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1.
A novel amperometric biosensor utilizing two enzymes, glucose oxidase (GOD) and horseradish peroxidase (HRP), was developed for the cathodic detection of glucose. The glucose biosensor was constructed by electrochemical formation of a polypyrrole (PPy) membrane in the presence of GOD on the surface of a HRP-modified sol-gel derived-mediated ceramic carbon electrode. Ferrocenecarboxylic acid (FCA) was used as mediator to transfer electron between enzyme and electrode. In the hetero-bilayer configuration of electrode, all enzymes were well immobilized in electrode matrices and showed favorable enzymatic activities. The amperometric detection of glucose was carried out at +0.16 V (versus saturated calomel reference electrode (SCE)) in 0.1 M phosphate buffer solution (pH 6.9) with a linear response range between 8.0×10−5 and 1.3×10−3 M glucose. The biosensor showed a good suppression of interference in the amperometric detection.  相似文献   

2.
Qu F  Shi A  Yang M  Jiang J  Shen G  Yu R 《Analytica chimica acta》2007,605(1):28-33
Prussian blue nanowire array (PBNWA) was prepared via electrochemical deposition with polycarbonate membrane template for effective modification of glassy carbon electrode. The PBNWA electrode thus obtained was demonstrated to have high-catalytic activity for the electrochemical reduction of hydrogen peroxide in neutral media. This enabled the PBNWA electrode to show rapid response to H2O2 at a low potential of −0.1 V over a wide range of concentrations from 1 × 10−7 M to 5 × 10−2 M with a high sensitivity of 183 μA mM−1 cm−2. Such a low-working potential also substantially improved the selectivity of the PBNWA electrode against most electroactive species such as ascorbic acid and uric acid in physiological media. A detection limit of 5 × 10−8 M was obtained using the PBNWA electrode for H2O2, which compared favorably with most electroanalysis procedures for H2O2. A biosensor toward glucose was then constructed with the PBNWA electrode as the basic electrode by crosslinking glucose oxidase (GOx). The glucose biosensor allowed rapid, selective and sensitive determination of glucose at −0.1 V. The amperometric response exhibited a linear correlation to glucose concentration through an expanded range from 2 × 10−6 M to 1 × 10−2 M, and the response time and detection limit were determined to be 3 s and 1 μM, respectively.  相似文献   

3.
Multiplex electrochemical detection of two DNA target sequences in one sample using enzyme-functionalized Au nanoparticles (AuNPs) as catalytic labels for was proposed. This DNA sensor was fabricated using a “sandwich” detection strategy, involving two kinds of capture probes DNA immobilized on glassy carbon electrode (GCE), and hybridization with target DNA sequences, which further hybridized with the reporter DNA loaded on the AuNPs. The AuNP contained two kinds of DNA sequences, one was complementary to the target DNA, while the other was noncomplementary to the target. The noncomplementary sequences were linked with horseradish peroxidase (HRP) and alkaline phosphatase (ALP), respectively. Enhanced detection sensitivity was obtained where the AuNPs carriers increased the amount of enzyme molecules per hybridization. Electrochemical signals were generated from the enzymatic products produced from the substrates catalyzed by HRP and ALP. Under optimal conditions, a 33-mer sequence could be quantified over the ranges from 1.5 × 10−13 to 5.0 × 10−12 M with a detection limit of 1.0 × 10−13 M using HRP-AuNP as labels, and a 33-mer sequence could be quantified over the ranges from 4.5 × 10−11 M to 1.0 × 10−9 M with a detection limit of 1.2 × 10−11 M using ALP-AuNP as labels.  相似文献   

4.
The harmful effects of cyanotoxins in surface waters have led to increasing demands for accurate early warning methods. This study proposes a capacitive immunosensor for broad-spectrum detection of the group of toxic cyclic peptides called microcystins (∼80 congeners) at very low concentration levels. The novel analytical platform offers significant advances compared to the existing methods. Monoclonal antibodies (mAbs, clone AD4G2) that recognize a common element of microcystins were used to construct the biosensing layer. Initially, a stable insulating anchor layer for the mAbs was made by electropolymerization of tyramine onto a gold electrode surface, with subsequent incorporation of gold nanoparticles (AuNPs) on the glutaraldehyde (5%) activated polytyramine surface. The biosensor responded linearly to microcystin concentrations from 1 × 10−13 M to 1 × 10−10 M MC-LR standard with a limit of detection of 2.1 × 10−14 M. The stability of the biosensor was evaluated by repeated measurements of the antigen and by determining the capacitance change relative to the original response, which decreased below 90% after the 30th cycle.  相似文献   

5.
Jianwen Wang  Yifeng Tu 《Talanta》2009,77(4):1454-4466
A novel disposable third-generation hydrogen peroxide (H2O2) biosensor based on horseradish peroxidase (HRP) immobilized on the gold nanoparticles (AuNPs) electrodeposited indium tin oxide (ITO) electrode is investigated. The AuNPs deposited on ITO electrode were characterized by UV-vis, SEM, and electrochemical methods. The AuNPs attached on the ITO electrode surface with quasi-spherical shape and the average size of diameters was about 25 nm with a quite symmetric distribution. The direct electron chemistry of HRP was realized, and the biosensor exhibited excellent performances for the reduction of H2O2. The amperometric response to H2O2 shows a linear relation in the range from 8.0 μmol L−1 to 3.0 mmol L−1 and a detection limit of 2 μmol L−1 (S/N = 3). The value of HRP immobilized on the electrode surface was found to be 0.4 mmol L−1. The biosensor indicates excellent reproducibility, high selectivity and long-term stability.  相似文献   

6.
We first reported an ultrasensitive hydrogen peroxide biosensor in this work. The biosensor was fabricated by coating graphene–gold nanocomposite (G–AuNP), CdTe–CdS core–shell quantum dots (CdTe–CdS), gold nanoparticles (AuNPs) and horseradish peroxidase (HRP) in sequence on the surface of gold electrode (GE). Cyclic voltammetry and differential pulse voltammetry were used to investigate electrochemical performances of the biosensor. Since promising electrocatalytic synergy of G–AuNP, CdTe–CdS and AuNPs towards hydrogen peroxide was achieved, the biosensor displayed a high sensitivity, low detection limit (S/N = 3) (3.2 × 10−11 M), wide calibration range (from 1 × 10−10 M to 1.2 × 10−8 M) and good long-term stability (20 weeks). Moreover, the effects of omitting G–AuNP, CdTe–CdS and AuNP were also examined. It was found that sensitivity of the biosensor is more 11-fold better if G–AuNP, CdTe–CdS and AuNPs are used. This could be ascribed to improvement of the conductivity between graphene nanosheets in the G–AuNP due to introduction of the AuNPs, ultrafast charge transfer from CdTe–CdS to the graphene sheets and AuNP due to unique electrochemical properties of the CdTe–CdS, and good biocompatibility of the AuNPs for horseradish peroxidase. The biosensor is of best sensitivity in all hydrogen peroxide biosensors based on graphene and its composites up to now.  相似文献   

7.
A sensitive non-enzymatic glucose electrochemical biosensor (Cu/PMo12-GR/GCE) was developed based on the combination of copper nanoparticles (CuNPs) and phosphomolybdic acid functionalized graphene (PMo12-GR). PMo12-GR films were modified on the surface of glassy carbon electrode (GCE) through electrostatic self-assembly with the aid of poly diallyl dimethyl ammonium chloride (PDDA). Then CuNPs were successfully decorated onto the PMo12-GR modified GCE through electrodeposition. The morphology of Cu/PMo12-GR/GCE was characterized by scanning electron microscope (SEM). Cyclic voltammetry (CV) and chronoamperometry were used to investigate the electrochemical performances of the biosensor. The results indicated that the modified electrode displayed a synergistic effect of PMo12-GR sheets and CuNPs towards the electro-oxidation of glucose in the alkaline solution. At the optimal detection potential of 0.50 V, the response towards glucose presented a linear response ranging from 0.10 μM to 1.0 mM with a detection limit of 3.0 × 10−2 μM (S/N = 3). In addition, Cu/PMo12-GR/GCE possessed a high selectivity, good reproducibility, excellent stability and acceptable recovery, which indicating the potential application in clinical field.  相似文献   

8.
An electrochemiluminescence (ECL) biosensor for simultaneous detection of adenosine and thrombin in one sample based on bifunctional aptamer and N-(aminobutyl)-N-(ethylisoluminol) functionalized gold nanoparticles (ABEI-AuNPs) was developed. A streptavidin coated gold nanoparticles modified electrode was utilized to immobilize biotinylated bifunctional aptamer (ATA), which consisted of adenosine and thrombin aptamer. The ATA performed as recognition element of capture probe. For adenosine detection, ABEI-AuNPs labeled hybridization probe with a partial complementary sequence of ATA reacted with ATA, leading to a strong ECL response of N-(aminobutyl)-N-(ethylisoluminol) enriched on ABEI-AuNPs. After recognition of adenosine, the hybridization probe was displaced by adenosine and ECL signal declined. The decrease of ECL signal was in proportion to the concentration of adenosine over the range of 5.0 × 10−12–5.0 × 10−9 M with a detection limit of 2.2 × 10−12 M. For thrombin detection, thrombin was assembled on ATA modified electrode via aptamer–target recognition, another aptamer of thrombin tagged with ABEI-AuNPs was bounded to another reactive site of thrombin, producing ECL signals. The ECL intensity was linearly with the concentration of thrombin from 5 × 10−14 M to 5 × 10−10 M with a detection limit of 1.2 × 10−14 M. In the ECL biosensor, adenosine and thrombin can be detected when they coexisted in one sample and a multi-analytes assay was established. The sensitivity of the present biosensor is superior to most available aptasensors for adenosine and thrombin. The biosensor also showed good selectivity towards the targets. Being challenged in real plasma sample, the biosensor was confirmed to be a good prospect for multi-analytes assay of small molecules and proteins in biological samples.  相似文献   

9.
Fengna Xi 《Talanta》2009,78(3):1077-360
A simple and controllable electrodeposition approach was established for one-step construction of novel reagentless biosensors by in situ formation of chitosan-carbon nanotubes-nile blue-horseradish peroxidase (CS-CNTs-NB-HRP) biocomposite film on electrode surface. The mediator effect of NB, conducting performance of CNTs and the biocompatible microenvironment of CS were combined by such one-step non-manual process. NB could interact with CNTs and resulted in good dispersion of CNTs-NB nanocomposites in aqueous solution. Cyclic voltammetry measurements demonstrated that electrons were efficiently shuttled between HRP and the electrode mediated by NB. The developed reagentless biosensor exhibited a fast amperometric response for the determination of H2O2 and 95% of the steady-state current was obtained within 2 s. The linear response of the reagentless biosensor for the determination of H2O2 ranged from 1.0 × 10−6 to 2.4 × 10−4 mol l−1 with a detection limit of 1.2 × 10−7 mol l−1. The biosensor exhibited high reproducibility and long-time storage stability. The as-prepared biosensor also showed effective anti-interference capability. The ease of the one-step non-manual technique and the promising feature of the biocomposite could serve as a versatile platform for fabricating electrochemical biosensors.  相似文献   

10.
We here reported a simple electrochemical method for the detection of tryptophan (Trp) based on the Ag@C modified glassy carbon (Ag@C/GC) electrode. The Ag@C core–shell structured nanoparticles were synthesized using one-pot hydrothermal method and characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), and Fourier transform-infrared spectroscopy (FTIR). The electrochemical behaviors of Trp on Ag@C/GC electrode were investigated and exhibited a direct electrochemical process. The favorable electrochemical properties of Ag@C/GC electrode were attributed to the synergistic effect of the Ag core and carbon shell. The carbon shell cannot only protect Ag core but also contribute to the enhanced substrate accessibility and Trp-substrate interactions, while nano-Ag core can display good electrocatalytic activity to Trp at the same time. Under the optimum experimental conditions the oxidation peak current was linearly dependent on the Trp concentration in the range of 1.0 × 10−7 to 1.0 × 10−4 M with a detection limit of 4.0 × 10−8 M (S/N = 3). In addition, the proposed electrode was applied for the determination of Trp concentration in real samples and satisfactory results were obtained. The technique offers enhanced sensitivity and may trigger the possibilities of the Ag@C nanocomposite towards diverse applications in biosensor and electroanalysis.  相似文献   

11.
A convenient microwave plasma treatment method with ammonia precursor was proposed to enhance the solubility of carbon nanotubes (CNTs). The SEM, XRD and FTIR spectra clearly demonstrated that the carbon skeleton structure of the resultant ammonia plasma-treated CNTs (ammonia PT-CNTs) was not destroyed and amine groups of different forms were successfully coupled to CNTs in the MWP treatment process. The ammonia PT-CNTs have excellent solubility in water and are insoluble in nonpolar tetrahydrofuran, and the cyclic voltammograms suggest that the enhanced wetting properties clearly favor faster electron transfer kinetics on the ammonia PT-CNT electrodes. By choosing glucose oxidase as a model enzyme, the application of the ammonia PT-CNTs in construction of biosensors was further investigated. Due to the biocompatibility and electron transfer capability of the ammonia PT-CNTs, the resultant GOD biosensor displayed a good sensing performance. The biosensor has a fast response of less than 10 s, and the response current linearly increases with the glucose concentration in the range of 1.2 × 10−4 to 7.5 × 10−3 M with a detection limit of 1.0 × 10−5 M.  相似文献   

12.
The remarkable synergistic effects of the zinc oxide (ZnO) nanoparticles and multi-walled carbon nanotubes (MWNTs) were developed for the ssDNA probe immobilization and fabrication of the electrochemical DNA biosensor. The ZnO/MWNTs/chitosan nanocomposite membrane-modified glassy carbon electrode (ZnO/MWNTs/CHIT/GCE) was fabricated and the ssDNA probes were immobilized on the modified electrode surface. The preparation method is quite simple and inexpensive. The hybridization events were monitored by differential pulse voltammetry (DPV) using methylene blue (MB) as an indicator. As compared with previous MWNTs-based DNA biosensors, this composite matrix combined the attractive biocompatibility of ZnO nanoparticles with the excellent electron-transfer ability of MWNTs and fine membrane-forming ability of CHIT increased the DNA attachment quantity and complementary DNA detection sensitivity. The approach described here can effectively discriminate complementary DNA sequence, noncomplementary sequence, single-base mismatched sequence and double-base mismatched sequence related to phosphinothricin acetyltransferase (PAT) gene in transgenic corn. Under optimal conditions, the dynamic detection range of the sensor to PAT gene complementary target sequence was from 1.0 × 10−11 to 1.0 × 10−6 mol/L with the detection limit of 2.8 × 10−12 mol/L. The polymerase chain reaction (PCR) amplification of nopaline synthase (NOS) gene from the real sample of one kind of transgenic soybeans was also satisfactorily detected with this electrochemical DNA biosensor, suggesting that the ZnO/MWNTs/CHIT nanocomposite hold great promises for sensitive electrochemical biosensor applications.  相似文献   

13.
In this work, we reported an efficient platinum nanoparticles functionalized nitrogen doped graphene (PtNPs@NG) nanocomposite for devising novel electrochemical glucose biosensor for the first time. The fabricated PtNPs@NG and biosensor were characterized using transmission electron microscopy, high-resolution transmission electron microscopy, X-ray photoelectron spectroscopy, static water contact angle, UV–vis spectroscopy, electrochemical impedance spectra and cyclic voltammetry, respectively. PtNPs@NG showed large surface area and excellent biocompatibility, and enhanced the direct electron transfer between enzyme molecules and electrode surface. The glucose oxidase (GOx) immobilized on PtNPs@NG nanocomposite retained its bioactivity, and exhibited a surface controlled, quasi-reversible and fast electron transfer process. The constructed glucose biosensor showed wide linear range from 0.005 to 1.1 mM with high sensitivity of 20.31 mA M−1 cm−2. The detection limit was calculated to be 0.002 mM at signal-to-noise of 3, which showed 20-fold decrease in comparison with single NG-based electrochemical biosensor for glucose. The proposed glucose biosensor also demonstrated excellent selectivity, good reproducibility, acceptable stability, and could be successfully applied in the detection of glucose in serum samples at the applied potential of −0.33 V. This research provided a promising biosensing platform for the development of excellent electrochemical biosensors.  相似文献   

14.
This paper proposed a novel method for ultra-trace detection of pesticides combining electrochemical reduction of Ellman's reagent with acetylcholinesterase (AChE) inhibition. The amperometric biosensor, fabricated by immobilizing AChE on multi-walled carbon nanotubes-chitosan (MWCNTs-Chi) nanocomposites modified glassy carbon electrode, enjoyed high sensitivity owing to the excellent conductivity and favourable biocompatibility of MWCNTs-Chi nanocomposites. Meanwhile, the sensitivity of the biosensor was further enhanced using the electrochemical reduction signal of DTNB for determination. Under optimum conditions, methyl parathion was detected based on its inhibition effect on AChE activity and the subsequent change in electrochemical reduction response of DTNB. Good relationship was obtained between the reduction current and pesticide concentration in the ranges of 5.0 × 10−7 to 1.0 × 10−12 M with a detection limit of 7.5 × 10−13 M (S/N = 3). Moreover, the proposed protocol was successfully employed for the determination of methyl parathion in water and soil samples.  相似文献   

15.
A new palladium nanoparticle functionalized multi-wall carbon nanotubes (nano-Pd/CNTs) modified pyrolytic graphite electrode (PGE) has been fabricated for electrochemical sensing of calcium dobesilate (CD) in pharmaceutical capsules. The nano-Pd/CNTs were characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). The nano-Pd/CNTs composite showed a strong electrocatalytic property for CD. The anodic peak current is 6-fold than that obtained in bare PGE and the oxidation potential has an obvious shift to negative. The anodic peak current is proportional to the concentration of CD in the range of 1.0 × 10−7 to 7.0 × 10−4 mol L−1, with a linear relative coefficient r = 0.999 and a detection limit 4.0 × 10−8 mol L−1 (S/N = 3). This kind of electrode shows good stability, sensitivity, reproducibility, large linear range and low detection limit towards electrochemical determination of CD. The proposed method provides a selective and sensitive electrochemical sensor of calcium dobesilate.  相似文献   

16.
Carbon nanoparticles (CNPs) and halloysite nanoclay (HNC) modified carbon paste electrode (HNC–CNP–CPE) was developed for the determination of methyl parathion (MP) and ethyl parathion (EP). The electrochemical behavior of these molecules was investigated employing cyclic voltammetry (CV), chronocoulometry (CC), electrochemical impedance spectroscopy (EIS) and potentiometric stripping analysis (PSA). After optimization of analytical conditions employing this electrode at pH 5.0 in acetate buffer (0.1 M), the peak currents were found to vary linearly with its concentration in the range of 1.55 × 10−9 to 3.67 × 10−6 M and 1.21 × 10−9 to 4.92 × 10−6 M for MP and EP, respectively. The detection limits (S/N = 3) of 4.70 × 10−10 M and 3.67 × 10−10 M were obtained for MP and EP, respectively, using PSA. The prepared modified electrode showed several advantages such as simple preparation method, high sensitivity, very low detection limits and excellent reproducibility. The proposed method was employed for the determination of MP and EP in fruits, vegetables, water and soil samples.  相似文献   

17.
Miaomiao Gu 《Talanta》2009,80(1):246-1985
Gold nanoparticles (AuNPs) were assembled on the surface of polystyrene (PS) and polyaniline (PANI) core-shell nanocomposite (PS@PANI) for the immobilization of HL-60 leukemia cells to fabricate a cell electrochemical sensor. The immobilized cells exhibited irreversible voltammetric response and increased the electron transfer resistance with a good correlation to the logarithmic value of concentration ranging from 1.6 × 103 to 1.6 × 108 cells mL−1 with a limit of detection of 7.3 × 102 cells mL−1 at 10σ. This biosensor was simple, low cost and disposable, which implied that the PS@PANI/Au composites can regard as the potential applications for clinical applications.  相似文献   

18.
Chen S  Du D  Huang J  Zhang A  Tu H  Zhang A 《Talanta》2011,84(2):451-456
An electrochemical strategy on the basis of rationally designed molecularly imprinted sol-gel polymer embedded with gold nanoparticles (AuNPs) is developed for the specific and sensitive determination of Sudan I. The rationally designed sensing Sudan I imprinted sol-gel was prepared by mixing Sudan I with 3-aminopropyltriethoxysilane, tetraethoxysilane, chitosan, and AuNPs, followed by copolymerization and extraction of the template molecules. The hybrid forming membrane was characterized by SEM and FTIR-ATR, and used for the linear sweep voltammetric (LSV) determination of Sudan I in water/ethanol solutions. The LSV responses exhibited high sensitivity and selectivity, as discriminated from Sudan I analogues. Under optimal experimental conditions, LSV peak currents were linearly proportional to the concentrations of Sudan I in the range from 0.1 × 10−7 to 1.0 × 10−5 M, with a detection limit of 2.0 × 10−9 M. The strategy is generally applicable in developing sensitive, selective, and moreover, reusable electrochemical sensors for quantitative determination of electroactive species.  相似文献   

19.
A novel and sensitive electrochemical DNA biosensor has been developed for the detection of DNA hybridization. The biosensor was proposed by using copper(II) complex of Luteolin C30H18CuO12 (CuL2) as an electroactive indicator based on silver nanoparticles and multi-walled carbon nanotubes (Ag/MWCNTs) modified glassy carbon electrode (GCE). In this method, the 4-aminobenzoic acid (4-ABA) and Ag nanoparticles were covalently grafted on MWCNTs to form Ag/4-ABA/MWCNTs. The proposed method dramatically increased DNA attachment quantity and complementary ssDNA detection sensitivity for its large surface area and good charge-transport characteristics. DNA hybridization detection was performed using CuL2 as an electroactive indicator. The CuL2 was synthesized and characterized using elemental analysis (EA) and IR spectroscopy. Cyclic voltammetry (CV) and fluorescence spectroscopy were used to investigate the interaction between CuL2 and ds-oligonucleotides (dsDNA). It was revealed that CuL2 presented high electrochemical activity on GCE, and it could be intercalated into the double helices of dsDNA. The target ssDNA of the human hepatitis B virus (HBV) was quantified in a linear range from 3.23 × 10−12 to 5.31 × 10−9 M (r = 0.9983). A detection limit of 6.46 × 10−13 M (3σ, n = 11) was achieved.  相似文献   

20.
For the first time, a new platform based on electrochemical growth of Au nanoparticles on aligned multi-walled carbon nanotubes (A-MWCNT) was developed for sensitive lable-free DNA detection of the TP53 gene mutation, one of the most popular genes in cancer research. Electrochemical impedance spectroscopy (EIS) was used to monitor the sequence-specific DNA hybridization events related to TP53 gene. Compared to the bare Ta or MWCNT/Ta electrodes, the synergistic interactions of vertically aligned MWCNT array and gold nanoparticles at modified electrode could improve the density of the probe DNA attachment and resulting the sensitivity of the DNA sensor greatly. Using EIS, over the extended DNA concentration range, the change of charge transfer resistance was found to have a linear relationship in respect to the logarithm of the complementary oligonucleotides sequence concentrations in the wide range of 1.0 × 10−15 − 1.0 × 10−7 M, with a detection limit of 1.0 × 10−17 M (S/N = 3). The prepared sensor also showed good stability (14 days), reproducibility (RSD = 2.1%) and could be conveniently regenerated via dehybridization in hot water. The significant improvement in sensitivity illustrates that combining gold nanoparticles with the on-site fabricated aligned MWCNT array represents a promising platform for achieving sensitive biosensor for fast mutation screening related to most human cancer types.  相似文献   

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