A Biosensor Based on Immobilization of Horseradish Peroxidase in Chitosan Matrix Cross-linked with Glyoxal for Amperometric Determination of Hydrogen Peroxide

An amperometric biosensor for hydrogen peroxide (H2O2) was developed via an easy and effective enzyme immobilization method with the “sandwich” configuration: ferrocene-chitosan: HRP: chitosan-glyoxal using a glassy carbon electrode as the basic electrode. In order to prevent the loss of immobilized HRP under optimized conditions, the biosensor surface was cross-linked with glyoxal. Ferrocene was selected and immobilized on the glassy carbon electrode surface as a mediator. The fabrication procedure was systematically optimized to improve the biosensor performance. The biosensor had a fast response of less than 10 s to H2O2, with a linear range of 3.5×10-5 to 1.1×10-3 M, and a detection limit of 8.0×10-6 M based on S/N = 3.

[1]  T. Hianik,et al.  AGAR-SUPPORTED LIPID BILAYERS : BASIC STRUCTURES FOR BIOSENSOR DESIGN. ELECTRICAL AND MECHANICAL PROPERTIES , 1998 .

[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]  Prem C. Pandey,et al.  A novel ferrocene encapsulated palladium-linked ormosil-based electrocatalytic dopamine biosensor , 2001 .

[5]  W. Kaminski,et al.  Separation of Cr(VI) on chitosan membranes , 1999 .

[6]  P C Pandey,et al.  Studies on the electrochemical performance of glucose biosensor based on ferrocene encapsulated ORMOSIL and glucose oxidase modified graphite paste electrode. , 2003, Biosensors & bioelectronics.

[7]  Y. Nakabayashi,et al.  Amperometric Glucose Sensors Fabricated by Electrochemical Polymerization of Phenols on Carbon Paste Electrodes Containing Ferrocene as an Electron Transfer Mediator , 1998 .

[8]  S. Cosnier,et al.  Fabrication of organic phase biosensors based on multilayered polyphenol oxidase protected by an alginate coating , 2001 .

[9]  S. Dong,et al.  Amperometric enzyme electrode for the determination of hydrogen peroxide based on sol–gel/hydrogel composite film , 2000 .

[10]  B. Peppley,et al.  Synthesis, Characterization and Ionic Conductive Properties of Phosphorylated Chitosan Membranes , 2003 .

[11]  Jing-Juan Xu,et al.  Reagentless chemiluminescence biosensor for determination of hydrogen peroxide based on the immobilization of horseradish peroxidase on biocompatible chitosan membrane , 2002 .

[12]  Y. Baba,et al.  Selective Adsorption of Mercury(II) on Chitosan Derivatives from Hydrochloric Acid , 1998 .

[13]  B. González,et al.  A Siloxane Homopolymer with Interacting Ferrocenes as a New Material for the Preparation of Sensors Based on the Detection of Hydrogen Peroxide , 2003 .

[14]  P. Pandey,et al.  Bioelectrochemistry of glucose oxidase immobilized on ferrocene encapsulated ormosil modified electrode , 2001 .

[15]  Shaojun Dong,et al.  Organically Modified Sol‐Gel/Chitosan Composite Based Glucose Biosensor , 2003 .

[16]  M. Costantini,et al.  Influence of glutaraldehyde on drug release and mucoadhesive properties of chitosan microspheres , 1998 .

[17]  J. Magalhães,et al.  Urea potentiometric biosensor based on urease immobilized on chitosan membranes. , 1998, Talanta.

[18]  N. Goh,et al.  Amperometric Glucose Biosensor Based on Immobilization of Glucose Oxidase in Chitosan Matrix Cross-Linked with Glutaraldehyde , 2001 .

[19]  D. Horne,et al.  Influence of whey protein denaturation on κ-carrageenan gelation , 1999 .

[20]  Y. Nakabayashi,et al.  Amperometric Biosensors for Sensing of Hydrogen Peroxide Based on Electron Transfer between Horseradish Peroxidase and Ferrocene as a Mediator , 2000 .

[21]  Y. Baba,et al.  Preparation of Chitosan Derivatives Containing Methylthiocarbamoyl and Phenylthiocarbamoyl Groups and Their Selective Adsorption of Copper(II) over Iron(III) , 2002, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[22]  S. Dong,et al.  Functionalized inorganic–organic composite material derivated by sol–gel for construction of mediated amperometric hydrogen peroxide biosensor , 1999 .

[23]  S. Bao,et al.  Silver-selective Sensor Using an Electrode-separated Piezoelectric Quartz Crystal Modified with a Chitosan Derivative , 2002, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[24]  K. Sugawara,et al.  Electrochemical Sensing of Glucose at a Platinum Electrode with a Chitin/Glucose Oxidase Film , 2000 .

[25]  M. Kawasaki,et al.  Electrode coatings based on chitosan scaffolds. , 2000, Analytical chemistry.

[26]  M. Oshima,et al.  Adsorption Behavior of Metal Ions on Cross-linked Chitosan and the Determination of Oxoanions after Pretreatment with a Chitosan Column , 2000 .

[27]  J. Rhee,et al.  Evaluation of Chitin and Chitosan as a Sorbent for the Preconcentration of Phenol and Chlorophenols in Water , 1998 .

[28]  K. Oshita,et al.  Adsorption Behavior of Mercury and Precious Metals on Cross-Linked Chitosan and the Removal of Ultratrace Amounts of Mercury in Concentrated Hydrochloric Acid by a Column Treatment with Cross-Linked Chitosan , 2002, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[29]  Xiaogang Wu,et al.  Studies on Electrochemical Behavior of Bromideat a Chitosan‐Modified Glassy Carbon Electrode , 2001 .

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