CONSTRUCTION OF AN ELECTROCHEMICAL XANTHINE BIOSENSOR BASED ON GRAPHENE/COBALT OXIDE NANOPARTICLES/CHITOSAN COMPOSITE FOR FISH FRESHNESS DETECTION

Abstract

Xanthine biosensor based on glassy carbon electrode (GC) modified with graphene (GR), Co₃O₄ nanoparticles and chitosan (CH) composite was fabricated. Xanthine oxidase (XAO) solution was dropped on the surface of Co₃O₄/CH/GR/GC and electrode was placed in saturated glutaraldehyde vapor for the crosslinking of the enzymes. The modified electrode was characterized by scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy. Under the optimized experimental conditions, xanthine was detected in the concentration range from 5.0×10^-7 to 8.0×10^-5 M with a detection limit of 2.0×10^-7 M. The low Michaelis–Menten constant (0.17 mM) suggested enhanced enzyme affinity for the immobilized enzyme as compared to previously reported xanthine biosensors. Moreover, the biosensor exhibited some advantages, such as short response time (10 s), high sensitivity (6.58 μA/mM or 74.8 µA/mMcm²) and good reproducibility (RSD = 1.2%). The suitability of the device was verified by xanthine assay in a fish meat. The biosensor provide to be a reliable, easy, fast and economical method for the evaluation of fish freshness.

References

1. Niu J, Lee, JY. A new approach for the determination of fish freshness by electrochemical impedance spectroscopy. J. Food Sci. 2000;65(5):780-785.
2. Anderson AK. Biogenic and volatile amine-related qualities of three popular fish species sold at Kuwait fish markets. Food Chem. 2000;107(2):761‒767.
3. Ota F, Nakamura T. Variation of ammonia contents in fish meat by heating under pressure: relation between the increase of ammonia and the freshness of fish. Bull. Japan. Soc. Sci. Fish. 1952;18:15–20.
4. Suzuki T. Determination of volatile acids for judging the freshness of fish. Bull. Japan. Soc. Sci. Fish. 1953;19:102–106.
5. Kawabata T, Fujimaki M, Amano K, Tomiya F. Studies of the pH for fish muscle. Bull. Japan. Soc. Sci. Fish. 1952;18:124–132.
6. Nakatani HS, Vieira dos Santos L, Pelegrine CP, Gomes STM, Matsushita M, Evelázio de Souza NJ, Visentainer V. Biosensor based on xanthine oxidase for monitoring hypoxanthine in fish meat. Am. J. Biochem. Biotechnol. 2005;1:85‒89.
7. Lawal AT, Adeloju SB. Progress and recent advances in fabrication and utilization of hypoxanthine biosensors for meat and fish quality assessment: A review. Talanta 2012;100:217–228.
8. Pundir CH, Devi R. Biosensing methods for xanthine determination: A review. Enzyme Microb. Tech. 2014;57:55–62.

9. Pagliarussi RS, Freitas LAP, Bastos JK. A quantitative method for the analysis of xanthine alkaloids in Paullinia cupana (guarana) by capillary column gas chromatography. J. Sep. Sci. 2002;25:371−374.
10. Cooper N, Khosravan R, Erdmann C, Fiene J, Lee JW. Quantification of uric acid, xanthine and hypoxanthine in human serum by HPLC for pharmacodynamic studies. J. Chrom. B. 2006;837:1–10.
11. Hlavay J, Haemmerli S, Gailbult G. Fibre-optic biosensor for hypoxanthine and xanthine based on a chemiluminescence reaction. Biosens. Bioelectron. 1994;9:189-195.
12. Amigo JM, Coello J, Maspoch S. Three-way partial least-squares regression for the simultaneous kinetic-enzymatic determination of xanthine and hypoxanthine in human urine. Anal. Bioanal. Chem. 2005;382:1380–1388.
13. Dervisevic M, Custiuc E, Çevik E, Durmuş Z, Şenel M, Durmuş A. Electrochemical biosensor based on REGO/Fe3O4 bionanocomposite interface for xanthine detection in fish sample. Food Control 2015; 57:402−410.
14. Dalkiran B, Kacar C, Erden PE, Kilic E. Amperometric xanthine biosensors based on chitosan-Co3O4-multiwall carbon nanotube modified glassy carbon electrode. Sensor. Actuat. B:Chem. 2014;200:83‒91.
15. [
16. Villalonga R, Diez P, Gamella M, Reviejo J, Pingarrón JM. Immobilization of xanthine oxidase on carbon nanotubes through double supramolecular junctions for biosensor construction. Electroanal. 2011;23(8):1790-1796.
17. Oztürk FÖ, Erden, PE, Kacar C, Kilic E. Amperometric biosensor for xanthine determination based on Fe3O4 nanoparticles. Acta Chim. Slov. 2013;61(1):19‒26.
18. Chen J, Zheng X, Miao F, Zhang J, Cui X, Zheng W. Engineering graphene/carbon nanotube hybrid for direct electron transfer of glucose oxidase and glucose biosensor. J. Appl. Electrochem. 2012;42:875-881.
19. Kuila T, Bose S, Khanra P, Mishra AK, Kim NH, Lee JH (2011). Recent advances in graphene-based biosensors. Biosens. Bioelectron. 2011;26(12):4637‒4648.
20. Zhou K, Zhu Y, Yang X, Li C. Preparation and application of mediator‐free H2O2 biosensors of graphene‐Fe3O4 composites. Electroanal. 2011;23(4):862-869.
21. Fei J, Wen J, Yi L, Ge F, Zhang Y, Huang M, Chen X.
22. Electrochemical determination diethylstilbestrol by a single-walled carbon nanotube/platinum nanoparticle composite film electrode. J. Appl. Electrochem. 2008;38:1527–1533.
23. Kacar C, Dalkiran B, Erden PE, Kilic E. An amperometric hydrogen peroxide biosensor based on Co3O4 nanoparticles and multiwalled carbon nanotube modified glassy carbon electrode. Appl. Surf. Sci. 2014;311:139‒146.
24. Zeng Q, Cheng JS, Liu XF, Bai HT, Jiang JH. Palladium nanoparticle/chitosan-grafted graphene nanocomposites for construction of a glucose biosensor. Biosens. Bioelectron. 2011;26(8):3456-3463.
25. Teymourian H, Salimi A, Khezrian S. Fe3O4 magnetic nanoparticles/reduced graphene oxide nanosheets as a novel electrochemical and bioelectrochemical sensing platform. Biosens. Bioelectron. 2013;49:1–8.
26. Fu L, Zheng YH, Fu ZX. Ascorbic acid amperometric sensor using a graphene-wrapped hierarical TiO2 nanocomposite. Chem. Pap. 2015;69:655-661.
27. Wu H, Wang J, Kang X, Wang C, Wang D, Liu J, Aksay IA, Lin Y. Glucose biosensor based on immobilization of glucose oxidase in platinum nanoparticles/graphene/chitosan nanocomposite film. Talanta 2009;80:403–406.
28. Zhou K, Zhu Y, Yang X, Luo J, Li C, Luan S. A novel hydrogen peroxide biosensor based on Au–graphene–HRP–chitosan biocomposites. Electrochim. Acta 2010;55:3055–3060.
29. Chuah AM, Kuroiwa T, Ichikawa S, Kobayash I, Nakajima M. Formation of biocompatible nanoparticles via the self-assembly of chitosan and modified lecithin. J. Food Sci. 2008;74(1):1-8.
30. Zargar B, Parhamand H, Hatamie A. Electrochemical investigation and stripping voltammetric determination of captopril at CuO nanoparticles/multi-wall carbon nanotube nanocomposite electrode in tablet and urine samples. Anal. Methods 2015;7:1026‒1035.
31. Pérez-López B, Merkoci A. Carbon nanotubes and graphene in analytical sciences. Microchim. Acta 2012;179(1-2):1‒16.
32. Dong XC, Hang X, Wang XW, Huang YX, Chan-Park MB, Zhang H, Wang LH, Huang W, Chen P. 3D graphene–cobalt oxide electrode for high-performance supercapacitor and enzymeless glucose detection. ACS Nano 2012;6(4): 3206‒3213.
33. Gokoglan TC, Soylemez S, Kesik M, Toksabay S, Toppare L. Selenium containing conducting polymer based pyranose oxidase biosensor for glucose detection. Food Chem. 2015;172: 219‒224.
34. Qi X, Gao H, Zhang Y, Wang X, Chen Y, Sun W. Electrochemical DNA biosensor with chitosan-Co3O4 nanorod-graphene composite for the sensitive detection of staphylococcus aureus nuc gene sequence. Bioelectrochem. 2012;88:42‒47.
35. Shan D, Wang Y, Xue H, Cosnier S. Sensitive and selective xanthine amperometric sensors based on calcium carbonate nanoparticles. Sensor. Actuat. B:Chem. 2009;136:510–515.
36. Arslan F, Yaşar A, Kilic E. An amperometric biosensor for xanthine determination prepared from xanthine oxidase immobilized in polypyrrole film. Artif. Cell Blood Sub. 2006;34(1):111−126.
37. Dodevska T, Horozova E, Dimcheva N. Design of an amperometric xanthine biosensor based on a graphite transducer patterned with noble metal microparticles. Cent. Eur. J. Chem. 2010;8(1):19–27.
38. Devi R, Batra B, Lata S, Yadav S, Pundir CS. A method for determination of xanthine in meat by amperometric biosensor based on silver nanoparticles/cysteine modified Au electrode. Process Biochem. 2013;48,:242–249.
39. Dervisevic M, Custiuc E, Çevik E, Şenel M. Construction of novel xanthine biosensor by using polymeric mediator/MWCNT nanocomposite layer for fish freshness detection. Food Chem. 2015;181:277-283.
40. Pei J, Li XY. Xanthine and hypoxanthine sensors based on xanthine oxidase immobilized on a CuPtCl6 emically modified electrode and liquid chromatography electrochemical detection. Anal. Chim. Acta 2000;414:205–213.
41. [
42. Bas SZ, Gulce H, Yıldız S. Amperometric xanthine biosensors based on electrodeposition of platinum on polyvinylferrocenium coated Pt electrode. J. Mol. Catal. B: Enzym. 2011;7:282–288.
43. Devi R, Thakur M, Pundir C.S. Construction and application of an amperometric xanthine biosensor based on zinc oxide nanoparticles–polypyrrole composite film. Biosens. Bioelectron. 2011;26(8):3420-3426.
44. Jain U, Narang J, Rani K, Chauhan N. Synthesis of cadmium oxide and carbon nanotube based nanocomposites and their use as a sensing interface for xanthine detection. RSC Advances 2015;5(38):29675-29683.