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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

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Abstract

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.

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Authors and Affiliations

  1. Institute of Laboratory Medicine, Department of Clinical Chemistry & Transfusion Medicine, Sweden

    Annika E. Bäcker, Sara Thorbert, Eva C. Hallberg, Anne Olling, Mikael Gustavsson & Bo E. Samuelsson

  2. Bioenergetics Group Wallenberg Laboratory, Sahlgrenska universitetssjukhuset, SE 413 45, GÖTEBORG, Sweden

    Olivier Rakotonirainy & Bassam Soussi

Authors
  1. Annika E. Bäcker
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  2. Sara Thorbert
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  3. Olivier Rakotonirainy
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  4. Eva C. Hallberg
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  5. Anne Olling
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  6. Mikael Gustavsson
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  7. Bo E. Samuelsson
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  8. Bassam Soussi
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Correspondence to Annika E. Bäcker.

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Bäcker, A.E., Thorbert, S., Rakotonirainy, O. et al. 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. Glycoconj J 16, 45–58 (1999). https://doi.org/10.1023/A:1006901803636

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  • DOI: https://doi.org/10.1023/A:1006901803636

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