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Scientific article
English

Determination of sugars by liquid chromatography with post-column catalytic derivatization and fluorescence detection

Contributors

Coquet, A.; Haerdi, Werner; Degli Agosti, Robert; Veuthey, Jean-Luc

Published in

Chromatographia, vol. 38, no. 1-2, p. 12-16

Publication date

1994

Abstract

A method for the determination of reducing sugars such as fructose and glucose and nonreducing sugar such as sucrose by high performance liquid chromatography followed by an acidic hydrolysis and a derivatization with benzamidine has been developed. After separation of sugars on a gel column packed with a polymer-based cation exchange material (Sugar-Pak I, Waters-Millipore), the sucrose is first hydrolysed in a solid phase reactor to convert it into reducing subunits. A post-column fluorigenic reaction with benzamidine under alkaline condition allows the selective determination of both natural and converted reducing carbohydrates. This procedure has proven to be selective (fluorigenic detection) and highly sensitive (allowing detection as little as picomoles amounts), reproducible and linear over a broad range of concentrations: 5×10–4 to 1.0×10–2 M. The applicability of this method to natural matrices such as plant extracts and beverages is also described. The sugar content of a barley extract has been determined and compared with a specific enzymatic test. The determined sugar content of natural and commercial lemon juices as well as of Cola beverages has been compared with those found by the conventional LC refractive index analytical procedure. In all cases, the results were comparable and were within the experimental errors of the methods.

Keywords

  • Column liquid chromatography
  • Carbohydrates
  • Post-column fluorescence derivatization
  • Post column catalytic reaction

UNIGE affiliation entities

Citation (ISO format)

COQUET, A. et al. Determination of sugars by liquid chromatography with post-column catalytic derivatization and fluorescence detection. In: Chromatographia, 1994, vol. 38, n° 1-2, p. 12–16. doi: 10.1007/BF02275720

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Article (Published version)

Identifiers

Additional URL for this publication

http://www.springerlink.com/index/10.1007/BF02275720

Journal ISSN

0009-5893
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Technical informations

Creation

22/06/2010 15:05:00

First validation

22/06/2010 15:05:00

Update

26/03/2026 10:48:30

Status update

26/03/2026 10:48:30

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