Using Response Surface Methodology for Amperometric Glucose Biosensor Construction

Abstract

In this study, construction of amperometric glucose biosensor was carried out by immobilizing of glucose oxidase (GOD) on platinum electrode with 0.09 cm2 surface area which coated with polypyrrole (PPy) by cyclic voltammetry technique. Because measured current values in the presence of glucose would be affected from the electrode preparing and working conditions, experimental parameters should be optimized by response surface methodology (RSM). To this, State Ease Design Expert 8.0.7.1. (Serial Number:0021-6578) programe was used. PPy synthesis conditions of pyrrole (Py) monomer concentration and scan rate were optimized according to current response in presence of glucose. Optimal Py monomer concentration and scan rate for PPy synthesis were determined as 10 mM and 50 mV/s, respectively. Immobilization parameters such as concentrations of chitosan, GOD and glutaraldehyde (GAL) also were optimized by RSM as 1.0 %, 4 mg/ml and 0.0625 %, respectively. The digital photos of electrodes at each stage were obtained. All electrodes well characterized in absence and in the presence of glucose by cyclic voltammetry and impedance techniques and it was observed that electrodes were sensitive to glucose molecule. Finally the effect of working pH and applied potential on the current response was investigated by RSM. The highest current response was obsreved when pH of glucose solution and applied potential were 6.0 and 0.8, respectively.

Keywords

• Response surface methodology,Pt electrode,Polypyrrole,amperometric biosensor,glucose oxidase

References

1. Ahuja, T., Mir, I.A., Kumar, D. & Rajesh, (2007). Biomolecular immobilization on conducting polymers for biosensing applications. Biomaterials, 28, 791-805.
2. Bal, Ö. (2012) .Preparation of a new biosensor for determination of glucose with immobilization on the polypyrrole film of glucose oxidase in biologically fluids. MSc. Thesis, Gazi University.
3. Bezerra, M.A., Santelli, R.E., Oliveira, E.P., Villar L.S. & Escaleira, L.A. (2013). Highly selective electrochemical biosensor for the determination of folic acid based on DNA modified-pencil graphite electrode using response surface methodology, Materials Science and Engineering C, 33 1753–1758.
4. Borole, D. D., Kapadi, U.R., Mahulikar, P.P. & Hundiwale, D.G. (2004). Glucose Oxidase Electrodes of Polyaniline, Poly(o-toluidine) and Their Copolymer as Biosensor: a Comparative Study. Polymer Advantage Technology, 15, 306-312.
5. Chaichi, M.J. & Ehsani, M. (2016). A novel glucose sensor based on immobilization of glucose oxidase on the chitosan-coated Fe3O4 nanoparticles and the luminol-H2O2-gold nanoparticle chemiluminescence detection system, Sensors and Actuators B-Chemical, 223, 713-722.
6. Chen, C., Jian, Y. & Kan, J. (2005). A noninterference polypyrole glucose biosensor. Biosensors Bioelectronics. 22, 639-643.
7. Dong Y.P., Huang L., Chu X.F. & Pei L.Z. (2013). An Amperometric Glucose Biosensor Based on the Immobilization of Glucose Oxidase on the CuGeO Nanowire Modified Electrode Russian Journal of Electrochemistry, 49(6), 571–576.
8. Ebrahimi, B., Shojaosadati, S.A., Daneshgar, P., Norouzi, P. & Mousavi S.M. (2011). Performance evaluation of fast Fourier-transform continuous cyclic-voltammetry pesticide biosensor Analytica Chimica Acta 687 168–176.

9. Gouda, M. D., Thakur, M. S. & Karanth, N. G. (2001). Optimization of the multienzym system for sucrose biosensor by response surface methodology. World Journal of Microbiology & Biotechnology, 17, 595-600.
10. Haighi, B., Nazari, L. & Sajjadi, S.M., (2012). Fabrication and application of a sensitive and highly stable copper hexacyanoferrate modified carbon ionic liquid paste electrode for hydrogen peroxide and glucose detection, Electroanalysis, 24, 2165-2175.
11. Ivory, D.M., Miller, G.G., Sowa, J.M., Shacklette, L.W., Chance,R.R. & Baughman, R.H. (1979). Highly conducting chargetransfer complexes of poly(p-phenylene). The Journal of Chemical Physics, 71, 1506–1507.
12. Kawai, T., Kuwabara, T., Wang, S. & Yoshino, K. (1990). Secondar battery characteristics poly(3-alkylthiophene). Japanese Journal of Applied Physics, 29, 602–605.
13. Kergaravat, S.V., Pividori, M.I. & Hernandez, S.H. (2012). Evaluation of seven cosubstrates in the quantification of horseradish peroxidase enzyme by square wave voltammetry, Talanta. 88, 468– 476.
14. Jie, Z., Xiaoqiang, L., Xinhai, W. Xiaohe H. & Rui, Y. (2015). Preparation of polyaniline-TiO2 nanotube composite for the development of electrochemicalbiosensors. Sensors and Actuators B-Chemıcal, 221, 450-45.
15. Ma, M., Qu, L. & Shi, G. (2003). Glucose Oxidase Electrodes Based on Microstructured Polypyrole Films. Journal of Applied Polymer Science, 98, 2550-2554.
16. Miyamoto, S., De Taxis du Poet, P., Murakami, T., Kimura, J. & Karube, I. (1990). Direct electron transfer with glucose oxidase immobilized in an electropolymerized poly-N-Methylpyrrole film on gold microelectrode. Analytica Chimica Acta, 235, 255-264.
17. Mirmoghtadaie, L., Ensafi, A.A., Kadivar, M. & Norouzi, P. (2013) Highly selective electrochemical biosensor for the determination of folic acid based on DNA modified-pencil graphite electrode using response surface methodology, Materials Science and Engineering C 33, 1753–1758.
18. Ozyilmaz, G, Tukel, S.S. & Alptekin O. (2005) Activity and storage stability of immobilized glucose oxidase onto magnesium silicate. J Mol Catal B: Enzymatic, 35, 154–60.
19. Ozyilmaz, G., Ozyilmaz, A.T. & Can, F. (2011). Glucose Oxidase- Polypyrrole Electrodes Synthesized In P-Toluenesulfonic Acid And Sodium p-Toluenesulfonate, Applied Biochemistry and Microbiology, 47, 217-225.
20. Ozyilmaz, G., Özyılmaz, A.T., Akyürekoğlu, R. (2017). Poly(N-Methylpyrrole)-Chitosan layers for Glucose Oxidase Immobilization for Amperometric Glucose Biosensor Design. Natural and Engineering Sciences, 2(3): 123-134.
21. Özyılmaz A.T., Avsar B., Ozyilmaz G., Karahan İ.H., Çamurcu T & Çolak F. (2014) .Different copolymer films on ZnFeCo particles: Synthesis and anticorrosion properties, Applied Surface Science, 318, 262-268.
22. Shan D., Wang S., He Y.& Xue H. (2008) Amperometric glucose biosensor based on in situ electropolymerized polyaniline/poly(acrylonitrile-co-acrylic acid) composite film. Materials Science and Engineering, C 28, 213–217.
23. Sung, W.J., Bae, Y. H. (2003) .A glucose oxidase electrode based on polypyrrole with polyanion/PEG/enzyme conjugate dopant. Biosensors and Bioelectronics, 18, 1231-1239.
24. Urkut, Z., Kara, P., Goksungur, Y. & Ozsoz, M. (2011). Response Surface Methodology for Optimization of Food Borne Pathogen Detection in Real Samples Based on Label Free Electrochemical Nucleic Acid Biosensors. Electroanalysıs, 23, 2668-2676.
25. Xu G., Adeloju S.B.O., Wu Y. & Zhang X. (2012). Modification of polypyrrole nanowires array with platinum nanoparticles and glucose oxidase for fabrication of a novel glucose biosensor. Analytica Chimica Acta, 755, 100-107.
26. Zejun, C. Huiling, L., , Xufei, Z., Dong L., Shaoyu Z., Xiaoyuan, C. & Ye, S., 2014. Electropolymerization of Aniline onto Anodic WO3 Film: An Approach to Extend Polyaniline Electroactivity Beyond pH 7. Journal of Physical Chemistry, C.118: 27449-27458.
27. Zhang, Q., Qu, Y., Zhou, J., Zhang, X., Zhou, H., Ma, Q., Li X. (2011). Optimization of 2,3-dihydroxybiphenyl 1,2-dioxygenase expression and its application for biosensor. Bioresource Technology, 102, 10553–10560.