Abstract
A flow-through sensor based on transient immobilization of NADH, the product of the reaction between glycerol and NAD+ catalysed by glycerol dehydrogenase, is proposed. The biocatalyst is immobilized on controlled-pore glass, where the enzymic reaction takes place, the product of the reaction being transiently retained and spectrofluorimetrically monitored on passage through a flow cell packed with a solid support. The proposed method affords a linear range between 0.3 and 5.0 µg ml–1 of glycerol, and a detection limit of 0.1 µg ml–1, with a relative standard deviation of less than 2%. The performance of the sensor was checked by applying it to the determination of glycerol in wine. The results obtained agree well with those provided by a conventional method based on HPLC.
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