A procedure for the determination of galacturonic acid and the main neutral sugars in pectins involves enzymic hydrolysis followed by methanolysis and h.p.1.c. The usefulness of this method was demonstrated by comparison of the results obtained by (I) methanolysis in methanolic M HCI without enzymic
Assessment of methanolysis for the determination of composite sugars of gelling carrageenans and agarose by HPLC
✍ Scribed by Bernard Quemener; Marc Lahaye; François Metro
- Publisher
- Elsevier Science
- Year
- 1995
- Tongue
- English
- Weight
- 626 KB
- Volume
- 266
- Category
- Article
- ISSN
- 0008-6215
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✦ Synopsis
The characterization of the main composite sugars of commercial gelling red algae galactans (agarose, iota and kappa carrageenans) by methanolysis and separation of the methyl glycosides produced by high performance liquid chromatography is described. The methanolysis (methanolic hydrocMoric acid strength, temperature, and reaction time) was optimized in order to release monosaccharides in near quantitative yield. The results were compared to those obtained by (1) gas chromatography of the alditol acetates of the neutral sugars released by acid hydrolysis and (2) specific colorimetric determination of the acid-labile 3,6-anhydrogalactosyl residue. Conditions such as methanolic 0.125 M HCI for 1 h at 85°C were sufficient to release all of the galactosidic and 3,6-anhydrogalactosidic bonds for iota carrageenan without apparent degradation of the anhydrogalactosyl unit. However, with the same conditions, the yields of 3,6-anhydrogalactosyl residues were 80 and 70% for kappa carrageenan and agarose, respectively. These yields were not improved by stronger conditions. At the opposite extreme, under very mild methanolysis conditions such as methanolic 0.01 M HCI at 100°C for 1 h, agars and gelling carrageenans were well differentiated by the respective determination of agarobiose-and carrabiose-dimethyl acetal which are well-separated on octadecyl reversed phase HPLC columns with water as eluent.
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