Abstract
Immobilized sorbitol dehydrogenase was used as a reactor in a column liquid chromatographic system for the selective detection of sucrose and fructose. Sorbitol dehydrogenase was immobilized on controlled-pore glass. The sugars were separated on a cation-exchange resin column with a mobile phase containing 0.1 M phosphate buffer (pH 7.0) and 100 μM NADH. The disappearance of NADH was detected spectrofluorimetrically. For sucrose the linear working range was 50–500 μM, with a detection limit of 10 μM (170 ng in a 50-μl injection); for fructose the range was 25–250 μM, with a detection limit of 5 μM (45 ng in a 50μl injection).
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