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Discrimination of sulfated isomers of chondroitin sulfate disaccharides by HILIC-MS

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Abstract

Chondroitin sulfate (CS) glycosaminoglycans are biologically active sulfated polysaccharides that pose an analytical challenge for their structural analysis and functional evaluation. In this study, we developed a hydrophilic interaction liquid chromatography separation method and its on-line coupling to mass spectrometry (MS) allowing efficient differentiation and sensitive detection of mono-, di-, and trisulfated CS disaccharides and their positional isomers, without requiring prior derivatization. The composition of the mobile phase in terms of pH and concentration showed great influence on the chromatographic separation and was varied to allow the distinction of each CS without signal overlap for a total analysis time of 25 min. This methodology was applied to determine the disaccharide composition of biological reaction media resulting from various enzymatic transformations of CS, such as enzymatic desulfation of CS disaccharides by a CS 4-O-endosulfatase, and depolymerization of the CS endocan by chondroitinase lyase ABC.

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Acknowledgements

We would like to thank the LabEx CHARMMMAT (ANR-11-LABX-0039) for the attribution of a post-doctoral fellowship (SP) and the partial funding for the acquisition of the Bruker AMAZON SPEED ETD ion trap. IS acknowledges PhD fellowships from doctoral school SDSV (N°577, Université Paris-Saclay).

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Authors and Affiliations

  1. Université Paris-Saclay, CNRS, Univ Evry, LAMBE, 91025, Evry-Courcouronnes, France

    Salomé Poyer, Ilham Seffouh, Jean-Yves Salpin & Régis Daniel

  2. CY Cergy Paris Université, LAMBE, 91025, Evry-Courcouronnes, France

    Salomé Poyer, Ilham Seffouh, Jean-Yves Salpin & Régis Daniel

  3. ICOA, CNRS UMR 7311, Université d’Orléans, 45067, Orléans, France

    Chrystel Lopin-Bon & Jean-Claude Jacquinet

  4. Instituto de Biología Molecular y Celular, Universidad Miguel Hernandez, Elche, Alicante, Spain

    José L. Neira

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  1. Salomé Poyer
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  2. Ilham Seffouh
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Correspondence to Salomé Poyer or Régis Daniel.

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Poyer, S., Seffouh, I., Lopin-Bon, C. et al. Discrimination of sulfated isomers of chondroitin sulfate disaccharides by HILIC-MS. Anal Bioanal Chem 413, 7107–7117 (2021). https://doi.org/10.1007/s00216-021-03679-9

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  • DOI: https://doi.org/10.1007/s00216-021-03679-9

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