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Analysis of mono-, di- and oligosaccharides by CE using a two-stage derivatization method and LIF detection

  1. Hua-Ming Tseng1,
  2. Stefano Gattolin2,
  3. Jeremy Pritchard2,
  4. H. John Newbury2,
  5. David A. Barrett1,*

Article first published online: 24 MAR 2009

DOI: 10.1002/elps.200800517

Issue

ELECTROPHORESIS

ELECTROPHORESIS

Special Issue: EMPHASIS ON FUNDAMENTALS AND METHODOLOGIES

Volume 30, Issue 8, pages 1399–1405, April 2009

Additional Information

How to Cite

Tseng, H.-M., Gattolin, S., Pritchard, J., Newbury, H. J. and Barrett, D. A. (2009), Analysis of mono-, di- and oligosaccharides by CE using a two-stage derivatization method and LIF detection. ELECTROPHORESIS, 30: 1399–1405. doi: 10.1002/elps.200800517

Author Information

  1. 1

    Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, UK

  2. 2

    School of Biosciences, University of Birmingham, Edgbaston, Birmingham, UK

*Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, UK Fax: +44-115-951-5102

Publication History

  1. Issue published online: 20 APR 2009
  2. Article first published online: 24 MAR 2009
  3. Manuscript Accepted: 30 OCT 2008
  4. Manuscript Revised: 24 OCT 2008
  5. Manuscript Received: 12 AUG 2008

Funded by

  • UK Biotechnology
  • Biological Sciences Research Council (BBSRC)

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Keywords:

  • Carbohydrates;
  • CE;
  • Honeydew;
  • LIF;
  • Phloem sap

Abstract

A sensitive CE with LIF method has been developed for quantitative analysis of small carbohydrates. In this work, 17 carbohydrates including mono-, di- and oligosaccharides were simultaneously derivatized with 4-fluoro-7-nitrobenzofurazane (NBD-F) via a two-step reaction involving reductive amination with ammonia followed by condensation with NBD-F. Under the optimized derivatization conditions all carbohydrates were successfully derivatized within 2.5 h and separated within 15 min using borate buffer (90 mmol/L, pH 9.2). For sugar standards LODs were in the range of 49.7 to 243.6 nmol/L. Migration time and peak area reproducibility were better than RSD 0.1 and 3%, respectively. The method was applied to measure sugars in nanoliter volume samples of phloem sap obtained by stylectomy from wheat and to honeydew samples obtained from aphids feeding from wheat and willow.

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