An amperometric Meldola Blue-mediated sensor high sensitive to hydrogen peroxide based on immobilization of horseradish peroxidase in a composite membrane of regenerated silk fibroin and poly(vinyl alcohol) (original) (raw)
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Fresenius Journal of Analytical Chemistry, 1997
Horseradish peroxidase (HRP) was effectively entrapped in a novel composite membrane of poly-vinyl alcohol and regenerated silk fibroin, and IR was employed to provide a useful insight into the structure of the composite membrane. A methylene bluemediated sensor highly sensitive to hydrogen peroxide was constructed, which was based on the immobilization of HRP in the composite membrane. Cyclic voltammetry and amperometric measurement were utilized to demonstrate the feasibility of electron communication between immobilized HRP and a glassy carbon electrode in the bioelectrocatalytic reduction of hydrogen peroxide via methylene blue. Performance and characteristics of the sensor were evaluated with regard to response time, detection limit, selectivity, and dependence on temperature and pH as well as operating and storage stability. The sensor possesses a variety of characteristics including high sensitivity, rapid response time and a low detection limit of 0.1 amol/L.
Enzyme and Microbial Technology, 1997
Horseradish peroxidase (HRP) was effectively entrapped in a novel composite membrane of polyvinyl alcohol (PVA) and regenerated silk jibroin (RSF). IR and scanning electron microscopy were employed to provide a useful insight into the structure and morphology of the immobilized HRP composite membrane. A new methylene blue-mediating sensor for hydrogen peroxide was constructed which was based on the composite membrane as an immobilization matrix for HRP. Cyclic voltammetry and amperometric measurements were used to demonstrate the efticiency of the new methylene blue mediating an electron transfer between the immobilized HRP and a glassy carbon electrode in bioelectrocatalytic reduction of hydrogen peroxide. Performance and characteristics of the sensor were evaluated with regard to response time, detection limit, selectivity, and dependences on temperature and pH as well as operating and storage stabilities. The sensor has a variety of characteristics including good sensitivity, rapid response time, and low detection limit of 5.0 PM. 0 1997 Elsevier Science Inc.
Talanta, 1996
Regenerated silk fibroin prepared from waste silk was employed as immobilization matrix for peroxidase and the structures of the blend membranes of regenerated silk fibroin and peroxidase were first investigated with IR and scanning electron microscopy. There was intermolecular interaction between peroxidase and regenerated silk fibroin in the immiscible state. Cyclic voltammetry and constant applied potential measurement showed that Methylene Green efficiently mediated electron transfer from oxidized horseradish peroxidase in regenerated silk fibroin membrane to a glassy carbon electrode. A sensor coupling immobilized peroxidase with Methylene Green responded rapidly to low H,O, concentration and achieved 95% of the steady-state current in less than 25 s with a detection limit of 1.0 x lop7 M H,Oz. The sensor was stable in continuous operation, indicating that peroxidase was entrapped in regenerated silk fibroin membrane and did not freely diffuse away from the sensor surface into solutions.
Biosensors and Bioelectronics, 1997
Horseradish peroxidase (HRP) was immobilized onto a membrane of the regenerated silk fibroin (RSF) from waste silk. The structure of the blend membrane of RSF and HRP was characterized by the use of IR spectra. A second generation of HzO2 sensor on the basis of the immobilized HRP was fabricated, in which tetrathiafulvalene acts as mediating electron transfer between the immobilized enzyme and a glassy carbon electrode. Dependencies of pH and temperature on the H202 biosensor were checked by utilizing cyclic voltammetry. The sensor exhibits high sensitivity, good reproducibility and storage stability.
Electrochimica Acta, 1996
Peroxidase was immobilized in the regenerated silk fibroin membrane prepared from waste silk and the structures of the blend membranes of regenerated silk fibroin and peroxidase were investigated with ir and scanning electron microscopy, indicating that the two kinds of macromolecules were immiscible and their intermolecular interactions existed in the blend membrane. Cyclic voltammetry and constant applied potential measurements tested the catalytic activity of the immobilized peroxidase in water-iso-propyl alcohol system by employing ferrocyanide as an electron transfer mediator between the enzyme and a glassy carbon electrode. Regenerated silk fibroin coating appeared very attractive for entrapping the enzyme onto the electrode since the immobilized peroxidase showed long-term stability in the organic media.
Enzyme Engineering, 2016
An enzymatic biosensor has been developed for detection of hydrogen peroxide with immobilized Horseradish peroxidase (HRP). HRP was immobilized by using glutaraldehyde (GA) that cross linked with modified polyaniline (PANI) as mediator to improve electron transfer. Modified carbon paste electrodes (MCPE) PANI was more effective during electron transfer compared to carbon paste electrodes (CPE). Cyclic voltammetry method (VC) was used to determine the electrochemical properties of the modified electrode substrate to produce redox reactions. The effect of pH and temperature were analyzed by cyclic voltammetry. The optimum performance of HRP/GA/PANI was at pH 7 and 50°C. Kinetics parameters HRP enzyme were determined in optimum condition. The Michaelis-Menten constant (Km) value and current maximum (I max) have been obtained as 1.71 mM and 0.29 mA.
Sensors, 2008
A modified electrode for hydrogen peroxide (H 2 O 2) sensing was prepared via thiophene (Th) with epoxy group. Thiophene (EpoxyTh) with epoxy group was synthesized by reaction of 3-bromothiophene and glycidyl methacrylate (GMA) in acetonitrile according to Heck Reaction. The electrocopolymerization of Th and EpoxyTh was performed on the surface of indium tin oxide (ITO) electrode by cycling the potential between-1.0 and +2.5 V in mixture of thiophene (Th) and EpoxyTh. Poly(Th-co-EpoxyTh) grown onto the ITO electrode was successfully confirmed by SEM, AFM, and water contact angle analysis, respectively. Finally, the HRP was immobilized on the surface of poly(Th-co-EpoxyTh) electrode by covalent binding. The amperometric response of the HRP-immobilized poly(Th-co-EpoxyTh) electrode for H 2 O 2 was examined by cyclic voltammetry (CV). The HRP-immobilized poly(Th-co-EpoxyTh) electrode showed linearity from 0.1 to 30 mM H 2 O 2 , good reproducibility, and long life time.
Horse radish peroxidase immobilized polyaniline for hydrogen peroxide sensor
Polymers for Advanced Technologies, 2009
Horse radish peroxidase (HRP) has been electrochemically entrapped into perchlorate (ClO S 4) doped polyaniline (PANI) film deposited onto indium-tin-oxide (ITO) coated glass plate. This HRP-PANI-ClO S 4 /ITO bioelectrode characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), UV-Visible spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) techniques has been utilized for estimation of hydrogen peroxide (H 2 O 2). This H 2 O 2 sensor exhibits response time of 5 s, linearity from 3 to 136 mM, sensitivity as 0.5638 mA mM S1 cm S2 with linear regression of 0.985. The value of the Michaelis-Menten constant (K m) has been obtained as 1.984 mM.
Analytica Chimica Acta, 1998
An amperometric enzyme electrode for monitoring hydrogen peroxide (H 2 O 2) was constructed by coimmobilising periodate oxidised horseradish peroxidase (HRP) and a redox polymer containing a covalently bound phenothiazine type mediator, toluidine blue O (TBO), in a carbon paste matrix. Two different carbon paste electrodes were designed using either paraf®n oil or solid paraf®n wax as binders. A new mode of preparing enzyme modi®ed carbon±wax composites is presented based on dissolving the wax in n-pentane before mixing with the enzyme modi®ed carbon powder excluding the necessity of melting the wax prior to mixing. The developed sensors were used in a¯ow-injection system at an applied potential of À50 mV vs. Ag/AgCl. Both electrode con®gurations displayed superior storage and operational stability, compared with similarly prepared electrodes containing only native HRP, where the electrical communication was based on a direct electron transfer between the electrode and HRP. Detection limits (calculated as twice the signal to noise ratio) of 50 nM H 2 O 2 were obtained for these electrodes.
Biosensors and Bioelectronics, 2009
MWCNTs-nanoNiO composite was used as a glassy carbon electrode modifier for construction of a novel catalase nanobiosensor for hydrogen peroxide. The immobilized catalase exhibited excellent electrocatalytic activity towards the reduction of H 2 O 2. The resulting amperometric biosensor exhibited a linear response over a concentration range of 200 mM to 2.53 mM with a low detection limit of 19.0 mM. Electrochemical impedance measurements revealed that the modified electrode can be used for the sensitive detection of H 2 O 2. The charge transfer resistance found to decrease significantly after enzymatic reaction of nanobiosensor with H 2 O 2. The resulting impedance was highly sensitive to H 2 O 2 over a linear range of 19-170 nM with a detection limit of 2.4 nM.