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1.
《Electroanalysis》2017,29(12):2698-2707
A cholesterol biosensor based on cholesterol oxidase‐poly(diallyldimethylammonium chloride)‐carbon nanotubes‐nickel ferrite nanoparticles (ChOx‐PDDA‐CNTs‐NiFe2O4NPs) solution is easily fabricated by using a single dropping step on a glassy carbon electrode (GCE) surface. This technique is an alternative way to reduce complexity, cost and time to produce the biosensor. The uniformly dispersed materials on the electrode surface enhance the catalytic reaction of cholesterol oxidase and electron transfer from the oxidation of hydrogen peroxide in the system. The nickel ferrite nanoparticles were synthesized by co‐precipitation and calcination at various temperatures. These nanoparticles were then characterized using field emission scanning electron microscopy (FE‐SEM), energy‐dispersive X‐ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), cyclic voltammetry (CV) and X‐ray diffraction (XRD). The synthesized material calcined at 700 °C was well defined and presented the octahedral metal stretching with cubic NiFe2O4NPs phase. In cyclic voltammetric study, the ChOx‐PDDA‐CNTs‐NiFe2O4NPs/GCE showed 0.43 s−1 charge transfer rate constant (K s), 7.79×10−6 cm2 s−1 diffusion coefficient value (D ), 0.13 mm2 electroactive surface area (A e) and 3.58×10−8 mol cm−2 surface concentration ( ). This modified electrode exhibits stability in term of percent relative standard deviation (%RSD=0.62 %, n=10), reproducibility (%RSD=0.81, n=10), high sensitivity (25.76 nA per mg L−1 cm−2), linearity from 1 to 5,000 mg L−1 (R2=0.998) with a low detection limit (0.50 mg L−1). Its Michaelis‐Menten constant (K m) was 0.14 mM with 0.92 μA maximum current (I max) and demonstrated good selectivity without the effects of electroactive species such as ascorbic acid, glucose and uric acid. The cholesterol biosensor was successfully applied to determine cholesterol levels in human blood samples, showing promise due to its simplicity and availability.  相似文献   

2.
A novel composite material of copper (I) oxide at manganese (IV) oxide (Cu2O@MnO2), was synthesized and applied for modification on the glassy carbon electrode (GCE) surface (Cu2O@MnO2/GCE) as a hydrogen peroxide (H2O2) sensor. The composite material was characterized regarding its structural and morphological properties, using field emission scanning electron microscopy (FE‐SEM), energy‐dispersive X‐ray spectroscopy (EDX), X‐ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The Cu2O@MnO2/GCE showed an excellent electrocatalytic response to the oxidation of H2O2 which provided a 0.56 s?1 charge transfer rate constant (Ks), 1.65×10?5 cm2 s?1 diffusion coefficient value (D), 0.12 mm2 electroactive surface area (Ae) and 1.04×10?8 mol cm?2 surface concentration ( ). At the optimal condition, the constructed sensor exhibited a wide linear range from 0.5 μM to 20 mM with a low limit of detection (63 nM, (S/N=3) and a good sensitivity of 256.33 μA mM?1 cm?2. It also presented high stability (ΔIresponse±15 %, n=100), repeatability (1.25 %RSD, n=10) and reproducibility (3.55 %RSD, n=10). The results indicated that the synthesized Cu2O@MnO2 was successfully used as a new platform for H2O2 sensing.  相似文献   

3.
在碳纳米管(CNTs)和K3Fe(CN)6修饰的铂电极上吸附固定胆碱氧化酶,以鲁米诺为发光试剂,研制了胆碱电化学发光(ECL)生物传感器.CNTs可有效提高电极表面的电荷传输能力、提高电极表面的生物相容性和对酶分子的固载能力;K3Fe(CN)6对酶活性具有激活作用,同时对H2O2增敏的鲁米诺ECL有增强作用,均有利于提...  相似文献   

4.
A simple, selective and stable biosensor with the enzymatic reactor based on choline oxidase (ChOx) was developed and applied for the determination of choline (Ch) in flow injection analysis with amperometric detection. The enzyme ChOx was covalently immobilized with glutaraldehyde to mesoporous silica powder (SBA‐15) previously covered by NH2‐groups. This powder was found as an optimal filling of the reactor. The detection of Ch is based on amperometric monitoring of consumed oxygen during the enzymatic reaction, which is directly proportional to Ch concentration. Two arrangements of an electrolytic cell in FIA, namely wall‐jet cell with working silver solid amalgam electrode covered by mercury film and flow‐through cell with tubular detector of polished silver solid amalgam were compared. The experimental parameters affecting the sensitivity and stability of the biosensor (i. e. pH of the carrier solution, volume of reactor, amount of the immobilized enzyme, the detection potential, flow rate, etc.) were optimized. Under the optimized conditions, the limit of detection was found to be 9.0×10?6 mol L?1. The Michaelis‐Menten constant for covalently immobilized ChOx on SBA‐15 was calculated. The proposed amperometric biosensor with the developed ChOx‐based reactor exhibits good repeatability, reproducibility, long‐term stability, and reusability. Its efficiency has been confirmed by the successful application for the determination of Ch in two commercial pharmaceuticals.  相似文献   

5.
An effective, stable enzymatic glucose biosensor was fabricated on a glassy carbon electrode (GCE) surface using simple multicomposite materials (MCM): a solution of prepared poly(diallyldimethylammonium chloride)‐capped gold nanoparticles‐nickel ferrite particles‐carbon nanotubes‐chitosan (PDDA‐AuNPs‐NiFe2O4‐CNTs‐CHIT), electropolymerization of poly(o‐phenylenediamine) (PoPD) and immobilization of glucose oxidase (GOx). Biocompatibility and synergy of the MCM enhanced the immobilization and the reaction of GOx and as well as the electron transfer from an oxidation reaction of hydrogen peroxide in the system. The NiFe2O4 was synthesized by co‐precipitation and calcined at 700 °C. Characterization was carried out by field emission scanning electron microscopy (FE‐SEM), energy‐dispersive X‐ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR) and X‐ray diffraction (XRD) which presented both tetrahedral and octahedral metal stretching with a cubic NiFe2O4 crystal phase. The GOx/PoPD/MCM/GCE yielded a 0.77 s?1 charge transfer rate constant (Ks), a 2.28×10?6 cm2 s?1 diffusion coefficient value (D), a 0.21 mm2 electroactive surface area (Ae) and a 1.93×10?8 mol cm?2 surface concentration ( ) as determined by cyclic voltammetry. The modified electrode showed a durable operation time (n=97, more than 50 % I), repeatability (%RSD=0.38, n=10), reproducibility (%RSD=1.60, n=10), high sensitivity (853.07 μA mM?1 cm?2), selectivity without effects of electroactive species (aspirin, uric acid, caffeine, cholesterol, ascorbic acid and dopamine) and two linear ranges from 0.5 to 10 μM (R2=0.998) and 10 to 15,000 μM (R2=0.991) with a low detection limit (0.35 μM, S/N=3). Its Michaelis‐Menten constant (Km) was calculated as 93.51 μM with 46.30 μA maximum current (Imax). This proposed simple method was successfully applied for glucose determination in human blood samples.  相似文献   

6.
A microelectrode array microprobe with a choline sensing site and an on‐probe reference electrode was constructed by depositing permselective polymer films and choline oxidase (ChOx) on one microelectrode, and iridium oxide (IrOx) on another, both of which were coated previously with a nanostructured Pt deposit. Scanning electron microscopy (SEM) of the nanostructured Pt layer revealed a unique pillar‐like, “nanograss” structure. Polyphenylenediamine (PPD) and Nafion were coated sequentially on the working (i. e. sensing) electrode surface to serve as the permselective polymer films. The microsensor exhibited high sensitivity to choline (123±13 μA mM?1 cm?2), low detection limit (3.2±0.8 μM), and fast response time (3–5 s). The choline sensor also was tested at physiological concentrations of electroactive interfering species common to brain extracellular fluid (i. e. ascorbic acid, dopamine, DOPA, and DOPAC) and showed excellent selectivity. Selectivity likely was aided by the relatively low potential of 0.35 V vs. IrOx that was made possible by the enhanced H2O2 electrooxidation activity of the underlying nanostructured Pt‐coated working electrode. Thus, Pt “nanograss” appears to be an excellent electrode surface modification for creation of high performance electroenzymatic biosensors.  相似文献   

7.
Shi H  Yang Y  Huang J  Zhao Z  Xu X  Anzai J  Osa T  Chen Q 《Talanta》2006,70(4):852-858
An amperometric choline biosensor was developed by immobilizing choline oxidase (ChOx) in a layer-by-layer (LBL) multilayer film on a platinum (Pt) electrode modified with Prussian blue (PB). 6-O-Ethoxytrimethylammoniochitosan chloride (EACC) was used to prepare the ChOx LBL films. The choline biosensor was used at 0.0 V versus Ag/AgCl to detect choline and exhibited good characteristics such as relative low detection limit (5 × 10−7 M), short response time (within 10 s), high sensitivity (88.6 μA mM−1 cm−2) and a good selectivity. The results were explained based on the ultrathin nature of the LBL films and the low operating potential that could be due to the efficient catalytic reduction of H2O2 by PB. In addition, the effects of pH, temperature and applied potential on the amperometric response of choline biosensor were evaluated. The apparent Michaelis-Menten constant was found to be (0.083 ± 0.001) ×10−3 M. The biosensor showed excellent long-term storage stability, which originates from a strong adsorption of ChOx in the EACC multilayer film. When the present choline biosensor was applied to the analysis of phosphatidylcholine in serum samples, the measurement values agreed satisfactorily with those by a hospital method.  相似文献   

8.
《Electroanalysis》2017,29(7):1741-1748
The determination of lead ions by inhibition of choline oxidase enzyme has been evaluated for the first time using an amperometric choline biosensor. Choline oxidase (ChOx) was immobilized on a glassy carbon electrode (GCE) modified with multiwalled carbon nanotubes (MWCNT) through cross‐linking with glutaraldehyde. In the presence of ChOx, choline was enzymatically oxidized into betaine at –0.3 V versus Ag/AgCl reference electrode, lead ion inhibition of enzyme activity causing a decrease in the choline oxidation current. The experimental conditions were optimised regarding applied potential, buffer pH, enzyme and substrate concentration and incubation time. Under the best conditions for measurement of the lowest concentrations of lead ions, the ChOx/MWCNT/GCE gave a linear response from 0.1 to 1.0 nM Pb2+ and a detection limit of 0.04 nM. The inhibition of ChOx by lead ions was also studied by electrochemical impedance spectroscopy, but had a narrower linear response range and low sensitivity. The inhibition biosensor exhibited high selectivity towards lead ions and was successfully applied to their determination in tap water samples.  相似文献   

9.
Direct electrochemistry of cholesterol oxidase (ChOx) immobilized on the conductive poly‐3′,4′‐diamine‐2,2′,5′,2″‐terthiophene (PDATT) was achieved and used to create a cholesterol biosensor. A well‐defined redox peak was observed, corresponding to the direct electron transfer of the FAD/FADH2 of ChOx, and the rate constant (ks) was determined to be 0.75 s?1. Glutathione (GSH) covalently bonded with PDATT was used as a matrix for conjugating AuNPs, ChOx, and MP, simultaneously. MP co‐immobilized with ChOx on the AuNPs‐GSH/PDATT exhibited an excellent amperometric response to cholesterol. The dynamic range was from 10 to 130 μM with a detection limit of 0.3±0.04 μM.  相似文献   

10.
In this research, poly(diallyldimethylammonium chloride)-capped gold nanoparticles, nickel ferrite particles, and carbon nanotubes were combined to form a PANC metal composite. The prepared metal composite modified onto a glassy carbon electrode was electropolymerized with poly(o-phenylenediamine) and immobilized with horseradish peroxidase, anti-carcinoembryonic antigen antibody, and bovine serum albumin to create the label-free immunosensors for rapid detection of carcinoembryonic antigen (CEA) using chronoamperometry. This developed biocomposite material modified onto a glassy carbon electrode presented an excellent electrocatalytic response to the redox reaction of hydrogen peroxide as a sensing probe, from which the kinetic parameters including of a charge transfer rate constant, a diffusion coefficient value, an electroactive surface area, and a surface concentration were calculated to be 1.85 s−1, 4.28×10−6 cm2 s−1, 0.14 cm2 and 1.87×10−8 mol cm−2, respectively. The developed immunosensors also exhibited a wide linear range of CEA concentration from 0.01 to 25 ng mL−1 with high sensitivity (96.21 μA cm−2 ng−1 mL) and low detection limit (0.72 pg mL−1), excellent selectivity without interfering effects from possible species (amoxicillin, ascorbic acid, aspirin, caffeine, cholesterol, dopamine, glucose, and uric acid), outstanding stability (n=100, %I>50 %), repeatability (%RSD=0.34, n=10), reproducibility (%RSD=4.06, n=10), and rapid analysis (25 s each operation time). This proposed method was successfully applied for CEA detection in whole blood samples with satisfactory results, suggesting that this developed sensing platform may be considered to be exploited for fabrication of other label-free electrochemical immunosensors for the real sample analysis.  相似文献   

11.
An interesting mode of reactivity of MnO2 nanoparticles modified electrode in the presence of H2O2 is reported. The MnO2 nanoparticles modified electrodes show a bi-direction electrocatalytic ability toward the reduction/oxidation of H2O2. Based on this property, a choline biosensor was fabricated via a direct and facile electrochemical deposition of a biocomposite that was made of chitosan hydrogel, choline oxidase (ChOx) and MnO2 nanoparticles onto a glassy carbon (GC) electrode. The biocomposite is homogeneous and easily prepared and provides a shelter for the enzyme to retain its bioactivity. The results of square wave voltammetry showed that the electrocatalytic reduction currents increased linearly with the increase of choline chloride concentration in the range of 1.0 × 10−5 –2.1 × 10−3 M and no obvious interference from ascorbic acid and uric acid was observed. Good reproducibility and stability were obtained. A possible reaction mechanism was proposed.  相似文献   

12.
This paper reports on the electrochemical behavior of an ATP biosensor that utilizes glucose oxidase (GOx) and hexokinase (Hex) immobilized within the electroactive polymer, polyethylenedioxythiophene (PEDOT). This biosensor design detects ATP indirectly at 0.85 V vs. Ag/AgCl based on the oxidation current for enzymatically generated H2O2, and at −0.20 V; a potential at which improved analyte selectivity is achieved. The detection figures of merit at both detection potentials are a response time of 15±1 s, an experimental detection limit of 10.0±0.2 μmol L−1 (S/N=3), and a sensitivity in the range of 100–500 mA M−1cm−2.  相似文献   

13.
Direct electron transfer of immobilized superoxide dismutase (Cu, Zn‐SOD) onto silicon carbide (SiC) nanoparticles displays a pair of well defined and nearly reversible redox peaks with formal potential (′) of −0.03 V in pH 7.4. The heterogeneous electron transfer rate constant (ks) and surface coverage (Γ) of immobilized SOD are 11.0±0.4 s−1 and 1.42×10−11 mol cm−2. Biosensor shows fast amperometric response (3s) with sensitivity and detection limit of 1.416 nA μM−1, 1.66 μM, and 1.375 nA μM−1, 2.1 μM for cathodically or anodically detection of superoxide, respectively. This biosensor also exhibits good stability, reproducibility and long life‐time.  相似文献   

14.
Cavity ring‐down UV absorption spectroscopy was used to study the kinetics of the recombination reaction of FCO radicals and the reactions with O2 and NO in 4.0–15.5 Torr total pressure of N2 diluent at 295 K. k(FCO + FCO) is (1.8 ± 0.3) × 10−11 cm3 molecule−1 s−1. The pressure dependence of the reactions with O2 and NO in air at 295 K is described using a broadening factor of Fc = 0.6 and the following low (k0) and high (k) pressure limit rate constants: k0(FCO + O2) = (8.6 ± 0.4) × 10−31 cm6 molecule−1 s−1, k(FCO + O2) = (1.2 ± 0.2) × 10−12 cm3 molecule−1 s−1, k0(FCO + NO) = (2.4 ± 0.2) × 10−30 cm6 molecule−1 s−1, and k (FCO + NO) = (1.0 ± 0.2) × 10−12 cm3 molecule−1 s−1. The uncertainties are two standard deviations. © 2001 John Wiley & Sons, Inc. Int J Chem Kinet 33: 130–135, 2001  相似文献   

15.
Electrodeposited cobalt oxide (CoOx) nanomaterials are not only used for immobilization of cholesterol oxidase (ChOx) but also as electron transfer mediator for oxidation of H2O2 generated in the enzymatic reaction. Voltammetry and flow injection analysis (FIA) were used for determination of cholesterol. FIA determination of cholesterol with biosensors yielded a calibration curve with the following characteristics: linear range up to 50 μM, sensitivity of 43.5 nA μM?1 cm?2 and detection limit of 4.2 μM. The apparent Michaelis‐Menten constant and the response time of the biosensor are 0.49 mM and 15 s, respectively. This biosensor also exhibits good stability, reproducibility and long life time.  相似文献   

16.
《Electroanalysis》2018,30(2):274-282
Reduced Graphene oxide/ZnO nanoflowers ( rGO/ZnO‐NFs ) composite has been synthesized in‐situ using asymmetric Zn complex ( 1 ) as a single‐source molecular precursor (SSMP) with GO at 150 °C. The rGO/ZnO‐NFs composite was characterized by PXRD, UV‐vis, SEM, EDX mapping, TEM and SAED pattern to confirm its purity and morphology. The rGO/ZnO‐NFs composite shows uniform distribution of nanoflowers on graphene sheets. The modified glassy carbon electrode ( GCE ) was fabricated by drop wise layering of the rGO/ZnO‐NFs composite at the surface of the GCE without using binder. The binder free modified electrode ( GCE‐rGO/ZnO ) was explored for detection of nitroaromatics such as p‐nitro‐phenol ( p ‐NP ), 2,4‐dinitrophenol ( 2,4‐DNP ), 2,4‐dinitrotoluene ( 2,4‐DNT ) and 2,4,6‐trinitrophenol ( 2,4,6‐TNP ). The fabricated sensor showed remarkable response for the both toxicants and explosives. The LOD, sensitivity and linear range for the studied toxicants and explosives were found to be in a good range: p ‐NP= 0.93 μM, 240 μA mM−1 cm−2 and 0.2–0.9 mM; 2,4‐DNP= 6.2 μM, 203 μA mM−1 cm−2 and 0.1–0.9 mM; 2,4‐DNT= 10 μM, 371 μA mM−1 cm−2 and 0.2–0.9 mM; 2,4,6‐TNP= 16 μM, 514 μA mM−1 cm−2 and 0.2–0.9 mM, respectively.  相似文献   

17.
A choline biosensor was characterised in detail to determine the effects of physiologically relevant parameters on the ability of the sensor to reliably detect neurochemical changes in choline. This first generation Pt‐based polymer enzyme composite sensor displayed excellent shelf‐life and biocompatibility with no significant decrease in choline sensitivity observed following 14 days of storage dry, or in ex‐vivo rodent brain tissue. However, subjecting the sensor to repeated calibrations and storage over the same period resulted in significant decreases (20–70 %) due to enzyme denaturation associated with the repeated calibration and storage cycles. Potential interference signals generated by the principal electroactive interferents present in the brain were minimal; typically <1 % of the choline current response at in vivo levels. Additionally, changing temperature over the physiologically relevant range of 34–40 °C had no effect on sensitivity, while increasing pH between 7.2 and 7.6 produced only a 5 % increase in signal. The limit of detection of the sensor was in the low μM range (0.11±0.02 μM), while the in vitro response time was determined to be less than the solution mixing time and within ca. 5 s, suggesting potential sub‐second in vivo response characteristics. Finally, the sensor was implanted in the striatum of freely moving rats and demonstrated reliable detection of physiological changes in choline in response to movement, and pharmacological manipulation by injection of choline chloride.  相似文献   

18.
A chitosan (CS)‐tin oxide (SnO2) nanobiocomposite film has been deposited onto an indium‐tin‐oxide glass plate to immobilize cholesterol oxidase (ChOx) for cholesterol detection. The value of the Michaelis–Menten constant (Km) obtained as 3.8 mM for ChOx/CS‐SnO2/ITO is lower (8 mM) than that of a ChOx/CS/ITO bioelectrode revealing enhancement in affinity and/or activity of ChOx towards cholesterol and also revealing strong binding of ChOx onto CS‐SnO2/ITO electrode. This ChOx/CS‐SnO2/ITO cholesterol sensor retains 95% of enzyme activity after 4–6 weeks at 4 °C with response time of 5 s, sensitivity of 34.7 μA/mg dL?1 cm2 and detection limit of 5 mg/dL.  相似文献   

19.
A colloidal suspension of nanostructured poly(N-butyl benzimidazole)-graphene sheets (PBBIns-Gs) was used to modify a gold electrode to form a three-dimensional PBBIns-Gs/Au electrode that was sensitive to hydrogen peroxide (H2O2) in the presence of acetic acid (AcOH). The positively charged nanostructured poly(N-butyl benzimidazole) (PBBIns) separated the graphene sheets (Gs) and kept them suspended in an aqueous solution. Additionally, graphene sheets (Gs) formed “diaphragms” that intercalated Gs, which separated PBBIns to prevent tight packing and enhanced the surface area. The PBBIns-Gs/Au electrode exhibited superior sensitivity toward H2O2 relative to the PBBIns-modified Au (PBBIns/Au) electrode. Furthermore, a high yield of glucose oxidase (GOD) on the PBBIns-Gs of 52.3 mg GOD per 1 mg PBBIns-Gs was obtained from the electrostatic attraction between the positively charged PBBIns-Gs and negatively charged GOD. The non-destructive immobilization of GOD on the surface of the PBBIns-Gs (GOD-PBBIns-Gs) retained 91.5% and 39.2% of bioactivity, respectively, relative to free GOD for the colloidal suspension of the GOD-PBBIns-Gs and its modified Au (GOD-PBBIns-Gs/Au) electrode. Based on advantages including a negative working potential, high sensitivity toward H2O2, and non-destructive immobilization, the proposed glucose biosensor based on an GOD-PBBIns-Gs/Au electrode exhibited a fast response time (5.6 s), broad detection range (10 μM to 10 mM), high sensitivity (143.5 μA mM−1 cm−2) and selectivity, and excellent stability. Finally, a choline biosensor was developed by dipping a PBBIns-Gs/Au electrode into a choline oxidase (ChOx) solution for enzyme loading. The choline biosensor had a linear range of 0.1 μM to 0.83 mM, sensitivity of 494.9 μA mM−1 cm−2, and detection limit of 0.02 μM. The results of glucose and choline measurement indicate that the PBBIns-Gs/Au electrode provides a useful platform for the development of oxidase-based biosensors.  相似文献   

20.
Rate constants have been determined for the reactions of Cl atoms with the halogenated ethers CF3CH2OCHF2, CF3CHClOCHF2, and CF3CH2OCClF2 using a relative‐rate technique. Chlorine atoms were generated by continuous photolysis of Cl2 in a mixture containing the ether and CD4. Changes in the concentrations of these two species were measured via changes in their infrared absorption spectra observed with a Fourier transform infrared (FTIR) spectrometer. Relative‐rate constants were converted to absolute values using the previously measured rate constants for the reaction, Cl + CD4 → DCl + CD3. Experiments were carried out at 295, 323, and 363 K, yielding the following Arrhenius expressions for the rate constants within this range of temperature:Cl + CF3CH2OCHF2: k = (5.15 ± 0.7) × 10−12 exp(−1830 ± 410 K/T) cm3 molecule−1 s−1 Cl + CF3CHClOCHF2: k = (1.6 ± 0.2) × 10−11 exp(−2450 ± 250 K/T) cm3 molecule−1 s−1 Cl + CF3CH2OCClF2: k = (9.6 ± 0.4) × 10−12 exp(−2390 ± 190 K/T) cm3 molecule−1 s−1 The results are compared with those obtained previously for the reactions of Cl atoms with other halogenated methyl ethyl ethers. © 2001 John Wiley & Sons, Inc. Int J Chem Kinet 33: 165–172, 2001  相似文献   

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