Liquid chromatography "on-flow" 1H nuclear magnetic resonance on native glycosphingolipid mixtures together with gas chromatography/mass spectrometry on the released oligosaccharides for screening and characterisation of carbohydrate-based antigens from pig lungs.
1999
Glycosphingolipids were prepared from pig lung and pooled into two fractions with (i) ≥3 sugar residues, and (ii) ≤3 sugar residues. Oligosaccharides were prepared and used for gas chromatography, gas chromatography-mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. The glycolipid fractions i and ii were further characterised and purified using a novel method based on high performance liquid chromatography “on-flow” proton nuclear magnetic resonance. The LC “on-flow” NMR technique showed good chromatographic separation and gave NMR spectral information which could be used as guidance for pooling of the separated mixture glycolipids. Conventional 1H NMR, thin layer immunostaining, gas chromatography, gas chromatography/mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry were used to characterise the glycolipids and to validate LC-NMR spectral data.
Keywords:
- Sample preparation in mass spectrometry
- Hydrophilic interaction chromatography
- High-performance liquid chromatography
- Liquid chromatography–mass spectrometry
- Gas chromatography
- Protein mass spectrometry
- Mass spectrometry
- Direct electron ionization liquid chromatography–mass spectrometry interface
- Chromatography
- Analytical chemistry
- Chemistry
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