Abstract
Flow-amperometric microdetermination of heavy metal ions based on an apoenzyme reactivation method is proposed. A hybrid type of enxyme membrane was prepared by co-immobilizing alkaline phosphatase (zinc enzyme) and ascorbate oxidase (copper enzyme) onto a porous polymer membrane and thereby used as the recognition element for zinc(II) and copper(II) ions. The biosensing system was assembled with the hybrid-enzyme membrane attached to a sensing part of a flow-through oxygen electrode for monitoring the enzymic activity. Zinc(II) ions in 2 to 200 μM levels and also copper(II) ions in 2 to 100 μM levels were amperometrically determined through the activation of each immobilized metal-free enzyme (apoenzyme). The apoenzyme membrane was regenerated by pumping chelating agents to remove each of the heavy metal ions from the catalytic site of alkaline phosphatase or ascorbate oxidase.
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