Amine oxidase amperometric biosensor coupled to liquid chromatography for biogenic amines determination |
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Authors: | Laura Mureşan Ruben Ronda Valera Ivo Frébort Ionel Catalin Popescu Elisabeth Csöregi Mihaela Nistor |
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Institution: | 1. Department of Physical Chemistry, “Babe?-Bolyai” University, Arany Janos 11, RO-400028, Cluj-Napoca, Romania 2. Department of Analytical Chemistry, Lund University, P.O. Box 124, SE-22100, Lund, Sweden 3. Department of Biochemistry, Palacky University, ?lechtitel? 11, CZ-78371, Olomouc, Czech Republic
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Abstract: | A selective and sensitive method is presented for biogenic amines (BA) determination. The novelty consists in coupling a highly selective electrochemical biosensor to a weak acid cation-exchange column for online detection of amines. A bienzyme design, based on a recently isolated amine oxidase from grass pea and commercial horseradish peroxidase, was used for the biosensor construction. The enzymes were co-immobilized on the surface of a graphite electrode together with the electrochemical mediator (Os-redox polymer). The electrochemical detection was performed at a low applied potential (?50 mV vs. Ag/AgCl, KCl0.1 M), where biases from interferences are minimal. The separation and determination of six BA, with relevance in food analysis (tyramine, putrescine, cadaverine, histamine, agmatine and spermidine), were investigated. Irrespective of the BA nature, the amine oxidase-based biosensor showed a linear response up to 5 mM, and its sensitivity decreases in the following order: cadaverine, putrescine, spermidine, agmatine, histamine and tyramine. The approach was used to estimate the BA content in fish samples, after their extraction with methanesulfonic acid. |
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