An amperometric hydrogen peroxide biosensor based on immobilizing horseradish peroxidase to a nano-Au monolayer supported by sol-gel derived carbon ceramic electrode.

A novel strategy for fabricating horseradish peroxidase (HRP)-based H(2)O(2) sensor has been developed by combining the merits of carbon sol-gel supporting matrix and nano-scaled particulate gold (nano-Au) mediator. The thiol functional group-derived carbon ceramic electrode (CCE) was first constructed using (3-mercaptopropyl) trimethoxy silane as sol-gel monomer. Then, the stable nano-Au monolayer was obtained through covalent linkage between nano-Au and thiol group on the surface of CCE. The experimental results showed that nano-Au monolayer formed not only could steadily immobilize HRP but also efficiently retain its bioactivity. Hydrogen peroxide was detected with the aid of hydroquinone mediator to transfer electrons between the electrode and HRP. The process parameters for the fabrication of the enzyme electrode and various experimental variables such as the operating potential, mediator concentration and pH of background electrolyte were explored for optimum analytical performance of the enzyme electrode. The biosensor had a fast response of less than 8 s with linear range of 1.22 x 10(-5) to 1.10 x 10(-3)mol l(-1) and a detection limit of 6.1 x 10(-6)mol l(-1). The sensitivity of the sensor for H(2)O(2) was 0.29 A l mol(-1) cm(-2). The activation energy for enzyme reaction was calculated to be 10.1 kJ mol(-1). The enzyme electrode retained 75% of its initial activity after 5 weeks storage in phosphate buffer at pH 7.

[1]  Juan Li,et al.  Reagentless Amperometric Determination of Hydrogen Peroxide by Silica Sol–Gel Modified Biosensor , 1997 .

[2]  S. Tan,et al.  Amperometric hydrogen peroxide biosensor with silica sol-gel/chitosan film as immobilization matrix , 2001 .

[3]  S. Tan,et al.  Amperometric hydrogen peroxide biosensor based on immobilization of peroxidase in chitosan matrix crosslinked with glutaraldehyde , 2000 .

[4]  A. Desideri,et al.  An electrochemical multienzymatic biosensor for determination of cholesterol. , 2001, Bioelectrochemistry.

[5]  G. Shen,et al.  Renewable amperometric immunosensor for Schistosoma japonium antibody assay. , 2001, Analytical chemistry.

[6]  Ioanis Katakis,et al.  Improved mediated tyrosinase amperometric enzyme electrodes , 1997 .

[7]  María Begoña González-García,et al.  Metal‐Nanoparticles Based Electroanalysis , 2002 .

[8]  J. Wang,et al.  One-step electropolymeric co-immobilization of glucose oxidase and heparin for amperometric biosensing of glucose. , 2000, The Analyst.

[9]  A. L. Crumbliss,et al.  A xanthine oxidase/colloidal gold enzyme electrode for amperometric biosensor applications , 1996 .

[10]  G. E. De Benedetto,et al.  Lactate amperometric biosensor based on an electrosynthesized bilayer film with covalently immobilized enzyme , 1997 .

[11]  P. Southwell-keely,et al.  A reagentless amperometric biosensor for hydrogen peroxide determination based on asparagus tissue and ferrocene mediation , 1995 .

[12]  E. Wang,et al.  Lipid membrane immobilized horseradish peroxidase biosensor for amperometric determination of hydrogen peroxide. , 2003, Biosensors & bioelectronics.

[13]  J R Durrant,et al.  Protein Adsorption on Nanocrystalline TiO(2) Films:  An Immobilization Strategy for Bioanalytical Devices. , 1998, Analytical chemistry.

[14]  Ovadia Lev,et al.  Sol-gel derived renewable-surface biosensors , 1995 .

[15]  A. L. Crumbliss,et al.  Colloidal gold as a biocompatible immobilization matrix suitable for the fabrication of enzyme electrodes by electrodeposition , 1992, Biotechnology and bioengineering.

[16]  N. Uehera,et al.  Determination of hydrogen peroxide based on a metal dispersed sol-gel derived ceramic-graphite composite electrode , 2002, Analytical and bioanalytical chemistry.

[17]  Akira Fujishima,et al.  Tyrosinase-modified boron-doped diamond electrodes for the determination of phenol derivatives , 2002 .

[18]  Xin Wei,et al.  Integration of enzymes and electrodes: spectroscopic and electrochemical studies of chitosan-enzyme films. , 2002, Analytical chemistry.

[19]  O. Lev,et al.  Sol-Gel-Derived Ceramic-Carbon Composite Electrodes: Introduction and Scope of Applications , 1994 .

[20]  M. Natan,et al.  Colloidal Au-enhanced surface plasmon resonance immunosensing. , 1998, Analytical chemistry.

[21]  Joseph Wang,et al.  Metal nanoparticle-based electrochemical stripping potentiometric detection of DNA hybridization. , 2001, Analytical chemistry.

[22]  J. Wang,et al.  Amperometric biosensor for phenols based on a tyrosinase-graphite-epoxy biocomposite. , 1994, The Analyst.

[23]  Shaojun Dong,et al.  Sol-gel-derived titanium oxide/copolymer composite based glucose biosensor. , 2003, Biosensors & bioelectronics.

[24]  J. Storhoff,et al.  Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. , 1997, Science.

[25]  Baohong Liu,et al.  Amperometric biosensor based on a nanoporous ZrO2 matrix , 2003 .

[26]  C. Malitesta,et al.  Glucose fast-response amperometric sensor based on glucose oxidase immobilized in an electropolymerized poly(o-phenylenediamine) film. , 1990, Analytical chemistry.

[27]  Steven R. Emory,et al.  Probing Single Molecules and Single Nanoparticles by Surface-Enhanced Raman Scattering , 1997, Science.

[28]  M. Natan,et al.  MORPHOLOGY-DEPENDENT ELECTROCHEMISTRY OF CYTOCHROME C AT AU COLLOID-MODIFIED SNO2 ELECTRODES , 1996 .

[29]  Haiqing Gong,et al.  Covalent immobilization of an enzyme (glucose oxidase) onto a carbon sol–gel silicate composite surface as a biosensing platform , 2003 .

[30]  Sophie Demoustier-Champagne,et al.  Immobilisation of glucose oxidase within metallic nanotubes arrays for application to enzyme biosensors. , 2003, Biosensors & bioelectronics.

[31]  Frieder W. Scheller,et al.  Enzyme Electrodes Using Bioelectrocatalytic Reduction of Hydrogen Peroxide , 1990 .

[32]  Gang Wang,et al.  Amperometric hydrogen peroxide biosensor with sol-gel/chitosan network-like film as immobilization matrix. , 2003, Biosensors & bioelectronics.

[33]  Yifei Yang,et al.  Bioelectrochemical responses of the polyaniline horseradish peroxidase electrodes , 1997 .

[34]  I. Willner,et al.  Organization of Au Colloids as Monolayer Films onto ITO Glass Surfaces: Application of the Metal Colloid Films as Base Interfaces To Construct Redox-Active Monolayers , 1995 .