Amperometric biosensor based on reductive H2O2 detection using pentacyanoferrate-bound polymer for creatinine determination |
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Authors: | Chi-Hua Nieh Seiya Tsujimura Osamu Shirai Kenji Kano |
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Affiliation: | 1. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University Sakyo, Kyoto 606-8502, Japan;2. Division of Materials Science, Faculty of Pure and Applied Sciences, University of Tsukuba 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan |
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Abstract: | Pentacyanoferrate-bound poly(1-vinylimidazole) (PVI[Fe(CN)5]) was selected as a mediator for amperometric creatinine determination based on the reductive H2O2 detection. Creatinine amidohydrolase (CNH), creatine amidohydrolase (CRH), sarcosine oxidase (SOD), peroxidase (POD), and PVI[Fe(CN)5] were crosslinked with poly(ethylene glycol) diglycidyl ether (PEGDGE) on a glassy carbon (GC) electrode for a creatinine biosensor fabrication. Reduction current was monitored at −0.1 V in the presence of creatinine and O2. It is revealed that PVI[Fe(CN)5] is suitable as a mediator for a bioelectrocatalytic reaction of POD, since PVI[Fe(CN)5] neither reacts with reactants nor works as an electron acceptor of SOD. The amounts of PVI[Fe(CN)5], PEGDGE, and enzymes were optimized toward creatinine detection. Nafion as a protecting film successfully prevented the enzyme layer from interferences. The detection limit and linear range in creatinine determination were 12 μM and 12–500 μM (R2 = 0.993), respectively, and the sensitivity was 11 mA cm−2 M−1, which is applicable for urine creatinine tests. The results of the creatinine determination for four urine samples measured with this proposed method were compared with Jaffe method, and a good correlation was obtained between the results. |
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Keywords: | Pentacyanoferrate-bound poly(1-vinylimidazole) Creatinine amidohydrolase Reductive H2O2 detection Peroxidase Nafion Urine creatinine test |
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