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Amperometric biosensor with HRP immobilized on a sandwiched nano-Au / polymerized m-phenylenediamine film and ferrocene mediator

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Abstract.

An amperometric biosensor has been developed for the determination of H2O2 in plant samples. Horseradish peroxidase (HRP) is immobilized on a sandwiched nano-Au particle / m-phenylenediamine polymer film by glutaraldehyde cross-linking. The film is formulated on the carbon paste electrode (CPE) blended with ferrocene as an electron transfer mediator. On the low concentration range, the current response is related to the H2O2 concentration linearly from 0 to 8×10-6 M with a detection limit of 1.3×10-7 M. On a wider concentration range of 8×10-6 to 1.4×10-4 M, the reciprocal of current response is linearly related to the reciprocal of H2O2 concentration. The apparent Michaelis-Menten constant (Km app) was calculated to be 0.0334 mM. The sensor has been tested by determining H2O2 concentration in plant leaf samples.

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Acknowledgements.

This work was supported by the National Natural Science Foundation of China (Grants Nos. 29975006, 20075006, 70171055, 50179011), the Foundation of The National 863 High Technologies Research of China (No 2001AA644020), the Foundation for Ph.D Thesis Research (No. 20010532008) and the Foundation of Hunan province (Grants Nos. 01JJY1003, 01JJY2099).

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Authors and Affiliations

  1. State Key Laboratory of Chemo/Biosensing and Chemometrics, Chemistry and Chemical Engineering College, Department of Environmental Science and Engineering, Hunan University, 410082, Changsha , P.R. China

    Jin Li, Xuan-Ming Liu, Guang-Ming Zeng, Guo-He Huang, Guo-Li Shen & Ru-Qin Yu

  2. Key Laboratory of Phytohormones of Hunan Province, Hunan Agricultural University, 410028, Changsha, P.R. China

    Lang-Tao Xiao

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  1. Jin Li
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  2. Lang-Tao Xiao
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  3. Xuan-Ming Liu
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Correspondence to Guo-Li Shen.

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Li, J., Xiao, LT., Liu, XM. et al. Amperometric biosensor with HRP immobilized on a sandwiched nano-Au / polymerized m-phenylenediamine film and ferrocene mediator. Anal Bioanal Chem 376, 902–907 (2003). https://doi.org/10.1007/s00216-003-1989-8

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  • DOI: https://doi.org/10.1007/s00216-003-1989-8

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