Sugar determination in ulvans by a chemical-enzymatic method coupled to high performance anion exchange chromatography |
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Authors: | B Quemener M Lahaye C Bobin-Dubigeon |
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Institution: | (1) Laboratoire de Biochimie et Technologie des Glucides, Institut National de la Recherche Agronomique, B.P. 1627, 44316 Nantes Cedex 03, France |
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Abstract: | The sugar determination of ulvans, the water-soluble polysaccharides from Ulva sp. and Enteromorpha sp., was optimized by
combining partial acid prehydrolysis (2 mol L-1 trifluoroacetic acid, 120°C) with enzymic hydrolysis (with β-D-glucuronidase).
The different constitutive sugars (rhamnose, galactose, glucose, xylose, glucuronic acid), released after hydrolysis, were
separated by high performance anion-exchange chromatography and determined by pulsed amperometric detection. The ulvanobiouronic
acid, β-D-GlcA-(1,4)-L-Rha, which is the main constituent of ulvans was always present after 3 h of trifluoroacetic acid hydrolysis
(approx. 2% D.M.) when acid hydrolysis was performed alone but the xylose amount fell to 75% of its maximum value at this
time. The optimal duration of 2 mol L−1 trifluoroacetic acid hydrolysis of ulvans (i.e. without any degradation of xylose, rhamnose and glucuronic acid) was 45 min.
Additionnal treatment of the partial acid hydrolysate by purified β-D-glucuronidase allowed the hydrolysis of the residual
ulvanobiouronic acid in rhamnose and glucuronic acid. High performance anion exchange chromatography coupled to this chemical-enzymic
hydrolysis revealed to be a high resolution chromatographic technique for monitoring the hydrolysis of the aldobiouronic acid
by β-D-glucuronidase. This method allowed the simultaneous quantitative determination of neutral and acidic sugars and revealed
the presence of iduronic acid inulvans.
This revised version was published online in September 2006 with corrections to the Cover Date. |
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Keywords: | aldobiouronic acid Enteromorpha β glucuronidase HPAEC-PAD iduronic acid Ulva Ulvan |
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