High-performance liquid chromatographic determination of glucosides (glucose conjugates) with post-column reaction detection combining immobilized enzyme reactors and luminol chemiluminescence

✍ Scribed by Philip J. Koerner Jr.; Timothy A. Nieman

• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 900 KB
• Volume: 449
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(00)94381-4

✦ Synopsis

This detection method makes use of sequential immobilized enzyme reactors (IMER) to first hydrolyze D-D-glucosides to /&D-glucose (using fi-glucosidase) and then to produce hydrogen peroxide from the P-o-glucose (using glucose oxidase); the hydrogen peroxide is then detected with luminol chemiluminescence. This method has been used for the determination of individual fl-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 efIiciency of the fi-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 PM (2 pmol) with a linear working range of more than three decades.