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Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods.
Anal Biochem. 1992 Nov 15; 207(1):176-85.AB

Abstract

Sulfuric acid hydrolysis according to the Saeman procedure, TFA hydrolysis, and methanolysis combined with TFA hydrolysis were compared for the hydrolysis of water-soluble uronic acid-containing polysaccharides originating from fungi, plants, and animals. The constituent sugar residues released were subsequently analyzed by either conventional GLC analysis of alditol acetates or high-performance anion-exchange chromatography with pulsed-amperometric detection. It was shown that TFA hydrolysis alone is not sufficient for complete hydrolysis. Sulfuric acid hydrolysis of these polysaccharides resulted in low recoveries of 6-deoxy-sugar residues. Best results were obtained by methanolysis combined with TFA hydrolysis. Methanolysis with 2 M HCl prior to TFA hydrolysis resulted in complete liberation of monosaccharides from pectic material and from most fungal and animal polysaccharides tested. Any incomplete hydrolysis could be assessed easily by HPAEC, by the detection of characteristic oligomeric products, which is difficult using alternative methods currently in use. Methanolysis followed by TFA hydrolysis of 20 micrograms water-soluble uronic acid containing polysaccharides and subsequent analysis of the liberated sugar residues by HPAEC allowed us to determine the carbohydrate composition of these polysaccharides rapidly and accurately in one assay without the need for derivatization.

Authors+Show Affiliations

De Ruiter GA
Wageningen Agricultural University, Department of Food Science, The Netherlands.
Schols HA
No affiliation info available
Voragen AG
No affiliation info available
Rombouts FM
No affiliation info available

MeSH

AnimalsAnionsCarbohydratesChromatography, High Pressure LiquidChromatography, Ion ExchangeExtracellular SpaceFungiGlucuronatesGlucuronic AcidGlycosaminoglycansHexuronic AcidsHydrolysisIduronic AcidMethanolPectinsPlantsPolysaccharidesSensitivity and SpecificitySolubilityTrifluoroacetic AcidUronic AcidsWater

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

1489092

Citation

De Ruiter, G A., et al. "Carbohydrate Analysis of Water-soluble Uronic Acid-containing Polysaccharides With High-performance Anion-exchange Chromatography Using Methanolysis Combined With TFA Hydrolysis Is Superior to Four Other Methods." Analytical Biochemistry, vol. 207, no. 1, 1992, pp. 176-85.
De Ruiter GA, Schols HA, Voragen AG, et al. Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods. Anal Biochem. 1992;207(1):176-85.
De Ruiter, G. A., Schols, H. A., Voragen, A. G., & Rombouts, F. M. (1992). Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods. Analytical Biochemistry, 207(1), 176-85.
De Ruiter GA, et al. Carbohydrate Analysis of Water-soluble Uronic Acid-containing Polysaccharides With High-performance Anion-exchange Chromatography Using Methanolysis Combined With TFA Hydrolysis Is Superior to Four Other Methods. Anal Biochem. 1992 Nov 15;207(1):176-85. PubMed PMID: 1489092.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods. AU - De Ruiter,G A, AU - Schols,H A, AU - Voragen,A G, AU - Rombouts,F M, PY - 1992/11/15/pubmed PY - 1992/11/15/medline PY - 1992/11/15/entrez SP - 176 EP - 85 JF - Analytical biochemistry JO - Anal Biochem VL - 207 IS - 1 N2 - Sulfuric acid hydrolysis according to the Saeman procedure, TFA hydrolysis, and methanolysis combined with TFA hydrolysis were compared for the hydrolysis of water-soluble uronic acid-containing polysaccharides originating from fungi, plants, and animals. The constituent sugar residues released were subsequently analyzed by either conventional GLC analysis of alditol acetates or high-performance anion-exchange chromatography with pulsed-amperometric detection. It was shown that TFA hydrolysis alone is not sufficient for complete hydrolysis. Sulfuric acid hydrolysis of these polysaccharides resulted in low recoveries of 6-deoxy-sugar residues. Best results were obtained by methanolysis combined with TFA hydrolysis. Methanolysis with 2 M HCl prior to TFA hydrolysis resulted in complete liberation of monosaccharides from pectic material and from most fungal and animal polysaccharides tested. Any incomplete hydrolysis could be assessed easily by HPAEC, by the detection of characteristic oligomeric products, which is difficult using alternative methods currently in use. Methanolysis followed by TFA hydrolysis of 20 micrograms water-soluble uronic acid containing polysaccharides and subsequent analysis of the liberated sugar residues by HPAEC allowed us to determine the carbohydrate composition of these polysaccharides rapidly and accurately in one assay without the need for derivatization. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/1489092/Carbohydrate_analysis_of_water_soluble_uronic_acid_containing_polysaccharides_with_high_performance_anion_exchange_chromatography_using_methanolysis_combined_with_TFA_hydrolysis_is_superior_to_four_other_methods_ DB - PRIME DP - Unbound Medicine ER -