol.9 (3), p.686-695
Main Authors: Staples, Gregory O, Bowman, Michael J, Costello, Catherine E, Hitchcock, Alicia M, Lau, James M, Leymarie, Nancy, Miller, Christine, Naimy, Hicham, Shi, Xiaofeng, Zaia, Joseph
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Carbohydrates
Cattle
Chromatography, Liquid - methods
Complex mixture identification
Electrospray ionization‐quadrupole time of flight tandem mass spectrometry
Fundamental and applied biological sciences. Psychology
Glycomics
Glycomics - methods
Glycosaminoglycans - analysis
Heparitin Sulfate - chemistry
Hydrophobic and Hydrophilic Interactions
Miscellaneous
Proteins
Spectrometry, Mass, Electrospray Ionization - methods
Swine
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Summary:A key challenge to investigations into the functional roles of glycosaminoglycans (GAGs) in biological systems is the difficulty in achieving sensitive, stable, and reproducible mass spectrometric analysis. GAGs are linear carbohydrates with domains that vary in the extent of sulfation, acetylation, and uronic acid epimerization. It is of particular importance to determine spatial and temporal variations of GAG domain structures in biological tissues. In order to analyze GAGs from tissue, it is useful to couple MS with an on-line separation system. The purposes of the separation system are both to remove components that inhibit GAG ionization and to enable the analysis of very complex mixtures. This contribution presents amide-silica hydrophilic interaction chromatography (HILIC) in a chip-based format for LC/MS of heparin, heparan sulfate (HS) GAGs. The chip interface yields robust performance in the negative ion mode that is essential for GAGs and other acidic glycan classes while the built-in trapping cartridge reduces background from the biological tissue matrix. The HILIC chromatographic separation is based on a combination of the glycan chain lengths and the numbers of hydrophobic acetate (Ac) groups and acidic sulfate groups. In summary, chip based amide-HILIC LC/MS is an enabling technology for GAG glycomics profiling.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200701008