Immobilization of glucosidase onto silica-based, amino functionalized beads for enzymatic hydrolysis of urinary phenol prior to liquid chromatographic analysis
- Analytica Chimica Acta, 292(1-2), p.23-30, 1994 .
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