Highly sensitive flow-biosensor for toxic phenolic compounds using tyrosinase and acridine orange-adsorbed carbon felt |
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| Authors: | Yue WANG Yasushi HASEBE |
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| Institution: | 1. Department of Material Science and Engineering, Saitama Institute of Technology, Fukaya 369-0293, Japan;2. School of Chemical Engineering, University of Science and Technology Liaoning, Anshan 114501, China;3. Department of Life Science and Green Chemistry, Saitama Institute of Technology, Fukaya 369-0293, Japan;1. University of North Texas, Department of Physics and Advanced Materials & Manufacturing Institute, 1155 Union Circle, Denton, TX 76203, USA;2. Military University of Technology, Institute of Optoelectronics, 2 Kaliskiego Str., 00-908 Warsaw, Poland;3. Skolkovo Institute of Science and Technology, 3 Nobel St., Moscow 143026, Russia;1. Department of Analytical Chemistry, Faculty of Chemistry, University of Kashan, Kashan, Iran;2. Department of Chemical Engineering, Laboratory of Nanotechnology, Quchan University of Advanced Technology, Quchan, Iran;1. Department of Life Science and Green Chemistry, Faculty of Engineering, Saitama Institute of Technology, Fukaya, Saitama 369-0293, Japan;2. School of Chemical Engineering, University of Science and Technology Liaoning, Anshan 114501, China |
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| Abstract: | Tyrosinase (TYR, EC 1.14.18.1) was physically adsorbed onto a carbon felt (CF) together with acridine orange (AO). Coadsorption of AO was essential to prevent the denaturation of the TYR at the CF surface. The resulting TYR and AO-coadsorbed CF (TYR/AO-CF) was successfully utilized as a detection unit of novel and highly sensitive amperometric flow-biosensor for toxic chlorophenol compounds. Standard solutions of phenolic compounds (200 μL) were injected, and the cathodic peak currents due to the reduction current of o-quinones produced by the TYR-catalyzed oxidation (phenolase activity) were detected at the applied potential of ?50 mV vs. Ag/AgCl. In this reaction, the electrochemically generated catechol compounds from o-quinones are re-oxidized repeatedly by catecholase activity of the TYR, leading to a sufficient amplified signal. The TYR/AO-CF exhibited much higher selectivity toward p-chlorophenol as compared with other chlorophenol compounds. When 0.1 mol/L phosphate buffer (pH 7.0) was used as a carrier at flow rate of 3.0 mL/min, cathodic peaks for p-chlorophenol was linear in the concentration range between 0.1 and 10 xmol/L (sensitivity: 1.41(mA-L)/mmol) with sampling rate (30 samples/h), and the detection limit ofp-chlorophenol was found to be 2.13 ? 108 mol/L (S/N = 3. The ratio of signal and noise is 3). The TYR/AO-CF kept more than 80% of original activity after the storage in 0.1 mol/L phosphate buffer (pH 7.0) containing 0.2 mmol/L AO at 4°C. |
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