Introducing porous graphitized carbon liquid chromatography with evaporative light scattering and mass spectrometry detection into cell wall oligosaccharide analysis

Y. Westphal, H.A. Schols, A.G.J. Voragen, H. Gruppen

Research output: Contribution to journalArticleAcademicpeer-review

34 Citations (Scopus)

Abstract

Separation and characterization of complex mixtures of oligosaccharides is quite difficult and, depending on elution conditions, structural information is often lost. Therefore, the use of a porous-graphitized-carbon (PGC)-HPLC-ELSD-MSn-method as analytical tool for the analysis of oligosaccharides derived from plant cell wall polysaccharides has been investigated. It is demonstrated that PGC-HPLC can be widely used for neutral and acidic oligosaccharides derived from cell wall polysaccharides. Furthermore, it is a non-modifying technique that enables the characterization of cell wall oligosaccharides carrying, e.g. acetyl groups and methylesters. Neutral oligosaccharides are separated based on their size as well as on their type of linkage and resulting 3D-structure. Series of the planar ß-(1,4)-xylo- and ß-(1,4)-gluco-oligosaccharides are retained much more by the PGC material than the series of ß-(1,4)-galacto-, ß-(1,4)-manno- and a-(1,4)-gluco-oligosaccharides. Charged oligomers such as a-(1,4)-galacturonic acid oligosaccharides are strongly retained and are eluted only after addition of trifluoroacetic acid depending on their net charge. Online-MS-coupling using a 1:1 splitter enables quantitative detection of ELSD as well as simple identification of many oligosaccharides, even when separation of oligosaccharides within a complex mixture is not complete. Consequently, PGC-HPLC-separation in combination with MS-detection gives a powerful tool to identify a wide range of neutral and acidic oligosaccharides derived from various cell wall polysaccharides.
Original languageEnglish
Pages (from-to)689-695
JournalJournal of Chromatography. A, Including electrophoresis and other separation methods
Volume1217
Issue number5
DOIs
Publication statusPublished - 2010

Keywords

  • treated eucalyptus wood
  • hairy ramified regions
  • capillary-electrophoresis
  • black-currants
  • polysaccharides
  • pectin
  • xylogalacturonan
  • quantification
  • nomenclature
  • ionization

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