Skip to main content
SpringerLink
Log in
Book cover

Capillary Electrophoresis of Carbohydrates pp 41–69Cite as

Derivatization of Carbohydrates

  • Protocol

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 213))

Abstract

The monosaccharides comprise a number of isomers and homologues having very similar structures, basically composed of hydroxymethyl or hydroxymethine groups linked to each other. In reducing monosaccharides the hemiacetal group is at the reducing ends in addition to these polyhydroxylated basal structures. There are other series of monosaccharides with the ketal or carboxyl group instead of or in addition to the hemiacetal group. Substitution of the hydroxyl group by the hydrogen atom or the amino group produces the deoxy sugar or the amino sugar, respectively. All these functional groups give monosaccharides hydrophilic properties. With the exception of monosaccharide residues containing a carboxyl group, detection of these compounds following chromatographic and electrophoretic separation is rendered difficult owing to their inherent lack of chromophores in the ultraviolet (UV) and visible spectrum regions. Obviously, oligosaccharides and polysaccharides composed of these monosaccharide units have a similar shortcoming with respect to their detection.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Honda, S., Iwase, S., Makino, A., and Fujiwara, S. (1989) Simultaneous determination of reducing monosaccharides by capillary zone electrophoresis as the borate complexes of N-2-pyridylamines. Analyt. Biochem. 176, 72–77.

    Article  PubMed  CAS  Google Scholar 

  2. Grill, E., Huber, C., Oefner, P., Vorndran, A., and Bonn, G. K. (1993) Capillary zone electrophoresis of p-aminobenzoic acid derivatives of aldoses, ketoses, and uronic acids. Electrophoresis 14, 1004–1010.

    Article  PubMed  CAS  Google Scholar 

  3. Vorndran, A. E., Grill, E., Huber, C., Oefner, P. J., and Bonn, G. K. (1992) Capillary zone electrophoresis of aldoses, ketoses and uronic acids derivatized with ethyly p-aminobenzoate. Chromatographia 34, 109–114.

    Article  CAS  Google Scholar 

  4. Schwaiger, H., Oefner, P. J., Hubber, C., Grill, E., and Bonn, G. K. (1994) Capillary zone electrophoresis and micellar electrokinetic chromatography of 4-aminobneonitrile carbohydrate derivatives. Electrophoresis 15, 941–952.

    Article  PubMed  CAS  Google Scholar 

  5. Plocek, J. and Novotny, M. V. (1997) Capillary zone electrophoresis of oligosaccharides derived with N-(4-aminobenzoyl)-l-glutamic acid for ultraviolet absorbance detection. J. Chromatogr. A 757, 215–223.

    Article  PubMed  CAS  Google Scholar 

  6. Nashabeh, W. and El Rassi, Z. (1992) Capillary zone electrophoresis of linear and branched oligosaccharides. J. Chromatogr. A 600, 279–287.

    Article  CAS  Google Scholar 

  7. Camilleri, P., Harland, G. B., and Okafo, G. (1995) High resolution and rapid analysis of branched oligosaccharides by capillary electrophoresis. Analyl. Biochem. 230, 115–122.

    Article  CAS  Google Scholar 

  8. Mort, A. J. and Chen, E. M. W. (1996) Separation of 8-aminonaphthalene-1,3,6-trisulfonate (ANTS)-labeled oligomers containing galacturonic acid by capillary electrophoresis: application to determining the substrate specificity of endopolygalaturonases. Electrophoresis 205, 108–114.

    Google Scholar 

  9. Lee, K. B., Desai, U. R., Palcic, M. M., Hindsugaul, O., and Linhardt, R. J. (1992) An electrophoresis-based assay for glycosyltransferase activity. Analyl. Chem. 205, 108–114.

    CAS  Google Scholar 

  10. Chiesa, C. and Horváth, C. (1993) Capillary zone electrophoresis of linear and branched oligosaccharides. J. Chromatogr. A 645, 337–352.

    Article  CAS  Google Scholar 

  11. Guttman, A., Chen, F. A., Evangelista, R. A., and Cooke, N. (1996) High-resolution capillary gel electrophoresis of reducing oligosaccharides labeled with 1-aminopyrene-3,6,8-tirsulfonate. Analyt. Biochem. 233, 234–242.

    Article  PubMed  CAS  Google Scholar 

  12. Liu, J., Shirota, O., and Novotny, M. (1991) Ultrasensitive fluorometric detection of carbohydrates as derivatives in mixture separated by capillary electrophoresis. Proc. Natl. Acad. Sci. USA 8, 2302–2306.

    Article  Google Scholar 

  13. Honda, S., Okeda, J., Iwanaga, H., Taga, A., Suzuki, S., and Imai, K. (2000) Ultramicro analysis of reducing carbohydrates by capillary electrophoresis with laser-induced fluorescence detection as 7-nitro-2,1,3-benxoxadiazole-tagged N-methylglycamine derivatives. Analyt. Biohcem. 286, 99–111.

    Article  CAS  Google Scholar 

  14. Honda, S., Suzuki, S., Nose, A., Yamamoto, K., and Kakehi, K. (1991) Capillary zone electrophoresis of reducing mono-and oligosaccharides as borate complexes of their 3-methyl-1-phenyl-2-pyrazolin-5-one derivatives. Carbohydr. Res. 215, 193–198.

    Article  CAS  Google Scholar 

  15. Honda, S., Akao, E., Okuda, M., Suzuki, S., Kakehi, K., and Nakamura, J. (1989) High performance liquid chromatography of reducing carbohydrates as strongly ultraviolet absorbing and electrochemically active 1-phenyl-3-methyl-5-pyrazolone derivatives. Analyt. Biochem. 180, 351–357.

    Article  PubMed  CAS  Google Scholar 

  16. Suzuki, S., Tanaka, R., Takada, K., Inoue, N., and Honda, S. (2001) Analysis of sialo-N-glycans in glycoproteins as 1-phenyl-3-methyl-5-pyrazolone derivatives by capillary electrophoresis. J. Chromatogr. A 910, 319–329.

    Article  PubMed  CAS  Google Scholar 

  17. Honda, S., Ueno, T., and Kakehi, K. (1992) High performance capillary electrophoresis of unsaturated oligosaccharides derived from glycosaminoglycans by digestion with chondroitinase ABC as 1-phenyl-3-methyl-5-pyrazolone derivatives. J. Chromatogr. 608, 289–295.

    Article  PubMed  CAS  Google Scholar 

  18. Honda, S., Togashi, K., and Taga, A. (1997) Unusual separation of 1-phenyl-3-methyl-5-pyrazolone derivatives of aldoses by capillary zone electrophoresis. J. Chromatogr. A 791, 307–311.

    Article  CAS  Google Scholar 

  19. Honda, S., Yamamoto, K., Suzuki, S., Ueda, M., and Kakehi, K. (1991) High performance capillary zone electrophoresis of carbohydrates in the presence of alkaline earth metal ions. J. Chromatogr. 588, 327–333.

    Article  CAS  Google Scholar 

  20. Chiesa, C., Oefner, P. J., Zieske, L. R., and O’Neill, R. A. (1995) Micellar electrokinetic chromatography of monosaccharides derivatized with 1-phenyl-3-methyl-2-pyrazolin-5-one. J. Capil. Electrophor. 2, 175–183.

    CAS  Google Scholar 

  21. Honda, S., Togashi, K. Uegaki, K., and Taga, A. (1998) Enhancement of the zone electrophoretic separation of 1-phenyl-3-methyl-5-pyrazolone derivatives of aldoses as borate complexes by concerted ion-interaction electrokinetic chromatography with Polybrene. J. Chromatogr. A 805, 277–284.

    Article  CAS  Google Scholar 

  22. Taga, A., Yabusako, Y., Kitano, A., and Honda, S. (1998) Separation of oligosaccharides by affinity capillary electrophoresis. Electrophoresis 19, 2645–2649.

    Article  PubMed  CAS  Google Scholar 

  23. Honda, S., Taga, A., Kotanim M., and Grover, E. R. (1997) Separation of aldose enantiomers by capillary electrophoresis in the presence of optically active N-dodecoxycarbonylvalines. J. Chromatogr. A 792, 385–391.

    Article  CAS  Google Scholar 

  24. Suzuki, S., Kakehi, K., and Honda, S. (1996) Comparison of the sensitivities of various derivatives of oligosaccharides in LC/MS with fast atom bombardment and electrospray ionization interfaces. Analyt. Chem. 68, 2073–2083.

    Article  CAS  Google Scholar 

  25. Wang, W. T., LeDonne, N. C., Ackerman, B., and Sweeley, C. C. (1984) Structural characterization of oligosaccharides by high-performance liquid chromatography, fast-atom bombardment-mass spectrometry, and exoglycosidase digestion. Analyt. Biochem. 141, 366–381.

    Article  PubMed  CAS  Google Scholar 

  26. Hase, S., Hara, S., and Matsushima, Y. (1979) Tagging of sugars with a fluorescent compound, 2-aminopyridine. J. Biochem. 85, 217–220.

    PubMed  CAS  Google Scholar 

  27. Jackson, P. (1990) The use of polyacrylamide-gel electrophoresis for the highresolution separation of reducing saccharides labelled with the fluorophore 8-aminonaphthalene-1,3,6-trisulfonic acid. Biochem. J. 270, 705–713.

    PubMed  CAS  Google Scholar 

  28. Starr, C. M., Masada, R. I., Hague, C., Skop, E., and Klock, J. C. (1996) Fluorophore-assisted carbohydrate electrophoresis in the separation, analysis, and sequencing of carbohyrates. J. Chromatogr. A 720, 295–321.

    Article  PubMed  CAS  Google Scholar 

  29. Taga, A., Uegaki, K., Yabusakao, Y., Kitano, A., and Honda, S. (1999) Simultaneous determination of the association constant of oligosaccharides to a lectin by capillary electrophoresis. J. Chromatogr. A 837, 221–229.

    Article  CAS  Google Scholar 

  30. Domon, B. and Costello, C.E. (1988) A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates. Glycoconjugate J. 5, 397–409.

    Article  CAS  Google Scholar 

  31. Charlwood, J., Langridge J., and Camilleri, P. (1999) Structural characterisation of N-linked glycan mixtures by precursor ion scanning and tandem mass spectrometric analysis. Rapid Commun. Mass Spectrom. 13, 1522–1530.

    Article  PubMed  CAS  Google Scholar 

  32. Charlwood, J., Birrell, H., Gribble, A., Burdes, V., Tolson, D., and Camilleri, P., (2000) A probe for the versatile analysis and characterization of N-linked oligosaccharides. Analyt. Chem. 72, 1453–1461.

    Article  CAS  Google Scholar 

  33. Kelly, J. F., Locke, S. J., Ramaley, L., and Thibault, P. (1996) Development of electrophoretic conditions for the characterization of protein glycoforms by capillary electrophoresis-electrospray mass spectrometry. J. Chromatogr. A 720, 409–427.

    Article  PubMed  CAS  Google Scholar 

  34. Bateman, K. P., White, R. L., Yaguchi, M., and Thibault, P. (1998) Characterization of protein glycoforms by capillary electrophoresis-nanoelectropsray mass spectrometry. J. Chromatogr. A 794, 327–344.

    Article  CAS  Google Scholar 

  35. Domon, B., Mueller, R. D., and Richter W. J., (1994) Tandem mass spectrometric analysis of fixed-charge derivatized oligosaccharides. Org. Mass Spectrom. 29, 713–719.

    Article  CAS  Google Scholar 

  36. Lindardt R. J. (1994) Capillary electrophoresis of oligosaccharides. Methods Enzymol. 230, 265–288.

    Article  Google Scholar 

  37. Bateman, K. P., Thibault, P., and White, R. L. (1997) Disposable emitters for online capillary zone electrophoresis-electrospray mass spectrometry. Rapid Commun. Mass Spectrom. 11, 307–315.

    Article  CAS  Google Scholar 

  38. Foret, F., Szoko, E., and Karger, B. L. (1992) On-column transient and coupled column isotachophoretic preconcentration of protein samples in capillary zone electrophoresis. J. Chromatogr. A 608, 3–12.

    Article  CAS  Google Scholar 

  39. Locke, S. J. and Thibault, P. (1994) Improvement in detection limits for the determination of paralytic shellfish poisoning toxins in shellfish tissues using capillary electrophoresis/electrospray mass spectrometry and discontinuous buffer systems. Analyt. Chem. 66, 3436–3446.

    Article  CAS  Google Scholar 

  40. Huang, M., Vorkink, W. P., and Lee, M. L. (1992) High efficiency cross-linked polyacrylamide coating for capillary electrophoresis of proteins. J. Microcol. Sep. 4, 233–238.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Kinki University, Higashi-osaka, Japan

    Shigeo Suzuki & Susumu Honda

  2. Institute for Biological Sciences, National Research Council of Canada, Ottawa, ON, Canada

    John F. Kelly & Steven J. Locke

  3. Director of Protein Analysis, Caprion Pharmaceuticals, Montreal, Quebec, Canada

    Pierre Thibault

Authors
  1. Shigeo Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John F. Kelly
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Steven J. Locke
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pierre Thibault
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Susumu Honda
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Caprion Pharmaceuticals, Montreal, Quebec, Canada

    Pierre Thibault

  2. Faculty of Pharmaceutical Sciences, Kinki University, Japan

    Susumu Honda

Rights and permissions

Reprints and permissions

Copyright information

© 2003 Humana Press Inc.

About this protocol

Cite this protocol

Suzuki, S., Kelly, J.F., Locke, S.J., Thibault, P., Honda, S. (2003). Derivatization of Carbohydrates. In: Thibault, P., Honda, S. (eds) Capillary Electrophoresis of Carbohydrates. Methods in Molecular Biology™, vol 213. Humana Press. https://doi.org/10.1385/1-59259-294-5:41

Download citation

  • DOI: https://doi.org/10.1385/1-59259-294-5:41

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-826-4

  • Online ISBN: 978-1-59259-294-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation