Cyanide detection using a substrate-regenerating, peroxidase-based biosensor.

An enzyme-based, dual working electrode system is described for the sensing of cyanide. Horseradish peroxidase (HRP) is incorporated as the sensing element. A continuous monitoring of oxidative activity by the enzyme results through the generation and regeneration of substrates at the electrode surfaces. Thus, HRP is oxidized by hydrogen peroxide generated from dissolved oxygen, at the primary electrode, and then reduced through the secondary electrode by mediated electron transfer using ferrocene as a carrier. Ferrocene regeneration at this electrode is proportional to the intrinsic activity of HRP. The dynamics of the system are investigated by using a rotating ring-disk electrode. The enzyme is immobilized to provide better control over its catalytic activity and to increase the lifetime of the biosensor. Cyanide inhibition of current can be modeled by reversible binding kinetics. Detection of cyanide is possible in submicromolar (ppb) concentrations, with a half maximal response at 2 microM. The response time for detection of introduced cyanide is within 1 s. The sensor can be operated between 5 and 40 degrees C, and cyanide inhibition is unaffected by pH changes between 5 and 8. The sensor is reproducible for cyanide determination and is stable for over 6 months.

[1]  A. Cass,et al.  Ferricinium ion as an electron acceptor for oxido-reductases , 1985 .

[2]  W. Clark.  CIV.—The electrometric titration of halides , 1926 .

[3]  H. Dunford.  On the function and mechanism of action of peroxidases , 1976 .

[4]  J. F. Alder,et al.  Optical fibre sensor for detection of hydrogen cyanide in air , 1989 .

[5]  P. Ross,et al.  LEED/electrochemical analysis of Au single crystals: Stability of the UHV prepared surfaces of Au(111) and Au(100) in aqueous electrolyte , 1987 .

[6]  S. Motoo,et al.  Effect of terraces and steps in the electrocatalysis for formic acid oxidation on platinum , 1987 .

[7]  R. Behm,et al.  An in-situ scanning tunneling microscopy study of au (111) with atomic scale resolution , 1988 .

[8]  M. J. Cormier,et al.  An investigation of the mechanifm of the luminescent peroxidation of luminol by stopped flow techniques. , 1968, The Journal of biological chemistry.

[9]  P. Hansma,et al.  Scanning tunneling microscopy and atomic force microscopy: application to biology and technology. , 1988, Science.

[10]  A. Turner,et al.  Ferrocene-mediated enzyme electrode for amperometric determination of glucose. , 1984, Analytical chemistry.

[11]  G. S. Wilson,et al.  Rotating ring-disk enzyme electrode for biocatalysis kinetic studies and characterization of the immobilized enzyme layer , 1980 .

[12]  H. Hill,et al.  A method for estimation of hydrogen peroxide based on mediated electron transfer reactions of peroxidases at electrodes , 1986 .

[13]  P. Nicholls.  The site of respiratory inhibition by cyanide , 1983 .

[14]  A. Cass,et al.  Inhibited enzyme electrodes. Part 1: Theoretical model. , 1990, Biosensors & bioelectronics.

[15]  G. Marr,et al.  Oxidation of ferrocene and some substituted ferrocenes in the presence of horseradish peroxidase , 1978 .

[16]  M. Mehra,et al.  Detection and determination of cyanide--a review. , 1986, International journal of environmental analytical chemistry.

[17]  S. Cadle,et al.  A ring-disk study of the effect of trace chloride ion on the anodic behavior of gold in 0.2 M H2SO4 , 1973 .

[18]  W. D. Hewson,et al.  Horseradish peroxidase. XXIX. Reactions in water and deuterium oxide: cyanide binding, compound I formation, and reactions of compounds I and II with ferrocyanide , 1978 .

[19]  P K Hansma,et al.  Atomic-Resolution Microscopy in Water , 1986, Science.

[20]  Kingo Itaya,et al.  In situ scanning tunneling microscopy of platinum (111) surface with the observation of monatomic steps , 1990 .

[21]  Wilson,et al.  Observation of atomic corrugation on Au(111) by scanning tunneling microscopy. , 1987, Physical review letters.

[22]  D. Dolman,et al.  A kinetic study of the reaction of horseradish peroxidase with hydrogen peroxide. , 1975, Canadian journal of biochemistry.

[23]  Albericci Vj.  Rapid colorimetric estimation of iodine in kelp. , 1945 .

[24]  B. Wu,et al.  Electrochemical study of the initial surface condition of platinum surfaces with (100) and (111) orientations , 1982 .

[25]  C. Chidsey,et al.  In situ scanning-tunneling-microscope observation of roughening, annealing, and dissolution of gold (111) in an electrochemical cell , 1989 .

[26]  Paul K. Hansma,et al.  Tunneling microscopy, lithography, and surface diffusion on an easily prepared, atomically flat gold surface , 1988 .

[27]  Ronald Woods,et al.  A study of the dissolution of platinum, palladium, rhodium and gold electrodes in 1 m sulphuric acid by cyclic voltammetry , 1972 .

[28]  N. Matsuda,et al.  Surface electronic structure of semiconductor ( p‐ and n‐Si) electrodes in electrolyte solution , 1990 .

[29]  Z. Shu,et al.  Direct electrochemical studies of ligand-binding reactions to cytochrome c oxidase , 1986 .

[30]  H. Angerstein-Kozlowska,et al.  Elementary steps of electrochemical oxidation of single-crystal planes of Au Part II. A chemical and structural basis of oxidation of the (111) plane , 1987 .

[31]  A. Bard,et al.  In situ scanning tunneling microscopy of the anodic oxidation of highly oriented pyrolytic graphite surfaces , 1988 .

[32]  H. Hill,et al.  Enzyme dual-electrode for analyte determination , 1989 .

[33]  H. Girault,et al.  Interdigitated microband electrodes: chronoamperometry and steady state currents , 1989 .

[34]  D. Keilin,et al.  On the Haematin Compound of Peroxidase , 1937 .

[35]  R. Carr,et al.  In situ scanning tunneling microscopy studies of the underpotential deposition of lead on Au(111) , 1989 .

[36]  S. Cadle,et al.  Ring-disk electrode study of the anodic behavior of gold in 0.2M sulfuric acid , 1974 .

[37]  P. Tijssen,et al.  Highly efficient and simple methods for the preparation of peroxidase and active peroxidase-antibody conjugates for enzyme immunoassays. , 1984, Analytical biochemistry.

[38]  G. S. Wilson,et al.  Rotating ring-disk enzyme electrode for surface catalysis studies. , 1976, Analytical chemistry.

[39]  P. Hansma,et al.  Semiconductor topography in aqueous environments: tunneling microscopy of chemomechanically polished (001) GaAs , 1987 .

[40]  H. Dunford,et al.  Studies on horseradish peroxidase. 13. The kinetic effect of cyanide on the oxidation-reduction cycle. , 1973, Canadian journal of biochemistry.

[41]  K. Itaya,et al.  Scanning tunneling microscope for electrochemistry ― a new concept for the in situ scanning tunneling microscope in electrolyte solutions , 1988 .

[42]  K. Itaya,et al.  In situ scanning tunneling microscopy for platinum surfaces in aqueous solutions , 1988 .

[43]  A. Hubbard.  Electrochemistry at well-characterized surfaces , 1988 .

[44]  J. Moiroux,et al.  Enzymic electrocatalysis: electrochemical regeneration of NAD+ with immobilized lactate dehydrogenase modified electrodes , 1984 .

[45]  Jaklevic,et al.  Scanning-tunneling-microscope observation of surface diffusion on an atomic scale: Au on Au(111). , 1988, Physical review letters.

[46]  D. Kolb.  UHV Techniques in the Study of Electrode Surfaces , 1987 .

[47]  H. Dunford,et al.  Studies on Horseradish Peroxidase. XI. On the Nature of Compounds I and II as Determined from the Kinetics of the Oxidation of Ferrocyanide , 1973 .

[48]  S. Motoo,et al.  Hydrogen and oxygen adsorption on Ir (111), (100) and (110) planes , 1984 .