Amperometric sensors for peroxide, choline, and acetylcholine based on electron transfer between horseradish peroxidase and a redox polymer.

Amperometric sensors have been developed for hydrogen peroxide, choline, and acetylcholine by immobilization of horseradish peroxidase, (HRP), choline oxidase, and acetylcholinesterase in a cross-linked redox polymer deposited on glassy carbon electrodes. Peroxide sensors, prepared by immobilization of HRP alone, gave detection limits of 10 nM and a linear response up to ca. 1 mM. Coimmobilization of HRP and glucose oxidase was used to establish the feasibility of highly efficient bienzyme sensors at low substrate levels. Replacing glucose oxidase with choline oxidase produced sensors with submicromolar detection limits and a linear response up to 0.8 mM. Addition of acetylcholinesterase to the sensors generated a relatively small response to acetylcholine that demonstrates the feasibility of trienzyme sensors. At low substrate concentrations, no loss in sensitivity during a 1-day experiment was observed. The response times of these sensors are all less than 30 s with 2-s response times achieved in some cases.