Sugar Chain Analysis by Enzymatic Digestion and 2D Mapping by HPLC

  • Megumi Maeda
  • Yoshinobu Kimura


Structural analysis is crucial to elucidating the physiological functions of oligosaccharides. The rapid development of both hardware and software for NMR spectrometry and mass spectrometry (ESI-MS and MALDI-MS) has enabled significantly faster and easier structural analysis. When analyzing a small amount of oligosaccharide, the combined use of endo- and exo-glycosidases having clear-cut substrate specificities provides large and reliable advantages for structural analysis. For structural analysis using glycosidases, which are used for high-sensitivity detection, oligosaccharides are usually labeled with tritium or fluorescence reagents: 2-aminopyridine (Hase et al. 1978, 1987) or 2-aminobenzamide (Rudd et al. 1997) in advance. When fluorescent reagents are used for labeling oligosaccharides, reverse-phase HPLC can be used for structural analysis because of the increase in the hydrophobicity of sugar chains. Therefore, two-dimensional (2D)-HPLC using both reverse-phase and normal-phase columns can be used for identifying oligosaccharide structures. 2D-HPLC can distinguish the structural difference (anomeric or branching structures) between isomeric oligosaccharides of the same molecular size or sugar composition using a small amount of sample. Here, as a typical example, we describe the structural analysis of a novel N-glycan obtained from royal jelly glycoproteins using fluorescence labeling and 2D-HPLC developed by Hase et al. (1978, 1987).


Sodium Acetate Buffer Sugar Chain Royal Jelly Jack Bean Elution Position 
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Copyright information

© Springer 2008

Authors and Affiliations

  • Megumi Maeda
    • 1
  • Yoshinobu Kimura
    • 2
  1. 1.Department of HygieneKawasaki Medical SchoolKurashikiJapan
  2. 2.Department of Biofunctional Chemistry, Graduate School of Natural Science and TechnologyOkayama UniversityOkayamaJapan

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