Off-line coupling of capillary isotachophoresis separation to IRMPD spectroscopy for glycosaminoglycans analysis: Application to the chondroitin sulfate disaccharides model solutes
| Autor: | Isabelle Compagnon, Baptiste Schindler, Jérôme Randon, Léna Jaravel, Vincent Dugas, Claire Demesmay |
|---|---|
| Přispěvatelé: | Separative Methods - Techniques séparatives, Institut des Sciences Analytiques (ISA), Institut de Chimie du CNRS (INC)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Structure et dynamique multi-échelle des édifices moléculaires (DYNAMO), Institut Lumière Matière [Villeurbanne] (ILM), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Formic acid
Electrolyte Fractionation 010402 general chemistry Mass spectrometry Disaccharides 01 natural sciences Biochemistry Dissociation (chemistry) Analytical Chemistry Capillary electrophoresis chemistry.chemical_compound Electrolytes Tandem Mass Spectrometry [CHIM.ANAL]Chemical Sciences/Analytical chemistry [CHIM]Chemical Sciences [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Infrared multiphoton dissociation Glycosaminoglycans Chromatography Isotachophoresis Chemistry Spectrum Analysis 010401 analytical chemistry Organic Chemistry Chondroitin Sulfates Electrophoresis Capillary General Medicine Microscale separation 0104 chemical sciences Feasibility Studies Off-line coupling |
| Zdroj: | Journal of Chromatography A Journal of Chromatography A, Elsevier, 2020, 1617, pp.460782. ⟨10.1016/j.chroma.2019.460782⟩ |
| ISSN: | 0021-9673 1873-3778 |
| Popis: | International audience; Glycans analysis is challenging due to their immense structural diversity. Isotachophoresis was investigated as separation method for the purification of isobaric sulfated disaccharides prior to their characterization by Mass Spectrometry (MS) and tunable IR multiple photon dissociation (IRMPD). This proof of feasibility study was applied to the separation and characterization of chondroitin sulfate (CS) disaccharides. ITP separation conditions were optimized. Separation starts using a 2.5 mM chloride ions and 10 mM glycine at pH 3.2 solution as leading electrolyte and a terminating electrolyte composed of formic acid 2.5 mM and glycine 10 mM at pH 3.5. The CS disaccharides sample were prepared in the terminating electrolyte. The length of injection was also investigated in order to create longer plateau-like bands of pure solutes. This strategy was helpful for collecting fraction at such microseparation scale. Indeed, capillary ITP affords the injection of few tens of nanoliter of sample. Fractionation of the CS disaccharides mixture in isolated ITP bands and collection of solutes were successfully done using a HPC coated fused silica capillary of 1m-length and 75 µm of internal diameter. Collected fractions in a final of volume 10 µL were analyzed by CZE, tandem MS and IRMPD spectroscopy. The purity of each fraction is higher than 90% and is well-adapted to IRMPD characterization. |
| Databáze: | OpenAIRE |
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