Separation of ascorbic acid, dehydroascorbic acid, diketogulonic acid and glucose by isocratic elution from a column of a hydrophilic gel.
High-performance liquid chromatography on an Asahipak GS-320 hydrophilic gel column with tartrate buffer (0.015 M, pH 3.0) containing 2 mM ethylenediaminetetraacetate and 0.05% beta-thiodiglycol as the eluent allowed the separation of glucose, diketogulonic acid, dehydroascorbic acid and ascorbic acid within 30 min. Fluorimetric monitoring of these compounds in the eluate with benzamidine at alkaline pH and at 90 degrees C in the presence of potassium sulphite allowed the determination of nanogram amounts of ascorbic acid, dehydroascorbic acid and diketogulonic acid. This method was applied to the determination of ascorbic acid in fruit juice.
D015083:2,3-Diketogulonic Acid / Q000302:isolation & purification*; D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000302:isolation & purification*; D002850:Chromatography, Gel; D003683:Dehydroascorbic Acid / Q000302:isolation & purification*; D005638:Fruit / Q000032:analysis; D005947:Glucose / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence; D013400:Sugar Acids / Q000302:isolation & purification*
D013400:Sugar Acids; D015083:2,3-Diketogulonic Acid; D005947:Glucose; D001205:Ascorbic Acid; D003683:Dehydroascorbic Acid