Quantitative anhydrous mercaptolysis of algal galactans followed by HPLC of component sugars

This paper describes a new HPLC method for the sugar analysis of algal galactans containing 3,6-anhydrogalactose, which is readily destroyed during usual acid hydrolysis and methanolysis. By anhydrous mercaptolysis of galactans at 60°C for 6 h with the newly developed solvent system, 0.5 N HCl/[etha...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1999, Vol.125 (1), p.160-165
Main Authors: Hama, Y, Nakagawa, H, Mochizuki, K, Sumi, T, Hatate, H
Format: Article
Language:English
Subjects:
Absorption
algae
algal galactans
anhydrogalactose
anhydrous mercaptolysis
Biochemistry - methods
Brackish
Carbohydrates - analysis
chemical composition
Chromatography, High Pressure Liquid - methods
derivatives
dithioacetal
Eukaryota - chemistry
Freshwater
galactans
Galactans - analysis
Galactans - chemistry
Galactose - analogs & derivatives
Galactose - analysis
Galactose - chemistry
high performance liquid chromatography
HPLC
Marine
Rhodophycota
Spectrophotometry, Ultraviolet
sugar diethyl dithioacetals
sugars
Sulfhydryl Compounds - chemistry
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Summary:This paper describes a new HPLC method for the sugar analysis of algal galactans containing 3,6-anhydrogalactose, which is readily destroyed during usual acid hydrolysis and methanolysis. By anhydrous mercaptolysis of galactans at 60°C for 6 h with the newly developed solvent system, 0.5 N HCl/[ethanethiol:methanol (2:1, v/v)], component sugars, including 3,6-arihydrogalactose, were liberated quantitatively as their diethyl dithio-acetals. The resultant sugar diethyl dithioacetals were found to have strong UV-absorption (absorption maximum of 3,6-anhydrogalactose diethyl dithioacetal in water, 191–192 nm; molar extinction coefficient, 4,400). The sugar diethyl dithioacetals were then resolved simultaneously within 15 min by reversed phase HPLC with 30% acetonitrile as the eluent and detected by UV-absorption at 215 nm. Amounts of sugar diethyl dithioacetals less than 50 pmol can be determined without further derivatization.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a022253