Abstract
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 degreesC for 6 h with the newly developed solvent system, 0.5 N HCl/[ethanethiol:methanol (2:1, v/v)], component sugars, including 3,6-anhydrogalactose, were liberated quantitatively as their diethyl dithioacetals. 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.
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