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Journal of Chromatography A
Volume 1181, Issues 1-2, 15 February 2008, Pages 45-50
 
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doi:10.1016/j.chroma.2007.12.027    How to Cite or Link Using DOI (Opens New Window)
Copyright © 2007 Elsevier B.V. All rights reserved.

Determination of galactose and mannose residues in natural galactomannans using a fast and efficient high-performance liquid chromatography/UV detection

N. Tapiea, C. MalhiacCorresponding Author Contact Information, a, E-mail The Corresponding Author, N. Huchera and M. Grisela

aUnité de Recherche en Chimie Organique et Macromoléculaire, URCOM, EA 3221, Université du Havre, 25 rue P. Lebon, B.P. 540, F-76058 Le Havre cedex, France

Received 31 July 2007; 
revised 5 December 2007; 
accepted 12 December 2007. 
Available online 16 December 2007.

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Abstract

The present work describes the validation of an easy, fast and efficient precolumn derivatization method for the quantification of oligosides, mannose and galactose obtained by degradation of galactomannans. This work combines an acid hydrolysis and an enzymatic degradation of natural galactomannans with the quantification of released residues by reversed-phase HPLC–UV, the most usual HPLC system in laboratories. In case of enzymatic degradation, mannotetraose has been detected and quantified for the first time, and an application to the evaluation of the galactosyl distribution in galactomannans is proposed. After an acidic hydrolysis, this method also allowed to obtain the mannose/galactose (M/G) ratio.

Keywords: Galactomannan; Enzymatic degradation; β-Mannanase; Mannose/galactose ratio; Reversed-phase HPLC–UV detection

Article Outline

1. Introduction
2. Experimental
2.1. Reagents
2.2. Enzymatic degradation of galactomannans
2.3. Precolumn derivatization
2.4. Chromatographic conditions
2.5. Stock solutions and calibration standard preparation
3. Results and discussion
3.1. Performance of the procedure
3.1.1. Validation of the method
3.1.2. Quantification procedure
3.1.3. Control cards: application to a guar gum and a tara gum
3.2. Application to the evaluation of the galactosyl distribution in galactomannans
4. Conclusion
References





 
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