Immobilization of glucosidase onto silica-based, amino functionalized beads for enzymatic hydrolysis of urinary phenol prior to liquid chromatographic analysis
✍ Scribed by Jen-Fon Jen; Jen-Haw Zen; Fu-Chou Cheng; Goang-Yean Yang
- Publisher
- Elsevier Science
- Year
- 1994
- Tongue
- English
- Weight
- 655 KB
- Volume
- 292
- Category
- Article
- ISSN
- 0003-2670
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✦ Synopsis
An amino-functionalized silica commercial liquid chromatography packing material was used instead of controlled-pore glass as the support to react with glutaraldehyde and then to directly immobilize the /3-glucosidase. After enzymatic hydrolysis, the urinary phenol was separated by reversed-phase liquid chromatography and monitored by an electrochemical detector at 0.85 V (vs. Ag/AgCl). Factors that affect the immobilization and hydrolysis were investigated. The proposed method provides a simpler procedure with a less expensive material for glucosidase immobilization, and the electrochemical detection avoids the interference from other urinary species and hence simplifies the chromatogram obtained. The detection limit was 0.2 ng with a 28 ~1 injection. The relative standard deviations for 1.2 ng and 2.0 ng levels of phenols were 1.16% and 3.38%, respectively.