A reduced graphene oxide based biosensor for high-sensitive detection of phenols in water samples

Abstract Laccase (Lac) from Rhus vernificer a was covalently immobilized onto 1-aminopyrene (1-AP) functionalized reduced graphene oxides (rGOs)-modified glassy carbon electrode by encapsulation with chitosan. The biosensor was characterized with respect to the optimum pH and polarized potentials for the determination of phenols, and its response time, sensitivity, linear range, detection limit, and stability. Hydroquinone and catechol were selected as the analytes and detected based on the direct electron transfer behavior of Lac and its enzymatic oxidation of analyte. The sensitivities of the electrode were 14.16, 15.79 μA mM −1 with linear ranges of 3–2000, 15–700 μM for hydroquinone and catechol, respectively. The detection limits (S/N = 3) were 2 and 7 μM for hydroquinone and catechol, respectively with a fast response time ( t 95% ) of less than 5 s. Moreover, the immobilized Lac showed high affinity to hydroquinone and catechol with K m app values of 5 and 0.3 mM, respectively. As a consequence, the biosensor demonstrated suitable stability ( ca. 7 days; over 300 determinations) and good repeatability with a relative standard deviation of 3.96%. The recovery study of hydroquinone in real water samples gave values from 82.7% ± 10% to 105.9% ± 8%, confirming its application potential in the measurement of phenols in real samples.

[1]  Sang Yup Lee,et al.  Solution chemistry of self-assembled graphene nanohybrids for high-performance flexible biosensors. , 2010, ACS nano.

[2]  O. Fatibello‐Filho,et al.  Biosensor based on laccase immobilized on microspheres of chitosan crosslinked with tripolyphosphate , 2008 .

[3]  S. Sario,et al.  Comparison of Immobilisation Procedures for Development of an Electrochemical PPO-Based Biosensor for on Line Monitoring of a Depuration Process , 1994 .

[4]  Y. Chai,et al.  Immobilization of horseradish peroxidase on chitosan/silica sol-gel hybrid membranes for the preparation of hydrogen peroxide biosensor. , 2008, Journal of biochemical and biophysical methods.

[5]  Gabriel Lucian Radu,et al.  Laccase–MWCNT–chitosan biosensor—A new tool for total polyphenolic content evaluation from in vitro cultivated plants , 2010 .

[6]  S. Zuo,et al.  Direct electron transfer of Horseradish peroxidase on porous structure of screen-printed electrode. , 2009, Biosensors & bioelectronics.

[7]  Edward I. Solomon,et al.  ELECTRONIC STRUCTURES OF ACTIVE SITES IN COPPER PROTEINS : CONTRIBUTIONS TO REACTIVITY , 1992 .

[8]  C S Pundir,et al.  Polyphenol biosensor based on laccase immobilized onto silver nanoparticles/multiwalled carbon nanotube/polyaniline gold electrode. , 2011, Analytical biochemistry.

[9]  R. Zengerle,et al.  Carbon electrodes for direct electron transfer type laccase cathodes investigated by current density-cathode potential behavior. , 2010, Biosensors & bioelectronics.

[10]  S. Shleev,et al.  Direct electron transfer reactions of laccases from different origins on carbon electrodes. , 2005, Bioelectrochemistry.

[11]  Lo Gorton,et al.  Direct electron transfer kinetics in horseradish peroxidase electrocatalysis. , 2007, The journal of physical chemistry. B.

[12]  J. Dupont,et al.  Biosensor based on platinum nanoparticles dispersed in ionic liquid and laccase for determination of adrenaline , 2009 .

[13]  Yufeng Zheng,et al.  A comparative study on electrochemistry of laccase at two kinds of carbon nanotubes and its application for biofuel cell , 2008 .

[14]  G. Hicks,et al.  The Enzyme Electrode , 1967, Nature.

[15]  B. Långström,et al.  Cyclic Voltammetry and Electrocatalysis of the Blue Copper Oxidase Polyporus versicolor Laccase. , 1998 .

[16]  Kang Wang,et al.  Application of thermally reduced graphene oxide modified electrode in simultaneous determination of dihydroxybenzene isomers , 2012 .

[17]  E. Ferapontova,et al.  Activation of laccase bioelectrocatalysis of O2 reduction to H2O by carbon nanoparticles , 2012 .

[18]  H. Gray,et al.  Catalysis of the reduction of dioxygen at graphite electrodes coated with fungal laccase A , 1984 .

[19]  A. Yaropolov,et al.  Laccase: properties, catalytic mechanism, and applicability , 1994 .

[20]  J. Rogalski,et al.  Powerful connection of laccase and carbon nanotubes: Material for mediator-free electron transport on the enzymatic cathode of the biobattery , 2012 .

[21]  T. E. Abraham,et al.  Biosensor for the determination of phenols based on cross-linked enzyme crystals (CLEC) of laccase. , 2005, Biosensors & bioelectronics.

[22]  Ming Zhou,et al.  Controlled synthesis of large-area and patterned electrochemically reduced graphene oxide films. , 2009, Chemistry.

[23]  Jun Liu,et al.  Glucose oxidase-graphene-chitosan modified electrode for direct electrochemistry and glucose sensing. , 2009, Biosensors & bioelectronics.

[24]  S. Shleev,et al.  Direct heterogeneous electron transfer reactions of fungal laccases at bare and thiol-modified gold electrodes , 2006 .

[25]  X. Xia,et al.  The Electrochemical Properties of 1-Pyrenebutyric acid/Graphene Composites and Their Application in Glucose Biosensors , 2012, Journal of Electrochemistry.

[26]  Juan Tang,et al.  Laccase on Black Pearl 2000 modified glassy carbon electrode: Characterization of direct electron transfer and biological sensing properties for pyrocatechol , 2012 .

[27]  Sergey Shleev,et al.  Direct electron transfer between copper-containing proteins and electrodes. , 2005, Biosensors & bioelectronics.

[28]  N. Durán,et al.  Effects of fungal laccase immobilization procedures for the development of a biosensor for phenol compounds. , 2001, Talanta.

[29]  L. Avigliano,et al.  Unmediated heterogeneous electron transfer reaction of ascorbate oxidase and laccase at a gold electrode. , 1998, The Biochemical journal.

[30]  Bin Wang,et al.  A novel hydrogen peroxide sensor based on the direct electron transfer of horseradish peroxidase immobilized on silica-hydroxyapatite hybrid film. , 2009, Biosensors & bioelectronics.

[31]  De Quan,et al.  Characterization of an amperometric laccase electrode covalently immobilized on platinum surface , 2004 .

[32]  Shen-ming Chen,et al.  A simple electrochemical approach to fabricate a glucose biosensor based on graphene-glucose oxidase biocomposite. , 2013, Biosensors & bioelectronics.

[33]  Hui-Ming Cheng,et al.  Direct reduction of graphene oxide films into highly conductive and flexible graphene films by hydrohalic acids , 2010 .

[34]  Jia Liu,et al.  Immobilization of laccase onto 1-aminopyrene functionalized carbon nanotubes and their electrocatalytic activity for oxygen reduction , 2010 .

[35]  Huangxian Ju,et al.  Reagentless glucose biosensor based on direct electron transfer of glucose oxidase immobilized on colloidal gold modified carbon paste electrode. , 2003, Biosensors & bioelectronics.