Abstract
This detection method makes use of sequential immobilized enzyme reactors (IMER) to first hydrolyze β-D-glucosides to β-D-glucose (using β-glucosidase) and then to produce hydrogen peroxide from the β-D-glucose (using glucose oxidase); the hydrogen peroxide is then detected with luminol chemiluminescence. This method has been used for the determination of individual β-D-glucosides (phenyl, p-nitrophenyl, and salicin) via flow-injection analysis and has been extended to the determination of a mixture of glucosides following their separation via reversed-phase high-performance liquid chromatography. The use of up to 30% acetonitrile in the buffered mobile phase had very little effect on the efficiency of the β-glucosidase and glucose oxidase enzyme reactors; overall the use of acetonitrile did not affect the linear working range or detection limits for the glucosides. Chromatographic detection limits are approximately 0.1 μ M (2 pmol) with a linear working range of more than three decades.
Published Version
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