Amino oxidase amperometric biosensor for polyamines |
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| Institution: | 1. Department of Biological Chemistry, Padua University, Via Trieste 75, 35100 Padua Italy;2. Department of Physical Chemistry, Venice University, Venice Italy;1. Food Microbiology, Wageningen University & Research, PO Box 17, 6700 AA, Wageningen, the Netherlands;2. Food Safety Authority of Ireland, The Exchange, Georges Dock, IFSC, Dublin, D01P2V6, Ireland;3. JM Farber Global Food Safety, University of Guelph, Guelph, Ontario, K1A OL2, Canada;4. Food Safety Futures, 6524 BS, Nijmegen, the Netherlands;1. Department of Pharmacology, School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, China;2. Department of Food and Pharmaceutical Engineering, School of Pharmacy, Nanjing Normal University, Nanjing, 210046, China;3. Institute of Plant Resources and Chemistry, Nanjing Research Institute for Comprehensive Utilization of Wild Plants, Nanjing, 210042, China;4. NICE Zhejiang Technology Co. Ltd., Hangzhou, 310051, China;5. Key Laboratory of Green Cleaning Technology & Detergent of Zhejiang Province, Lishui, 323000, China;6. Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, China;1. Department of Analytical Chemistry and Applied Spectroscopy, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin Str., 87-100, Toruń, Poland;2. Department of Organic Chemistry, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin Str., 87-100, Toruń, Poland |
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| Abstract: | An improved amino oxidase enzyme electrode has been constructed and applied to the determination of the amount of polyamines present in real samples. The electrode is based on the amperometric detection of H2O2 produced in the enzymatic oxidation of polyamines by amino oxidase. Amino oxidase from soybean seedlings, characterized by an extremely high activity for cadaverine and putrescine, was used. The enzyme was immobilized in an agarose matrix in the presence of glutaraldehyde and bovine serum albumin on the surface of a Pt electrode. Cadaverine, in concentrations between 0.5 and 500 μM, can be quantitatively determined by use of the amino oxidase electrode, the linear calibration range being 0.5–10 μM. The lower detection limit was 0.2 μM and the response time was 15 to 60 s. Putrescine showed similar behaviour. The maximum current response for cadaverine was 5.1 μA/cm2, with an apparent Michaelis-Menten constant (Km′) of 0.175 mM. The sensor response was stable for more than 32 hours of continuous operation at room temperature and, in the presence of fish or meat homogenates, no change in the signal-to-noise ratio was observed. The long-term stability, pH and temperature response of the biosensor has also been studied. |
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