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Sugar determination in ulvans by a chemical-enzymatic method coupled to high performance anion exchange chromatography

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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.

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Authors and Affiliations

  1. Laboratoire de Biochimie et Technologie des Glucides, Institut National de la Recherche Agronomique, B.P. 1627, 44316, Nantes Cedex 03, France

    B. Quemener, M. Lahaye & C. Bobin-Dubigeon

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  1. B. Quemener
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  2. M. Lahaye
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  3. C. Bobin-Dubigeon
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Correspondence to M. Lahaye.

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Quemener, B., Lahaye, M. & Bobin-Dubigeon, C. Sugar determination in ulvans by a chemical-enzymatic method coupled to high performance anion exchange chromatography. Journal of Applied Phycology 9, 179–188 (1997). https://doi.org/10.1023/A:1007971023478

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