A new approach for nitrite determination based on a HRP/catalase biosensor

Abstract In this paper, a new concept of enzyme inhibition-based biosensor involving a two enzyme system was developed. The latter displays a signal increase instead of a decrease in the presence of an inhibitor. HRP and catalase were thus separately entrapped into Layered Double Hydroxides (LDH), an anionic clay, as a host matrix. The inner layer was constituted of HRP electrically wired by [Zn 2 CrABTS] LDH and the outer layer contained catalase immobilized in [Zn 3 AlCl] LDH. Both enzymes catalyzed the decomposition of H 2 O 2 , HRP its reduction and catalase its breakdown into oxygen and water. Nitrite was selected as a specific inhibitor of catalase. In the presence of H 2 O 2 , the nitrite addition blocked the H 2 O 2 consumption by catalase, inducing thus an increase in the amperometric signal of the H 2 O 2 reduction at 0 V by the wired HRP. The optimum configuration of the bi-enzyme biosensor displayed in aerated aqueous solutions, a nitrite sensitivity of 102 μA M − 1 · cm − 2 with a fast response time, the detective limit being 4 μM.

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