CHARACTERIZATION OF AN IMMOBILIZED HEXOSE OXIDASE REACTOR FOR MONOSACCHARIDE AND OLIGOSACCHARIDE DETERMINATION BY LIQUID-CHROMATOGRAPHY

The enzyme hexose oxidase was extracted from the red sea-weed Chondrus crispus and immobilized in an enzyme reactor. The characteristics of this reactor were studied for the post-column liquid chromatographic detection of carbohydrates (separation on a cation exchange column). The reactor converted the eluting carbohydrates to hydrogen peroxide, which was detected amperometrically. Several saccharides could be determined at low mu g levels. Conversion efficiencies for 12 mono- and oligosaccharides were measured at different flow-rates. The flow-rate versus conversion efficiency behaviour of the reactor and its selectivity is compared to the behaviour of a glucose oxidase reactor with very high enzyme loading in this respect. The differences between both reactors are explained theoretically. The practical use of both reactors in chromatographic analysis of carbohydrates is discussed and demonstrated.