Novel enzyme biosensor for hydrogen peroxide via supramolecular associations (original) (raw)
Related papers
Journal of Electroanalytical Chemistry, 2000
A novel immobilization approach based on the supramolecular function between b-cyclodextrin polymer (b-CDP) and cationic dyes, and the condensation polymerization among b-CDP, glutaric dialdehyde and horseradish peroxidase (HRP) in the fabrication of a hydrogen peroxide sensor is described. IR and UV-vis spectroscopy were employed to characterize the structure of the composite membrane modified on the surface of the electrode. AFM was used to visualize the morphology of the composite membrane. The characteristics of supramolecular inclusion compounds between b-CDP and cationic dyes were studied. The fabricated H 2 O 2 sensor responded rapidly to H 2 O 2 in the linear range from 1.0 to 1.1 mmol l − 1 with a detection limit of 0.5 mol l − 1 .
Solid State Ionics, 2018
A comparison was made between two plant peroxidases, cationic horseradish peroxidase (HRP) and anionic tobacco peroxidase (TOP), combined with a highly cationic osmium polymer [Os(4,4′-dimethyl-2,2′-bipyridine) 2 poly(N-vinylimidazole) 10 Cl] + 2/+ ([Os(dmp)PVI] +/2 +) to develop highly sensitive, stable and selective hydrogen peroxide biosensors. The two different plant peroxidases were individually immobilized onto graphite rod (G) electrodes by a three steps drop-casting procedure consisting of the subsequent deposition of an aqueous solution of ([Os(dmp)PVI] +/2 +), followed by a solution of poly(ethyleneglycol) diglycidyl ether (PEGDGE), used as a cross linking agent and finally an aliquot of a solution of cationic HRP or anionic TOP to make HRP/ PEGDGE/[Os(dmp)PVI] +/2 + /G and TOP/PEGDGE/[Os(dmp)PVI] +/2 + /G based electrodes, respectively. Electrochemical experiments were carried out to investigate the influence of the surface charge of the enzyme and the charge of the polymer on the efficiency of the electron transfer (ET) between the enzyme and the wiring redox polymer and the efficiency for electrocatalytic reduction of H 2 O 2. In the case of HRP a decrease in the ET rate was observed due to the repulsion between this enzyme and the polymer, both positively charged, whereas with TOP there was an enhanced ET rate due to the attraction between the anionic enzyme and the cationic polymer. The effects of enzyme loading and pH were investigated. Both peroxidase modified electrodes exhibited a wide dynamic response range (1-500 μM H 2 O 2) and a low detection limit (0.3 μM H 2 O 2). The TOP based electrode showed a higher sensitivity (470 nA μM − 1 cm − 2) compared to that of the HRP based electrode (300 nA μM − 1 cm − 2) and an improved long-term stability (decrease in 17.3% upon 30 days compared with 50% for HRP). Both enzyme electrodes showed a response time of 3 s. The HRP based sensor was more sensitive to the presence of phenolic compounds acting as alternative electron donors, whereas the TOP based sensor was virtually interference free. Both HRP and TOP based electrodes were successfully tested in contact lens cleaning samples and real "spiked" samples from different sources such as tap water, milk and dairy products.
Electrochimica Acta, 2011
Gold electrodes were functionalized with an electropolymerized matrix of Au nanoparticles modified with 2-mercaptoethanesulfonic acid, 3-mercaptophenyl boronic acid and p-aminothiophenol. The resulting nanostructured electroconductive matrix was used as support for the oriented immobilization of horseradish peroxidase to construct a reagentless amperometric biosensor for H 2 O 2. The electrode, poised at 0.0 mV, exhibited a rapid response within 8 s and a linear calibration range from 5 M to 1.1 mM H 2 O 2. The sensitivity of the biosensor was determined as 498 A/M cm 2 , and its detection limit was 1.5 M H 2 O 2 at a signal-to-noise ratio of 3. The electrode retained 95% and 72% of its initial activity after 21 and 40 days of storage at 4 • C.
Sensors and Actuators B: Chemical, 2010
In this work, a new type of amperometric hydrogen peroxide biosensor was fabricated for the determination of H2O2. Horseradish peroxidase (HRP) was immobilized on a glassy carbon electrode by poly(glycidyl methacrylate-co-vinylferrocene) (poly(GMA-co-VFc)) film. A polymeric electron transfer mediator, containing copolymers of glycidyl methacrylate (GMA) and vinylferrocene (VFc) with different molar ratios, was prepared by free-radical copolymerization. The amperometric response was measured as a function of H2O2 concentration, at a fixed potential of +0.35 V vs. Ag/AgCl in phosphate-buffered saline (pH 7.0). The mediated hydrogen peroxide biosensor showed a fast response of less than 4 s of linear range 2.0–30.0 mM, with a detection of 2.6 μM. The sensitivity of the biosensor for H2O2 was 10.42 nA/mM cm2.
Electroanalysis, 2008
A novel method to fabricate a third-generation hydrogen peroxide biosensor was reported. The electrode was first derivatized by electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt (4-CPDS) in acidic aqueous solution yielded stable 4-carboxyphenyl (4-CP) layer. The horseradish peroxidase (HRP) enzyme was then covalently immobilized by amidation between NH2 terminus of enzyme and COOH terminus of 4-CP film making use of the carbodiimide chemistry. Electrodeposition conditions used to control electrode functionalization density and film electron transfer kinetics were assessed by chronoamperometry and electrochemical impedance spectroscopy. The immobilized HRP displayed excellent electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2) without any mediators. The effect of various operational parameters was explored for optimum analytical performance. The reported biosensor exhibited fast amperometric response (within 5 s) to H2O2. The detection limit of the biosensor was 5 μM, and linear range was from 20 μM to 20 mM. Furthermore, the biosensor exhibited high sensitivity, good reproducibility, and long-term stability.
Journal of Electroanalytical Chemistry, 1998
A new approach to construct a reagentless hydrogen peroxide biosensor is described. Horseradish peroxidase and thionine are covalently bound to a cysteamine-assembled gold electrode using glutaraldehyde as a bifunctional reagent. Thionine immobilised in this way can shuttle electrons between the electrode and the redox activity center of the enzyme. The sensor was highly sensitive to hydrogen peroxide with a detection limit of 8.0× 10 − 7 mol l − 1 and a response time of less than 4 s. The effects of the applied potential and the pH values of the buffer solution on the response of the sensor were investigated for optimum analytical performance.
Analytical Chemistry, 1999
Exposure of gold surfaces to solutions of dithiobis Nsuccinimidyl propionate (DTSP) gives rise to the modification of the surface with N-succinimidyl-3-thiopropionate (NSTP) which can, in turn, react with amino groups allowing for the covalent immobilization of enzymes such as horseradish peroxidase (HRP). The coverage of NSTP has been estimated to be of the order of 1.3 × 10-10 from the charge consumed during its reductive desorption. The binding reaction of HRP with NSTP modified gold surfaces has been studied with the quartz crystal microbalance, and the results suggest that the immobilization process involves two steps in which the first (faster) appears to correspond to the rapid incorporation of the enzyme whereas the second is likely due to the slow incorporation of additional enzyme and/or reorganization of the immobilized layer. Spectrophotometric and electrochemical assays indicate that the immobilized HRP retains its enzymatic activity after immobilization onto the DTSP modified gold surface. The amount of immobilized (and active) HRP was estimated from QCM and spectrophotometric measurements to be of the order of 1.5 × 10-11 mol/cm 2. A peroxide biosensor was developed making use of a gold surface modified with DTSP and HRP employing Os and Ru complexes of 1,10-phenanthroline 5,6-dione (phen-dione) of the type [M(phendione) x (L) 3-x ] +2 (where L) 1,10-phenanthroline or 2,2′bipyridine, x) 1-3) as mediators with the quinone moieties being the active component. The efficiency of the mediators increased with increasing number of phendione ligands.
Korean Journal of Chemical Engineering, 2012
An electrochemical hydrogen peroxide biosensor was designed by immobilizing horseradish peroxidase (HRP) on Ag nanoparticles/cysteamine/p-aminobenzene sulfonic acid/glassy carbon (GC) electrode. Ag nanoparticles can act as tiny conduction centers on electrodes that adsorb redox enzymes, facilitating the transfer of electrons with no requiring any loss of biological activity. The forerunner film was first electropolymerized on the glassy carbon electrode with p-aminobenzene sulfonic acid (p-ABSA) by cyclic voltammetry. The cysteamine (CA) was bound on the surface of the film by electrostatic force, then Ag nanoparticles were immobilized on the cysteamine monolayer, and lastly HRP was adsorbed onto the surfaces of the Ag nanoparticles. A dramatic decrease in the overvoltage of H 2 O 2 was observed with improved sensitivity, which makes the modified electrodes of great promise for oxidase-based amperometric biosensors. The biosensor responded to H 2 O 2 in the linear range from 1.2×10 6 mol/L to 9.8×10 3 mol/L with a detection limit of 1.1×10 8 mol/L. Moreover, the obtained biosensor exhibited good accuracy and high sensitivity.
A poly(o-aminophenol) modified electrode as an amperometric hydrogen peroxide biosensor
Electrochimica Acta, 1998
AbstractÐAn enzymatic biosensor for hydrogen peroxide based on a horseradish peroxidase (HRP)-ferrocene carbon paste modi®ed electrode and coated with a layer of electrochemically generated poly(o-aminophenol) is reported. A linear calibration curve is obtained over the range 1 Â 10 À8 M to 1 Â 10 À5 M. The biosensor responds to hydrogen peroxide in a few seconds and has a detection limit of 8.5 Â 10 À9 M. The response of the biosensor is diusion controlled at low substrate concentrations. Flow injection assays of hydrogen peroxide at a sampling rate of 150 injections per hour with a relative standard deviation of 0.8% (50 samples) are possible. Applicability of the sensor for measurement of hydrogen peroxide in real samples (milk) was demonstrated. #