Horseradish Peroxidase Biosensor to Detect Zinc Ions in Aqueous Solutions


Maize tassel-multiwalled carbon nanotube (MT-MWCNT) composite has been used as a matrix for physical adsorption of horseradish peroxidase (HRP) onto the surface of a glassy carbon electrode through electrostatic interactions. The HRP/MT-MWCNT biosensor was applied for the detection of Zn2+ in aqueous solution. The biosensor designed was able to determine Zn2+ in the range of 0.35 - 12 mg/L with a detection limit of 7.5 μg/L. The inhibition was found to be reversible and uncompetitive when data were modeled using the Dixon and Cornish-Bowden plots. The biosensor was found to have good repeatability, reproducibility and high selectivity. The developed biosensor can be used to detect other HRP inhibiting trace metal ions.

Share and Cite:

Moyo, M. (2014) Horseradish Peroxidase Biosensor to Detect Zinc Ions in Aqueous Solutions. Open Journal of Applied Biosensor, 3, 1-7. doi: 10.4236/ojab.2014.31001.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] R. B. Thompson, B. P. Maliwal, V. L. Feliccia, C. A. Fierke and K. McCall, “Determination of Picomolar Concentrations of Metal Ions Using Fluorescence Anisotropy: Biosensing with a “Reagentless” Enzyme Transducer,” Analytical Chemistry, Vol. 70, No. 22, 1998, pp. 4717-4723.
[2] I. Bontidean, C. Berggren, G. Johansson, et al., “Detection of Heavy Metal Ions at Femtomolar Levels Using Protein-Based Biosensors,” Analytical Chemistry, Vol. 70, No. 19, 1998, pp. 4162-4169.
[3] S. Han, M. Zhu, Z. Yuan and X. Li, “A Methylene Blue-Mediated Enzyme Electrode for the Determination of Trace Mercury(II), Mercury(I), Methylmercury, and Mercuryglutathione Complex,” Biosensors and Bioelectronics, Vol. 16, No. 1-2, 2001, pp. 9-16.
[4] M. R. Guascito, C. Malitesta, E. Mazzotta and A. Turco, Inhibitive Determination of Metal Ions by an Amperometric Glucose Oxidase Biosensor: Study of the Effect of Hydrogen Peroxide Decomposition,” Sensors and Actuators B: Chemical, Vol. 131, No. 2, 2008, pp. 394-402.
[5] E. Ghica, R. C. Carvalho, A. Amine and C. M. A. Brett, “Glucose Oxidase Enzyme Inhibition Sensors for Heavy Metals at Carbon Film Electrodes Modified with Cobalt or Copper Hexacyanoferrate,” Sensors and Actuators B: Chemical, Vol. 178, 2013, pp. 270-278.
[6] S. Rodriguez-Mozaz, M. J. L. De Alda and D. Barcelo, “Biosensors as Useful Tools for Environmental Analysis and Monitoring,” Analytical and Bioanalytical Chemistry, Vol. 386, No. 4, 2006, pp. 1025-1041.
[7] Z. S. Yang, W. L. Wu, X. Chen and Y. C. Liu, “An Amperometric Horseradish Peroxidase Inhibition Biosensor for the Determination of Phenylhydrazine,” Analytical Science, Vol. 24, No. 7, 2008, pp. 895-899.
[8] M. Moyo, J. O. Okonkwo and N. M. Agyei, “A Novel Hydrogen Peroxide Biosensor Based on Adsorption of Horseradish Peroxidase onto a Nanobiomaterial Composite Modified Glassy Carbon Electrode,” Electroanalysis, Vol. 25, No. 8, 2013, pp. 1946-1954.
[9] M. Moyo, L. Chikazaza, B. C. Nyamunda and U. Guyo, “Adsorption Batch Studies on the Removal of Pb(II) Using Maize Tassel Based Activated Carbon,” Journal of Chemistry, Vol. 2013, Article ID: 508934, 8 p.
[10] P. Raghu, T. M. Reddy, B. E. K. Swamy, B. N. Chandrashekar, K. Reddaiah and M. Sreedhar, “Development of AChE Biosensor for the Determination of Methyl Parathion and Monocrotophos in Water and Fruit Juices: A Cyclic Voltammetric Study,” Journal of Electroanalytical Chemistry, Vol. 665, 2012, pp. 76-82.
[11] Z. Li and N. Hu, “Direct Electrochemistry of Heme Proteins in Their Layer-By-Layer Films with Clay Nanoparticles,” Journal of Electroanalytical Chemistry, Vol. 558, 2003, pp. 155-165.
[12] Y. Liu, H. Liu and N. Hu, “Core-Shell Nanocluster Films of Hemoglobin and Clay Nanoparticle: Direct Electro-chemistry and Electrocatalysis,” Biophyscal Chemistry, Vol. 117, No. 1, 2005, pp. 27-37
[13] M. E. Ghica and C. M. A. Brett, “Glucose Oxidase Inhibition in Poly (Neutral red) Mediated Enzyme Biosensors for Heavy Metal De-termination,” Microchimica Acta, Vol. 163, No. 3-4, 2008, pp. 185-193.
[14] X. Wang, S. Xia, J. Zhao, H. Zhao and J. N. Renault, “Inhibitive Determination of Heavy Metal Ions by Conductometric Nitrate Reductase Biosensor,” Chem. Res. Chinese Universities, 2009, Vol. 25, pp. 443-445
[15] F. Arduini, A. Amine, D. Moscone and G. Palleschi, “Reversible Enzyme Inhibition-Based Biosensors: Applica-tions and Analytical Improvement through Diagnostic Inhibition,” Analytical. Letters, Vol. 42, No. 9, 2009, pp. 1258-1293.
[16] G. L. Turdean, “Design and Development of Biosensors for the Detection of Heavy Metal Toxicity,” International Journal of Electrochemistry, Vol. 2011, 2011, pp. 1-15.
[17] A. Cornish-Bowden, “A Simple Graphical Method for Determining the Inhibition Constants of Mixed, Uncompetitive and Non-Competitive Inhibitors,” Biochemical Journal, Vol. 137, 1974, pp. 143-144.
[18] M. Dixon, “The Determination of Enzyme Inhibitor Constants,” Biochemistry Journal. Vol. 55, No. 1, 1953, pp. 170-171.
[19] A. Amine, H. Mohamed, I. Bourais and G. Palleschi, “Enzyme Inhibition-Based Biosensors for Food Safety and Environmental Monitoring,” Biosensors and Bioelectronics, Vol. 21, No. 8, 2006, pp. 1405-1423.
[20] Y. Yang, M. Yang, H. Wang, J. Jiang, G. Shen and R. Yu, “An Amperometric Horseradish Peroxidase Inhibition Biosensor Based on a Cysteamine Self-Assembled Monolayer for the Determination of Sulphides.” Sensors and Actuators B: Chemical, Vol. 102, No. 1, 2004, pp. 162-168.
[21] USEPA, “USEPA Maximum Contaminant Level Goals and National Primary Drinking Water Regulations for Lead and Copper,” Final Rule, Federal Register, Vol. 56, No. 110, 1991, pp. 26460-26469.
[22] World Health Organization (WHO), “Guidelines for Drinking Water Quality,” 3rd Edition, 2004, WHO, Geneva.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.