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Biomimetic biosensor based on lipidic layers containing tyrosinase and lutetium bisphthalocyanine for the detection of antioxidants

Year: 2011

Journal: Biosensors and Bioelectronics, 2011, 26 (5), 2513-2519, 20131009

Authors: C. Apetrei, P. Alessio, C.J.L. Constantino, J.A. de Saja, M.L. Rodriguez-Mendez, F.J. Pavinatto, E. Giuliani Ramos Fernandes, V. Zucolotto, O.N. Oliveira Jr.

Organizations: Department of Chemistry, Faculty of Sciences, “Dunărea de Jos” University of Galati, 800008 Galati, Romania; Faculdade de Ciências e Tecnologia, UNESP Univ Estadual Paulista, 19060-900 Presidente Prudente, SP, Brazil; Condensed Matter Physics Department, Faculty of Science, University of Valladolid, 47011 Valladolid, Spain; Department of Inorganic Chemistry, E. T. S. Ingenieros Industriales, University of Valladolid, Paseo del Cauce, 59, 47011 Valladolid, Spain; Instituto de Física de São Carlos, Universidade de São Paulo, CP 369, 13560-970 São Carlos, SP, Brazil

This paper describes the preparation of a biomimetic Langmuir–Blodgett film of tyrosinase incorporated in a lipidic layer and the use of lutetium bisphthalocyanine as an electron mediator for the voltammetric detection of phenol derivatives, which include one monophenol (vanillic acid), two diphenols (catechol and caffeic acid) and two triphenols (gallic acid and pyrogallol). The first redox process of the voltammetric responses is associated with the reduction of the enzymatically formed o-quinone and is favoured by the lutetium bisphthalocyanine because significant signal amplification is observed, while the second is associated with the electrochemical oxidation of the antioxidant and occurs at lower potentials in the presence of an electron mediator. The biosensor shows low detection limit (1.98 × 10−6–27.49 × 10−6 M), good reproducibility, and high affinity to antioxidants (KM in the range of 62.31–144.87 μM). The excellent functionality of the enzyme obtained using a biomimetic immobilisation method, the selectivity afforded by enzyme catalysis, the signal enhancement caused by the lutetium bisphthalocyanine mediator and the increased selectivity of the curves due to the occurrence of two redox processes make these sensors exceptionally suitable for the detection of phenolic compounds.