Microchimica Acta

, Volume 183, Issue 5, pp 1625–1632 | Cite as

A glassy carbon electrode modified with a composite consisting of reduced graphene oxide, zinc oxide and silver nanoparticles in a chitosan matrix for studying the direct electron transfer of glucose oxidase and for enzymatic sensing of glucose

  • Zhenjiang Li
  • Liying Sheng
  • Alan MengEmail author
  • Cuicui Xie
  • Kun Zhao
Original Paper


The authors describe the fabrication of a nanocomposite consisting of reduced graphene oxide, zinc oxide and silver nanoparticles by microwave-assisted synthesis. The composite was further reduced in-situ with hydrazine hydrate and then placed, along with the enzyme glucose oxidase, on a glassy carbon electrode. The synergistic effect of the materials employed in the nanocomposite result in excellent electrocatalytic activity. The Michaelis-Menten constant of the adsorbed GOx is 0.25 mM, implying a remarkable affinity of the GOx for glucose. The amperometric response of the modified GCE is linearly proportional to the concentration of glucose in 0.1 to 12.0 mM concentration range, and the detection limit is 10.6 µM. The biosensor is highly selective, well reproducible and stable.

Graphical abstract

A nanocomposite consisting of graphene oxide, zinc oxide and silver nanoparticles was prepared by microwave-assisted synthesis and further reduced with 85 % hydrazine hydrate (HAA). The material was incorporated, along with glucose oxidase, into a chitosan (CS) matrix on a glassy carbon electrode to give a glucose biosensor with a 10.6 µM detection limit.


Biosensor Bioassay Enzymatic assay Nanocomposite Microwave synthesis In-situ reduction Hydrazine reduction Direct electrochemistry Amperometry 



The work reported here was supported by the National Natural Science Foundation of China under Grant No.51572137, 51502149, 51272117, 51172115, the Natural Science Foundation of Shandong Province under Grant No.ZR2015PE003, ZR2013EMQ006, the Research Award Fund for Outstanding Young Scientists of Shandong Province Grant No. BS2013CL040, the Specialized Research Fund for the Doctoral Program of Higher Education of China under Grant No. 20123719110003, the Tackling Key Program of Science and Technology in Shandong Province under Grant No. 2012GGX1021, the Application Foundation Research Program of Qingdao under Grant No. 13-1-4-117-jch, 15-9-1-28-jch, 14-2-4-29-jch, Shandong Province Taishan Scholar Project. We express our grateful thanks to them for their financial support.

Compliance with ethical standards must be captured

The author(s) declare that they have no competing interests

Supplementary material

604_2016_1791_MOESM1_ESM.doc (556 kb)
ESM 1 (DOC 556 kb)


  1. 1.
    Karyakin AA (2012) Principles of direct (mediator free) bioelectrocatalysis. Bioelectrochemistry 88:70–75CrossRefGoogle Scholar
  2. 2.
    Habermüller K, Mosbach M, Schuhmann W (2000) Electron-transfer mechanisms in amperometric biosensors. Fresenius J Anal Chem 366:560–568CrossRefGoogle Scholar
  3. 3.
    Gu M, Wang J, Tu Y, Di J (2010) Fabrication of reagentless glucose biosensors: a comparison of mono-enzyme GOD and bienzyme GOD-HRP systems. Sensors Actuators B Chem 148:486–491CrossRefGoogle Scholar
  4. 4.
    Deng CY, Chen JH, Nie Z, Si SH (2010) A sensitive and stable biosensor based on the direct electrochemistry of glucose oxidase assembled layer-by-layer at the multiwall carbon nanotube-modified electrode. Biosens Bioelectron 26:213–219CrossRefGoogle Scholar
  5. 5.
    Wilson R, Turner APF (1992) Glucose oxidase: an ideal enzyme. Biosens Bioelectron 7:165–185CrossRefGoogle Scholar
  6. 6.
    Holland JT, Lau C, Brozik S, Atanassov P, Banta S (2011) Engineering of glucose oxidase for direct electron transfer via site-specific gold nanoparticle conjugation. J Am Chem Soc 133:19262–19265CrossRefGoogle Scholar
  7. 7.
    Su S, Sun HF, Xu F, Yuwen LH, Fan CH, Wang LH (2014) Direct electrochemistry of glucose oxidase and a biosensor for glucose based on a glass carbon electrode modified with MoS2 nanosheets decorated with gold nanoparticles. Microchim Acta 181:1497–1503CrossRefGoogle Scholar
  8. 8.
    Karuppiah C, Palanisamy S, Chen SM, Veeramani V, Periakaruppan P (2014) Direct electrochemistry of glucose oxidase and sensing glucose using a screen-printed carbon electrode modified with graphite nanosheets and zinc oxide nanoparticles. Microchim Acta 181:1843–1850CrossRefGoogle Scholar
  9. 9.
    Liu S, Tian J, Wang L, Luo Y, Lu W, Sun X (2011) Self-assembled graphene platelet-glucose oxidase nanostructures for glucose biosensing. Biosens Bioelectron 26:4491–4496CrossRefGoogle Scholar
  10. 10.
    Li D, Muller B, Gilje S, Kaner RB, Wallace GG (2008) Processable aqueous dispersions of graphene nanosheets. Nat Nanotechnol 3:101–105CrossRefGoogle Scholar
  11. 11.
    Wang F, Gong WC, Wang LL, Chen ZL (2015) Enhanced amperometric response of a glucose oxidase and horseradish peroxidase based bienzyme glucose biosensor modified with a film of polymerized toluidine blue containing reduced graphene oxide. Microchim Acta 182:1949–1956CrossRefGoogle Scholar
  12. 12.
    Kang XH, Wang J, Wu H, Ilhan AA, Liu J, Lin YH (2009) Glucose oxidase-graphene-chitosan modified electrode for direct electrochemistry and glucose sensing. Biosens Bioelectron 25:901–907CrossRefGoogle Scholar
  13. 13.
    Wang CY, Tan XR, Chen SH, Yuan R, Hu FX, Yuan DH, Xiang Y (2012) Highly-sensitive cholesterol biosensor based on platinum-gold hybrid functionalized ZnO nanorods. Talanta 94:263–270CrossRefGoogle Scholar
  14. 14.
    Li YF, Liu ZM, Liu YL, Yang YH, Shen GL, Yu RQ (2006) A mediator-free phenol biosensor based on immobilizing tyrosinase to ZnO nanoparticles. Anal Biochem 349:33–40CrossRefGoogle Scholar
  15. 15.
    Zhang F, Wang X, Ai S, Sun Z, Wan Q, Zhu Z, Xian Y, Jin L, Yamamoto K (2004) Immobilization of uricase on ZnO nanorods for a reagentless uric acid biosensor. Anal Chim Acta 519:155–160CrossRefGoogle Scholar
  16. 16.
    Wang Q, Zheng JB (2010) Electrodeposition of silver nanoparticles on a zinc oxide film: improvement of amperometric sensing sensitivity and stability for hydrogen peroxide determination. Microchim Acta 169:361–365CrossRefGoogle Scholar
  17. 17.
    Ahmad M, Pan C, Gan L, Nawaz ZS, Zhu J (2010) Highly sensitive amperometric cholesterol biosensor based on Pt-Incorporated fullerene-like ZnO nanospheres. J Phys Chem C 114:243–250CrossRefGoogle Scholar
  18. 18.
    Norouzi P, Ganjali H, Larijani B, Ganjali MR, Faridbod F, Zamani HA (2011) A glucose biosensor based on nanographene and ZnO nanoparticles using FFT Continuous cyclic voltammetry. Int J Electrochem Sci 6:5189–5199Google Scholar
  19. 19.
    Huang JY, Zhao MG, Ye ZZ (2014) Electrospun porous ZnO nanofibers for glucose biosensors. Adv Mater Res 950:3–6CrossRefGoogle Scholar
  20. 20.
    Ali USM, Nur O, Willander M, Danielsson B (2009) Glucose detection with a commercial MOSFET using a ZnO nanowires extended gate. IEEE Transactions on Nanotechnology 8:678–683CrossRefGoogle Scholar
  21. 21.
    Li ZJ, Xie CC, Wang JH, Meng AL (2015) Direct electrochemistry of cholesterol oxidase immobilized on chitosan–grapheme and cholesterol sensing. Sensors Actuators B 208:505–511CrossRefGoogle Scholar
  22. 22.
    Meng AL, Shao J, Fan XY, Wang JH (2014) Rapid synthesis of a flower-like ZnO/rGO/Ag micro/nano-composite with enhanced photocatalytic performance by a one-step microwave method. RSC Adv 4:60300–60305CrossRefGoogle Scholar
  23. 23.
    Yang D, Velamakanni A, Bozoklu G, Park S, Stoller M, Piner RD, Stankovich S, Jung I, Field DA, Ventrice CA, Ruoff RS (2009) Chemical analysis of graphene oxide films after heat and chemical treatments by X-ray photoelectron and micro-Raman spectroscopy. Carbon 47:145–152CrossRefGoogle Scholar
  24. 24.
    Liu YG, Feng XM, Shen JM, Zhu JJ (2008) Fabrication of a novel glucose biosensor based on a highly electroactive polystyrene/polyaniline/Au nanocomposite. J Phys Chem B 112:9237–9242CrossRefGoogle Scholar
  25. 25.
    Battaglini F, Bartlett PN, Wang JH (2000) Covalent attachment of osmium complexes to glucose oxidase and the application of the resulting modified enzyme in an enzyme switch responsive to glucose. Anal Chem 72:502–509CrossRefGoogle Scholar
  26. 26.
    Shan D, Zhang J, Xue HG, Ding SN, Cosnier S (2010) Colloidal laponite nanoparticles: extended application in direct electrochemistry of glucose oxidase and reagentless glucose biosensing. Biosens Bioelectron 25:1427–1433CrossRefGoogle Scholar
  27. 27.
    Laviron E (1979) General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems. J Electroanal Chem 101:19–28CrossRefGoogle Scholar
  28. 28.
    Wu P, Shao Q, Hu YJ, Jin J, Yin YJ, Zhang H (2010) Direct electrochemistry of glucose oxidase assembled on graphene and application to glucose detection. Electrochim Acta 55:8606–8614CrossRefGoogle Scholar
  29. 29.
    Xiao XL, Zhou B, Zhu L, Xu LL, Tan L, Tang H (2012) An reagentless glucose biosensor based on direct electrochemistry of glucose oxidase immobilized on poly (methylene blue) doped silica nanocomposites. Sensors Actuators B Chem 165:126–132CrossRefGoogle Scholar
  30. 30.
    Zhu L, Xu LL, Tan L, Tan H (2013) Direct electrochemistry of cholesterol oxidase immobilized on gold nanoparticles-decorated multiwalled carbon nanotubes and cholesterol sensing. Talanta 106:192–199CrossRefGoogle Scholar
  31. 31.
    Scott DL, Bowden EF (1994) Enzyme-substrate kinetics of adsorbed cytochrome c peroxidase on pyrolytic graphite electrodes. Anal Chem 66:1217–1223CrossRefGoogle Scholar
  32. 32.
    Jiang HD, Kim SK (2012) A glucose biosensor based on TiO2-graphene composite. Biosens Bioelectron 38:184–188CrossRefGoogle Scholar
  33. 33.
    Chen XL, Chen JH, Deng CY, Xiao CH, Yang YM, Nie Z (2008) Amperometric glucose biosensor based on boron-doped carbon nanotubes modified electrode. Talanta 76:763–767CrossRefGoogle Scholar
  34. 34.
    Oztekin Y, Ramanaviciene A, Yazicigil Z, Solak AO, Ramanavicius A (2011) Direct electron transfer from glucose oxidase immobilized on polyphenanthroline-modified glassy carbon electrode. Biosens Bioelectron 26:2541–2546CrossRefGoogle Scholar
  35. 35.
    Du Y, Luo XL, Xu JJ, Chen HY (2007) A simple method to fabricate a chitosan-gold nanoparticles film and its application in glucose biosensor. Bioelectrochem 70:342–347CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  • Zhenjiang Li
    • 1
    • 2
  • Liying Sheng
    • 1
  • Alan Meng
    • 2
    Email author
  • Cuicui Xie
    • 1
  • Kun Zhao
    • 2
  1. 1.Key Laboratory of Polymer Material Advanced Manufacturings Technology of Shandong Provincial, College of Electromechanical Engineering, College of Sino-German Science and TechnologyQingdao University of Science and TechnologyQingdaoPeople’s Republic of China
  2. 2.State Key Laboratory Base of Eco-chemical Engineering, College of Chemistry and Molecular EngineeringQingdao University of Science and TechnologyQingdaoPeople’s Republic of China

Personalised recommendations