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1.
《Electroanalysis》2006,18(5):456-464
An electrochemical DNA biosensor was developed by DNA immobilization at the electrode surface and its electrochemical behavior was studied in relation with different materials added in the paste. The aim was to study new materials for the development of new electrode surfaces, to be applied in the study of DNA – drug interactions. New electrochemical sensing materials using polymer multilayers were reported for the adsorption of DNA. These materials were prepared by mixing a polymer ion exchanger and graphite powder. The mixture was then used to render the modified carbon paste electrode (CPE), on the surface of which the dsDNA was adsorbed and studied by differential pulse voltammetry (DP voltammetry). The signal of guanine oxidation peak of DNA was followed. This modified biosensor was applied for the study of the interaction between DNA and the known intercalators Ethidium Bromide (EB) and Acridine Orange (AO). The established biosensor exhibited an improvement of its sensitivity and repeatability compared with the conventional CPE DNA biosensor.  相似文献   

2.
In this work, a novel multiplex electrochemiluminescence (ECL) DNA sensor has been developed for determination of hepatitis B virus (HBV) and hepatitis C virus (HCV) based on multicolor CdTe quantum dots (CdTe QDs) and Au nanoparticles (Au NPs). The electrochemically synthesized graphene nanosheets (GNs) were selected as conducting bridge to anchor CdTe QDs551-capture DNAHBV and CdTe QDs607-capture DNAHCV on the glassy carbon electrode (GCE). Then, different concentrations of target DNAHBV and target DNAHCV were introduced to hybrid with complementary CdTe QDs-capture DNA. Au NPs-probe DNAHBV and Au NPs-probe DNAHCV were modified to the above composite film via hybrid with the unreacted complementary CdTe QDs-capture DNA. Au NPs could quench the electrochemiluminescence (ECL) intensity of CdTe QDs due to the inner filter effect. Therefore, the determination of target DNAHBV and target DNAHCV could be achieved by monitoring the ECL DNA sensor based on Au NPs-probe DNA/target DNA/CdTe QDs-capture DNA/GNs/GCE composite film. Under the optimum conditions, the ECL intensity of CdTe QDs551 and CdTe QDs607 and the concentration of target DNAHBV and target DNAHCV have good linear relationship in the range of 0.0005–0.5 nmol L−1 and 0.001–1.0 nmol L−1 respectively, and the limit of detection were 0.082 pmol L−1 and 0.34 pmol L−1 respectively (S/N = 3). The DNA sensor showed good sensitivity, selectivity, reproducibility and acceptable stability. The proposed DNA sensor has been employed for the determination of target DNAHBV and target DNAHCV in human serum samples with satisfactory results.  相似文献   

3.
Du M  Yang T  Ma S  Zhao C  Jiao K 《Analytica chimica acta》2011,(2):9921-174
Electrochemical activities of typically electrochemical targets at three kinds of modified carbon electrodes, i.e. carbon ionic liquid electrode (CILE), graphene/carbon paste electrode (CPE), and ionic liquid-functionalized graphene (IL-graphene)/CPE, were compared in detail. The redox processes of the probes at IL-graphene/CPE were faster than those at CILE and graphene/CPE from cyclic voltammetry. An electrochemical method for the simultaneous determination of guanine and adenine was described with detection limits of 6.5 × 10−8 mol L−1 (guanine) and 3.2 × 10−8 mol L−1 (adenine). Single A → G mutation of sequence-specific DNA could be discriminated by the IL-graphene/CPE.  相似文献   

4.
We presented a new strategy for ultrasensitive detection of DNA sequences based on the novel detection probe which was labeled with Ag+ using metallothionein (MT) as a bridge. The assay relied on a sandwich-type DNA hybridization in which the DNA targets were first hybridized to the captured oligonucleotide probes immobilized on Fe3O4@Au composite magnetic nanoparticles (MNPs), and then the Ag+-modified detection probes were used to monitor the presence of the specific DNA targets. After being anchored on the hybrids, Ag+ was released down through acidic treatment and sensitively determined by a coupling flow injection–chemiluminescent reaction system (Ag+–Mn2+–K2S2O8–H3PO4–luminol) (FI–CL). The experiment results showed that the CL intensities increased linearly with the concentrations of DNA targets in the range from 10 to 500 pmol L−1 with a detection limit of 3.3 pmol L−1. The high sensitivity in this work may be ascribed to the high molar ratio of Ag+–MT, the sensitive determination of Ag+ by the coupling FI–CL reaction system and the perfect magnetic separation based on Fe3O4@Au composite MNPs. Moreover, the proposed strategy exhibited excellent selectivity against the mismatched DNA sequences and could be applied to real samples analysis.  相似文献   

5.
A simple and reliable method based on adsorptive stripping at an electrochemically pretreated glassy carbon electrode (GCE) was proposed for simultaneous or individual determination of guanine and adenine in DNA. The detection sensitivity of guanine and adenine was improved greatly by activating the GCE electrochemically. After accumulation on pretreated GCE at open circuit for 5 min or at the potential of +0.3 V for 120 s, guanine and adenine produced well-defined oxidation peaks at about +0.8 and +1.1 V, respectively in pH 5 phosphate buffer. The detection limit for individual measurement of guanine and adenine was 4.5 ng ml−1 (3×10−8 mol l−1) and 4 ng ml−1 (3×10−8 mol l−1), respectively. Acid-denatured DNA showed two oxidation peaks corresponding to guanine and adenine residues in the same buffer. The proposed method can be used to estimate the guanine and adenine contents in DNA with good selectivity in a linear range of 0.25-5 μg ml−1.  相似文献   

6.
A new electrochemical method has been proposed for the simultaneous determination of butylated hydroxyanisole (BHA) and propyl gallate (PG) in food matrices based on enzymatic biosensors. Spiny Au-Pt nanotubes (SAP NTs) was first synthesized and demonstrated to exhibit intrinsic peroxidase and catalase-like activity. The structure of SAP NTs provides large surface area and favorable medium for electron transfer, on which HRP were immobilized and acted as enzymatic biosensor for the simultaneous detection of BHA and PG. The results revealed that BHA and PG both have well-defined oxidation waves with peak potentials of 624 and 655 mV, respectively. Under the optimal conditions, the method behaved satisfactory analytical performance towards BHA and PG with a wide linear range of 0.3–50 mg L−1 and 0.1–100 mg L−1, as well as a detection limit of 0.046 mg L−1 and 0.024 mg L−1 (3σ/slope), respectively. Besides, the proposed method exhibits good sensitivity, stability and reproducibility, providing an alternative to fabricate electrode and construct sensitive biosensors.  相似文献   

7.
《Electroanalysis》2018,30(8):1659-1668
PAMAM dendrimer/reduced graphene oxide nanocomposite modified pencil graphite electrode (PAMAM/RGO/PGE) was used to fabricate an electrochemical DNA biosensor for determination of Rituxan (RTX) at low concentrations, for the first time. The fabricated biosensor was characterized with FE‐SEM, EIS, and CV techniques. The ds‐DNA/PAMAM/RGO/PGE was used as a working electrode to study the interaction between the RTX and salmon sperm ds‐DNA by DPV technique. Because of the interaction between the drug and DNA leads to a decrease in the guanine oxidation peak current, it was used as an indicator for the determination of the RTX. Under the optimized experimental conditions, a wide linear relationship between RTX concentration and guanine signal was obtained within the range of 7.0 to 60.0 μmol L−1 and 60.0 to 300.0 μmol L−1 with a low detection limit (0.56 μmol L−1). To clarify the interaction mechanism between the RTX and the ds‐DNA, DPV and UV‐Vis measurements were used. The reproducibility, stability, and performance of the constructed biosensor was examined by quantitative measuring RTX in pharmaceutical and human serum samples with good precision (RSD; 2.0–6.0 %) and acceptable recoveries (100.04–101.95 %).  相似文献   

8.
This work reports the advantages of a label free electrochemical aptasensor for the detection of lysozyme. The biorecognition platform was obtained by the adsorption of the aptamer on the surface of a carbon paste electrode (CPE) previously blocked with mouse immunoglobulin under controlled-potential conditions. The recognition event was detected from the decrease in the guanine and adenine electro-oxidation signals produced as a consequence of the molecular interaction between the aptamer and lysozyme. The biosensing platform demonstrated to be highly selective even in the presence of large excess (9-fold) of bovine serum albumin, cytochrome C and myoglobin. The reproducibility for 10 repetitive determinations of 10.0 mg L−1 lysozyme solution was 5.1% and 6.8% for guanine and adenine electro-oxidation signals, respectively. The detection limits of the aptasensor were 36.0 nmol L−1 (if considering guanine signal) and 18.0 nmol L−1 (if taking adenine oxidation current). This new sensing approach represents an interesting and promising alternative for the electrochemical quantification of lysozyme.  相似文献   

9.
A new strategy was introduced for ssDNA immobilization on a modified glassy carbon electrode. The electrode surface was modified using polyaniline and chemically reduced graphene oxide decorated cerium oxide nanoparticles (CeO2NPs-RGO). A single-stranded DNA (ssDNA) probe was immobilized on the modified electrode surface. Fast Fourier transform square wave voltammetry (FFT-SWV) was applied as detection technique and [Ru(bpy)3]2+/3+ redox signal was used as electrochemical marker. The hybridization of ssDNA with its complementary target caused a dramatic decrease in [Ru(bpy)3]2+/3+ FFT-SW signal. The proposed electrochemical biosensor was able to detect Aeromonas hydrophila DNA oligonucleotide sequence encoding aerolysin protein. Under optimal conditions, the biosensor showed excellent selectivity toward complementary sequence in comparison with noncomplementary and two-base mismatch sequences. The dynamic linear range of this electrochemical DNA biosensor for detecting 20-mer oligonucleotide sequence of A. hydrophila was from 1 × 10−15 to 1 × 10−8 mol L−1. The proposed biosensor was successfully applied for the detection of DNA extracted from A. hydrophila in fish pond water up to 0.01 μg mL−1 with RSD of 5%. Besides, molecular docking was applied to consider the [Ru(bpy)3]2+/3+ interaction with ssDNA before and after hybridization.  相似文献   

10.
The fast sequential multi-element determination of Ca, Mg, K, Cu, Fe, Mn and Zn in plant tissues by high-resolution continuum source flame atomic absorption spectrometry is proposed. For this, the main lines for Cu (324.754 nm), Fe (248.327 nm), Mn (279.482 nm) and Zn (213.857 nm) were selected, and the secondary lines for Ca (239.856 nm), Mg (202.582 nm) and K (404.414 nm) were evaluated. The side pixel registration approach was studied to reduce sensitivity and extend the linear working range for Mg by measuring at wings (202.576 nm; 202.577 nm; 202.578 nm; 202.580 nm; 202.585 nm; 202.586 nm; 202.587 nm; 202.588 nm) of the secondary line. The interference caused by NO bands on Zn at 213.857 nm was removed using the least-squares background correction. Using the main lines for Cu, Fe, Mn and Zn, secondary lines for Ca and K, and line wing at 202.588 nm for Mg, and 5 mL min− 1 sample flow-rate, calibration curves in the 0.1–0.5 mg L− 1 Cu, 0.5–4.0 mg L− 1 Fe, 0.5–4.0 mg L− 1 Mn, 0.2–1.0 mg L− 1 Zn, 10.0–100.0 mg L− 1 Ca, 5.0–40.0 mg L− 1 Mg and 50.0–250.0 mg L− 1 K ranges were consistently obtained. Accuracy and precision were evaluated after analysis of five plant standard reference materials. Results were in agreement at a 95% confidence level (paired t-test) with certified values. The proposed method was applied to digests of sugar-cane leaves and results were close to those obtained by line-source flame atomic absorption spectrometry. Recoveries of Ca, Mg, K, Cu, Fe, Mn and Zn in the 89–103%, 84–107%, 87–103%, 85–105%, 92–106%, 91–114%, 96–114% intervals, respectively, were obtained. The limits of detection were 0.6 mg L− 1 Ca, 0.4 mg L− 1 Mg, 0.4 mg L− 1 K, 7.7 µg L− 1 Cu, 7.7 µg L− 1 Fe, 1.5 µg L− 1 Mn and 5.9 µg L− 1 Zn.  相似文献   

11.
Natural matrices such as several vegetables, fruits or beverages are the origin of different natural antioxidants with great interest to be used instead of synthetic antioxidants in several applications, as a result of their health-promoting and disease-preventing properties. In this work, a capillary electrophoresis method capable of separating and quantifying some of these antioxidants (seven catechins with the same or similar molecular weight and relation q/z) is showed. Short analysis time and precision parameters equivalent to those obtained by liquid chromatography were obtained. This method was developed using the micellar electrokinetic chromatography (MEKC) mode with ultraviolet–visible detection. Quality parameters were established, obtaining low instrumental detection and quantification limits (0.6–2.0 mg L− 1 and 2.0–6.5 mg L− 1, respectively), good precision (relative standard deviation in the intermediate repeatability lower than 7% for every compound) and short analysis time. The developed method also showed good performance for the determination of two more natural antioxidants, quercetin and thymol. The applicability of the method to the analysis of catechins in several natural samples was tested.  相似文献   

12.
A syringe to syringe dispersive liquid phase microextraction-solidified floating organic drop was introduced and used for the simultaneous extraction of trace amounts of albendazole and triclabendazole from different matrices. The extracted analytes were determined by high performance liquid chromatography along with fluorescence detection. The analytical parameters affecting the microextraction efficiency including the nature and volume of the extraction solvent, sample volume, sample pH, ionic strength and the cycles of extraction were optimized. The calibration curves were linear in the range of 0.1–30.0 μg L−1 and 0.2–30.0 μg L−1 with determination coefficients of 0.9999 and 0.9998 for albendazole and triclabendazole respectively. The detection limits defined as three folds of the signal to noise ratio were found to be 0.02 μg L−1 for albendazole and 0.06 μg L−1 for triclabendazole. The inter-day and intra-day precision (RSD%) for both analytes at three concentration levels (0.5, 2.0 and 10.0 μg L−1) were in the range of 6.3–10.1% and 5.0–7.5% respectively. The developed method was successfully applied to determine albendazole and triclabendazole in water, cow milk, honey, and urine samples.  相似文献   

13.
A new diffusive gradients in a thin film (DGT) technique, using Microlite PrCH cation exchange resin, was developed and evaluated for measuring NH4–N in freshwaters. Microlite PrCH had high uptake (>92.5%) and elution efficiencies (87.2% using 2 mol L−1 NaCl). Mass vs. time validation experiments over 24 h demonstrated excellent linearity (R2 ≥ 0.996). PrCH-DGT binding layers had an extremely high intrinsic binding capacity for NH4–N (∼3000 μg). NH4–N uptake was quantitative over pH ranges 3.5–8.5 and ionic strength (up to 0.012 mol L−1 as NaCl) typical of freshwater systems. Several cations (Na+, K+, Ca2+ and Mg2+) were found to compete with NH4–N for uptake by PrCH-DGT, but NH4–N uptake was quantitative over concentration ranges typical of freshwater (up to 0.012 mol L−1 Na+, 0.006 mol L−1 K+, 0.003 mol L−1 Ca2+ and 0.004 mol L−1 Mg2+). Effective diffusion coefficients determined from mass vs. time experiments changed non-linearly with electrical conductivity. Field deployments of DGT samplers with varying diffusive layer thicknesses validated the use of the technique in situ, allowed deployment times to be manipulated with respect to NH4–N concentration, and enable the calculation of the diffusive boundary layer thickness. Daily grab sample NH4–N concentrations were observed to vary considerably independent of major rainfall events, but good agreements were obtained between PrCH-DGT values and mean grab sample measurements of NH4–N (CDGT:CSOLN 0.83–1.3). Reproducibility of DGT measurements in the field was good (relative standard deviation < 11%). Limit of detection was 0.63 μg L−1 (equivalent to 0.045 μmol L−1) based on 24 h deployments.  相似文献   

14.
In this paper, a novel colorimetric biosensor for Hg2+ and DNA molecules is presented based on Hg2+ stimulated oxidase-like activity of bovine serum albumin protected silver clusters (BSA-Ag NCs). Under mild conditions, Hg2+ activated BSA-Ag NCs to show high catalytic activity toward the oxidation of 3,3′,5, 5′-tetramethylbenzidine (TMB) using ambient dissolved oxygen as an oxidant. The oxidase-like activity of BSA-Ag NCs was “switched-on” selectively in the presence of Hg2+, which permitted a novel and facile colorimetric sensor for Hg2+. As low as 25 nmol L−1 Hg2+ could be detected with a linear range from 80 nmol L−1 to 50 mmol L−1. In addition, the sensing strategy was also employed to detect DNA molecules. Hg2+ is known to bind very strongly and specifically with two DNA thymine bases (T) to form thymine–Hg2+–thymine (T–Hg2+–T) base pairs. The hairpin-structure was disrupted and Hg2+ ions were released after hybridization with the DNA target. By coupling the Hg2+ switched-on the oxidase-mimicking activity of BSA-Ag NCs, we developed a novel label-free strategy for facile and fast colorimetric detection of DNA molecules. More important, target DNA can be detected as low as 10 nmol L−1 with a linear range from 30 to 225 nmol L−1. Compared with other methods, this method presents several advantages such as the independence of hydrogen peroxide, high sensitivity and good selectivity, avoiding any modification or immobilization of DNA, which holds a great potential of metal NCs for clinical application in biosensing and biotechnology.  相似文献   

15.
New voltammetric DNA biosensor based on a microcrystalline natural graphite–polystyrene composite film in the role of a transducer was used for the investigation of the interaction between model carcinogenic substance (2-aminofluorene; one of the most extensively studied examples of the aromatic amine class of carcinogens) and calf thymus double-stranded DNA (dsDNA). The layer of dsDNA immobilized at the electrode surface was utilized as a biocomponent responsive interface. The biosensor was characterized regarding the detection of DNA damage (induced by direct interaction with 2-aminofluorene) using square wave voltammetric responses of the guanine and adenine moieties and cyclic voltammetric responses of the anionic redox indicator [Fe(CN)6]4–/3– present in the solution.  相似文献   

16.
l-lactic acid is monitored during malolactic fermentation process of wine and its evolution is strongly related with the quality of the final product. The analysis of l-lactic acid is carried out off-line in a laboratory. Therefore, there is a clear demand for analytical tools that enabled real-time monitoring of this process in field and biosensors have positioned as a feasible alternative in this regard. The development of an amperometric biosensor for l-lactate determination showing long-term stability is reported in this work. The biosensor architecture includes a thin-film gold electrochemical transducer selectively modified with an enzymatic membrane, based on a three-dimensional matrix of polypyrrole (PPy) entrapping lactate oxidase (LOX) and horseradish peroxidase (HRP) enzymes. The experimental conditions of the biosensor fabrication regarding the pyrrole polymerization and the enzymes entrapment are optimized. The biosensor response to l-lactate is linear in a concentration range of 1 × 10−6–1 × 10−4 M, with a detection limit of 5.2 × 10−7 M and a sensitivity of – (13500 ± 600) μA M−1 cm−2. The biosensor shows an excellent working stability, retaining more than 90% of its original sensitivity after 40 days. This is the determining factor that allowed for the application of this biosensor to monitor the malolactic fermentation of three red wines, showing a good agreement with the standard colorimetric method.  相似文献   

17.
Quenchbody (Q-body) is a novel fluorescent biosensor based on the antigen-dependent removal of a quenching effect on a fluorophore attached to antibody domains. In order to develop a method using Q-body for the quantitative determination of deoxynivalenol (DON), a trichothecene mycotoxin produced by some Fusarium species, anti-DON Q-body was synthesized from the sequence information of a monoclonal antibody specific to DON. When the purified anti-DON Q-body was mixed with DON, a dose-dependent increase in the fluorescence intensity was observed and the detection range was between 0.0003 and 3 mg L−1. The coefficients of variation were 7.9% at 0.003 mg L−1, 5.0% at 0.03 mg L−1 and 13.7% at 0.3 mg L−1, respectively. The limit of detection was 0.006 mg L−1 for DON in wheat. The Q-body showed an antigen-dependent fluorescence enhancement even in the presence of wheat extracts. To validate the analytical method using Q-body, a spike-and-recovery experiment was performed using four spiked wheat samples. The recoveries were in the range of 94.9–100.2%. The concentrations of DON in twenty-one naturally contaminated wheat samples were quantitated by the Q-body method, LC-MS/MS and an immunochromatographic assay kit. The LC-MS/MS analysis showed that the levels of DON contamination in the samples were between 0.001 and 2.68 mg kg−1. The concentrations of DON quantitated by LC-MS/MS were more strongly correlated with those using the Q-body method (R2 = 0.9760) than the immunochromatographic assay kit (R2 = 0.8824). These data indicate that the Q-body system for the determination of DON in wheat samples was successfully developed and Q-body is expected to have a range of applications in the field of food safety.  相似文献   

18.
Saccharomyces cerevisiae immobilized in agarose gel as binding phase and polyacrylamide as diffusive layer in the diffusive gradient in thin films technique (DGT) was used for selective determination of methylmercury (MeHg). Deployment tests showed good linearity in mass uptake up to 48 h (3276 ng). When coupling the DGT technique with Cold Vapor Atomic Fluorescence Spectrometry, the method has a limit of detection of 0.44 ng L−1 (pre concentration factor of 11 for 48 h deployment). Diffusion coefficient of 7.03 ± 0.77 × 10−6 cm2 s−1 at 23 °C in polyacrylamide gel (pH = 5.5 and ionic strength = 0.05 mol L−1 NaCl) was obtained. Influence of ionic strength (from 0.0005 mol L−1 to 0.1 mol L−1 NaCl) and pH (from 3.5 to 8.5) on MeHg uptake were evaluated. For these range, recoveries of 84–105% and 84–98% were obtained for ionic strength and pH respectively. Potential interference due to presence of Cu, Fe, Mn, Zn was also assessed showing good recoveries (70–87%). The selectivity of the proposed approach was tested by deployments in solutions containing MeHg and Hg(II). Results obtained showed recoveries of 102–115 % for MeHg, while the uptake of Hg(II) was insignificant. The proposed approach was successfully employed for in situ measurements in the Negro River (Manaus-AM, Brazil).  相似文献   

19.
We describe a simple derivatization method to determine aldehydes. This method is based on derivatization with d-cysteine and consecutive liquid chromatography–tandem mass spectrometry (LC–MS/MS). The optimum derivatization conditions of aldehydes with d-cysteine were 10 min at 50 °C and pH 7.0. The formed alkyl thiazolidine-4-carboxylic acid derivatives were directly injected in LC–MS/MS. In the established condition, the method was used to detect eight aldehydes in beverages. The limit of detection (LOD) and limit of quantification (LOQ) of the aldehydes were 0.2–1.9 μg L−1 and 0.7–6.0 μg L−1 and the relative standard deviation was less than 2.0% at concentrations of 0.1 mg L−1 and 1.0 mg L−1 with the exception of octanal. All the beverage samples had detectable levels of methanal (0.033–0.145 mg L−1), ethanal (0.085–2.12 mg L−1), propanal (ND to 0.250 mg L−1), butanal (ND to 0.003 mg L−1), pentanal (ND to 0.471 mg L−1), hexanal (ND to 0.805 mg L−1), heptanal (0.019–3.91 mg L−1) and octanal (0.029–0.118 mg L−1).  相似文献   

20.
A novel approach is presented to determine mercury in urine samples, employing vortex-assisted ionic liquid dispersive liquid–liquid microextraction and microvolume back-extraction to prepare samples, and screen-printed electrodes modified with gold nanoparticles for voltammetric analysis. Mercury was extracted directly from non-digested urine samples in a water-immiscible ionic liquid, being back-extracted into an acidic aqueous solution. Subsequently, it was determined using gold nanoparticle-modified screen-printed electrodes. Under optimized microextraction conditions, standard addition calibration was applied to urine samples containing 5, 10 and 15 μg L−1 of mercury. Standard addition calibration curves using standards between 0 and 20 μg L−1 gave a high level of linearity with correlation coefficients ranging from 0.990 to 0.999 (N = 5). The limit of detection was empirical and statistically evaluated, obtaining values that ranged from 0.5 to 1.5 μg L−1, and from 1.1 to 1.3 μg L−1, respectively, which are significantly lower than the threshold level established by the World Health Organization for normal mercury content in urine (i.e., 10–20 μg L−1). A certified reference material (REC-8848/Level II) was analyzed to assess method accuracy finding 87% and 3 μg L−1 as the recovery (trueness) and standard deviation values, respectively. Finally, the method was used to analyze spiked urine samples, obtaining good agreement between spiked and found concentrations (recovery ranged from 97 to 100%).  相似文献   

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