Polysaccharides Structure Analysis

  • Glycosyl composition – We can provide qualitative and quantitative monosaccharide composition analysis. At least 100-500 micrograms of sample is used for this analysis.
  • Glycosyl linkage We apply methods of partially methylated alditol acetates (PMAA) with additional modifications required for determination of uronic acids or unusual carbohydrate substituents.  200mg up to 1mg or higher amounts of sample is required for linkage analysis.
  • Molecular weight determination Size Exclusion Chromatography (SEC). SEC analysis provides information on molecular weight distribution of complex polysaccharide or oligosaccharide mixtures. 200mg-1mg or higher amounts of sample is required for linkage analysis.
  • 1D- and 2D-NMR-spectroscopy 1D (1H, 13C) and standard 2D homo- and heteronuclear  nuclear magnetic resonance (including inter alia 1H-1H-COSY, TOCSY, NOESY, ROESY, 1H-13C-HSQC, H2BC, HSQC-TOCSY, and HMBC) is an extremely powerful technique for determination of carbohydrate sequence and structure of plant poly- and oligosaccharides. It also helps to evaluate sample quality, – α/β – anomeric configuration of glycosyl residues, and quick assessment of the nature of purified polysaccharide.  Information on sample requirements can be found at: NMR Facilities
  • Enzymatic digestion Depolymerization or cleavage of plant polysaccharides helps to  generate oligosaccharides suitable for structural characterization by mass-spectrometry (MALDI-TOF MS or ESI-MS) and 1D and 2D NMR-spectroscopy.
  • High-Performance – Anion Exchange Chromatography (HPAEC)  – Separation of plant derived oligosaccharides based on their mass to charge ratio allow identification and quantification of lower oligomers (up to DP8-10) based on availability  appropriate reference standards.
  • Determination of absolute configuration of glycosyl residues – ‘D’ and ‘L’ absolute configuration of individual glycosyl residues can be determined by acid-catalyzed derivatization with optically pure 2-butanol, followed by per-O-trimethylsilylation and GC-MS analysis. Identification of the D and L forms is made by comparison of retention times the standard D or L monosaccharides. A minimum of 200-500 micrograms of carbohydrate is required, more if available.
  • Plant Cell Wall Glycome Profiling by high throughput ELISA automated system – Glycome profiling, which is an immunological technique for rapid characterization of plant cell wall/biomass samples that was developed in Dr. Michael Hahn’s laboratory at the CCRC. This semi-quantitative analytical technique takes advantage of a large and diverse toolkit of monoclonal antibodies (mAbs) directed at each of the major non-cellulosic plant cell wall polysaccharides, except rhamnogalacturonan II. The mAb collection includes multiple antibodies against epitopes on xyloglucans, xylans, homogalacturonans, rhamnogalacturonan I, mannans, and arabinogalactans.