CHARACTERIZATION OF CELLULASE-BASED ENZYME REACTORS FOR THE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF BETA-D-GLUCAN OLIGOSACCHARIDES

Post-column enzyme reactors were used in series with an electrochemical detector for the selective high-performance liquid chromatographic (HPLC) determination of beta-D-glucan oligo- and polysaccharides (degree of polymerization up to 30). Immobilized cellulase converted the eluting oligomers to beta-D-glucose, which was oxidized by immobilized glucose oxidase. The production of hydrogen peroxide was measured with an electrochemical detector. The functioning of this system was verified for a whole range of glucosaccharides varying in both structure (positional isomers) and degree of polymerization. Fractional conversions and molar response factors were determined for all the compounds under study. Rate constants are discussed for the reactor system used applying a first-order kinetics model. Efficient HPLC separations were obtained for these oligo- and polysaccharides on a reversed-phase column using gradient elution. Detection limits were of the order of a few nanograms. The reactors were stable for several months.