Composite multienzyme amperometric biosensors for an improved detection of phenolic compounds

A biosensor design, in which glucose oxidase and peroxidase are coimmobilized by simple physical inclusion into the bulk of graphite-Teflon pellets, is reported for the detection of phenolic compounds. This design allows the in situ generation of the H2O2 needed for the enzyme reaction with the phenolic compounds, which avoids several problems detected in the performance of single peroxidase biosensors as a consequence of the presence of a high H2O2 concentration. So, a much lower surface fouling was found at the GOD-HRP biosensor in comparison with a graphite-Teflon-HRP electrode, suggesting that the controlled generation of H2O2 makes more difficult the formation of polymers from the enzyme reaction products. The construction of trienzyme biosensors, in which GOD, HRP and tyrosinase were coimmobilized into the graphite-Teflon matrix is also reported, and their performance was compared with that of GOD-HRP bienzyme electrodes. The practical applicability of the composite multienzyme amperometric biosensors was evaluated by the estimation of the phenolic compounds content in waste waters from a refinery, and the results were compared with those obtained by using a colorimetric official method based on the reaction with 4-aminoantipyrine.