Chemiluminescence flow biosensor for hydrogen peroxide with immobilized reagents

A novel chemiluminescence (CL) flow biosensor for hydrogen peroxide is described. It is prepared by immobilizing HRP with the sol–gel method on the flow CL cell, and by electrostatically immobilizing luminol on a strongly basic anion-exchanger resin. Hydrogen peroxide is sensed by the CL reaction with HRP sensitive membrane and luminol bleeding from the ion-exchanger column with immobilized luminol by hydrolysis. The calibration graph is linear in the range of 1×10−4–8×10−7 mol l−1, and the detection limit is 2.8×10−7 mol l−1 hydrogen peroxide. A complete analysis could be performed in 1 min with a relative standard deviation of 2.1% for 1×10−6 mol l−1 H2O2 (n=11). The biosensor has been applied successfully to the determination of hydrogen peroxide in rainwater.