Development of a bienzyme system based on sugar-lectin biospecific interactions for amperometric determination of phenols and aromatic amines

Bienzyme electrode with horseradish peroxidase (HRP) and glucose oxidase (GOD) multilayers was constructed based on sugar-lectin biospecific interactions for amperometric determination of phenolic compounds and aromatic amines. Atomic force microscopy (AFM) was applied to monitor the uniform layer-by-layer assembly of concanavalin A (Con A) and HRP or GOD on polyelectrolyte precursor film-modified Au electrode. Substituted phenolic compounds and aromatic amines could be determined with in situ generation of H2O2 by GOD-catalyzed oxidation of glucose. The parameters of the biosensor including the number of assembled HRP and GOD layer, and the concentrations of glucose were optimized. The linear range for the determination of catechol and p-phenyldiamine was 6.0-60.0 mu mol L-1 and 7.6-68.4 mu mol L-1 with detection limit of 0.9 mu mol L-1 and 0.4 mu mol L-1, respectively. The biosensor possessed high sensitivity and fast response for phenolic compounds and 95% of the maximum response could be reached in about 3 s. Glucose, ascorbic acid, tartaric acid, citric acid and starch exhibited no interference for the detection. The biosensor presented high stability due to the design for in situ generation of H2O2 with bienzyme system. (c) 2007 Elsevier B.V. All rights reserved.