Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Amperometric Determination of Urea Using Enzyme-Modified Carbon Paste Electrode

  • Yang, Jae-Kyeong (Laboratory of Forest Chemistry, Faculty of Forest Science, Gyeongsang National University) ;
  • Ha, Kwang-Soo (Department of Chemistry and Research Institute of Natural Science, Gyeongsang National University) ;
  • Baek, Hyun-Sook (Department of Chemistry and Research Institute of Natural Science, Gyeongsang National University) ;
  • Lee, Shim-Sung (Department of Chemistry and Research Institute of Natural Science, Gyeongsang National University) ;
  • Seo, Moo-Lyong (Department of Chemistry and Research Institute of Natural Science, Gyeongsang National University)
  • Published : 2004.10.20
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Abstract

An amperometric biosensor based on carbon paste electrodes (CPEs) for the determination of urea was constructed by enzyme (urease/GL-DH)-modified method. Urea was hydrolyzed to ${NH_4}^+$ by catalyzing urease onto the enzyme-modified electrode surface in sample solution. In the presence of ${\alpha}$-ketoglutarate and reduced nicotinamide adenine dinucleotide(NADH), a liberated ${NH_4}^+$ produce to L-glutamate and $NAD^+$ by Lglutamate dehydrogenase (GL-DH). After the chemical reaction was proceeded, the electrochemical reaction was occurred that an excess of the NADH was oxidized to $NAD^+$. The oxidation current of NADH was monitored at +1.10 volt vs. Ag/AgCl. An optimum conditions of biosensor were investigated: The optimum pH range for catalyzed hydrolysis reaction of urea was pH 7.0-7.4. The linear response range and detection limit were $2.0\;{\times}\;10^{-5}{\sim}2.0\;{\times}\;10^{-4}M\;and\;5.0\;{\times}\;10^{-6}M$, respectively. Another physiological species did not interfere, except L-ascorbic acid.

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