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1.
A highly sensitive microchip electrophoresis (MCE) method with chemiluminescence (CL) detection was developed for the determination of biogenic amines including agmatine (Agm), epinephrine (E), dopamine (DA), tyramine, and histamine in human urine samples. To achieve a high assay sensitivity, the targeted analytes were pre-column labeled by a CL tagging reagent, N-(4-aminobutyl)-N-ethylisoluminol (ABEI). ABEI-tagged biogenic amines after MCE separation reacted with hydrogen peroxide in the presence of horseradish peroxidase (HRP), producing CL emission. Since no CL reagent was added to the running buffer, the background of the CL detection was extremely low, resulting in a significant improvement in detection sensitivity. Detection limits (S/N = 3) were in the range from 5.9 × 10−8 to 7.7 × 10−8 M for the biogenic amines tested, which were at least 10 times lower than those of the MCE–CL methods previously reported. Separation of a urine sample on a 7 cm glass/poly(dimethylsiloxane) (PDMS) microchip channel was completed within 3 min. Analysis of human urine samples found that the levels of Agm, E and DA were in the ranges of 2.61 × 10−7 to 4.30 × 10−7 M, 0.81 × 10−7 to 1.12 × 10−7 M, and 8.76 × 10−7 to 11.21 × 10−7 M (n = 4), respectively.  相似文献   

2.
A highly selective and sensitive electrogenerated chemiluminescence (ECL) biosensor for the detection of prostate PC-3 cancer cells was designed using a prostate specific antibody as a capture probe and ruthenium complex-labelled wheat germ agglutinin as a signal probe. The ECL biosensor was fabricated by covalently immobilising the capture probe on a graphene oxide-coated glassy carbon electrode. Target PC-3 cells were selectively captured on the surface of the biosensor, and then, the signal probe was bound with the captured PC-3 cells to form a sandwich. In the presence of tripropylamine, the ECL intensity of the sandwich biosensor was logarithmically directly proportion to the concentration of PC-3 cells over a range from 7.0 × 102 to 3.0 × 104 cells mL−1, with a detection limit of 2.6 × 102 cells mL−1. The ECL biosensor was also applied to detect prostate specific antigen with a detection limit of 0.1 ng mL−1. The high selectivity of the biosensor was demonstrated in comparison with that of a lectin-based biosensor. The strategy developed in this study may be a promising approach and could be extended to the design of ECL biosensors for highly sensitive and selective detection of other cancer-related cells or cancer biomarkers using different probes.  相似文献   

3.
A new electrogenerated chemiluminescence biosensor was fabricated by immobilizing ECL reagent Ru(bpy)32+ and alcohol dehydrogenase in sol-gel/chitosan/poly(sodium 4-styrene sulfonate) (PSS) organically modified composite material. The component PSS was used to immobilize ECL reagent Ru(bpy)32+ by ion-exchange, while the addition of chitosan was to prevent the cracking of conventional sol-gel-derived glasses and provide biocompatible microenvironment for alcohol dehydrogenase. Such biosensor combined enzymatic selectivity with the sensitivity of ECL detection for quantification of enzyme substrate and it was much simpler than previous double-layer design. The detection limit was 9.3 × 10−6 M for alcohol (S/N = 3) with a linear range from 2.79 × 10−5 to 5.78 × 10−2 M. With ECL detection, the biosensor exhibited wide linear range, high sensitivity and good stability.  相似文献   

4.
A reagentless signal-on electrochemiluminescence (ECL) biosensor for DNA hybridization detection was developed based on the quenching effect of ferrocene (Fc) on intrinsic cathodic ECL at thin oxide covered glassy carbon (C/CxO1−x) electrodes. To construct the DNA biosensor, molecular beacon (MB) modified with ferrocene (3′-Fc) was attached to a C/CxO1−x electrode via the covalent bound between labeled amino (5′-NH2) and surface functional groups. It was found that the immobilization of the probe on the electrode surface mainly depended on the fraction of surface carbonyl moiety. When a complementary target DNA (cDNA) was present, the stem-loop of MB on the electrode was converted into a linear double-helix configuration due to hybridization, resulting in the moving away of Fc from the electrode surface, and the restoring of the cathodic ECL signal. The restoration of the ECL intensity was linearly changed with the logarithm of cDNA concentration in the range of 1.0 × 10−11 to 7.0 × 10−8 M, and the detection limit was ca. 5.0 pM (S/N = 3). Additionally, single-base mismatched DNA can be effectively discriminated from the cDNA. The great advantage of the biosensor lies in its simplicity and cost-effective with ECL generated from the electrode itself, and no adscititious luminophore is required.  相似文献   

5.
Fakhari AR  Khorrami AR  Naeimi H 《Talanta》2005,66(4):813-817
A novel sensitive chromogenic reagent, N,N′-bis(3-methylsalicylidene)-ortho-phenylene diamine (MSOPD), has been synthesized and used in the spectrophotometric determination of nickel. At pH 8, MSOPD can react with nickel ion at room temperature to form a 1:1 complex. The apparent molar absorptivity is 9.5 × 104 l mol−1 cm−1 at 430 nm. Beer's low is obeyed over the range 0-1.0 × 10−5 M of nickel with a detection limit of 1.36 × 10−8 M. The relative standard deviation for measurement of 3.41 × 10−6 M nickel is 1.3% (n = 10). The method has successfully been applied to determination of trace amounts of nickel in some natural food samples.  相似文献   

6.
Zhang L  Li W  Shi M  Kong J 《Talanta》2006,70(2):432-436
A novel film modified electrode for the determination of trace lead was developed in this work. The modified electrode was prepared by the electropolymerization of N,N′-(o-phenylene)-bis-benzenesulfonamide (PBSA) as the ion capturing reagent to create the functional film. The modified electrode shows a high selectivity towards Pb2+ over interfering cations, e.g. Cu2+, Cd2+, Co2+, Ni2+, Zn2+, Cr2+, and the calibration curve was linear in the concentration range of 2.0 × 10−9 to 1.0 × 10−7 M with correlation coefficient of 0.999. For 20 min accumulation, detection limit of 1.0 × 10−9 M was obtained at the signal to noise ratio of 3. Analytical availability of the modified electrode was demonstrated by the application for samples from pond water.  相似文献   

7.
An electrochemiluminescence (ECL)-based method for rapid and sensitive detection of acridinium ester in neutral solution was described. Strong ECL emission was observed when a positive voltage over 2.0 V (versus Ag/AgCl) was applied to the working electrode (Pt) immersed in the acridinium ester solution of 2.0 mol l−1 KNO3 (pH 7.0). The possible ECL mechanism was discussed. It was proposed that the ECL emission came out of N-methylacridone, the oxidization product of acridinium ester by the nascent oxygen generated on the surface of working electrode in the course of oxidization of water. Other influenced factors including the electrochemical parameters, the ECL reaction medium and pH value, were investigated in detail. Under the optimal conditions, ECL intensity has a linear relationship with the concentration of acridinium ester in the range of 0.24-96 ng ml−1 (r=0.9999). The relative standard deviation for 24 ng ml−1 acridinium ester was 4.6% (n=11). The limit of detection was 0.16 ng ml−1.  相似文献   

8.
A new method for uric acid (UA) determination based on the quenching of the cathodic ECL of the tris(2,2-bipyridine)ruthenium(II)–uricase system is described. The biosensor is based on a double-layer design containing first tris(2,2-bipyridine)ruthenium(II) (Ru(bpy)32+) electrochemically immobilized on graphite screen-printed cells and uricase in chitosan as a second layer. The uric acid biosensing is based on the ECL quenching produced by uric acid over the cathodic ECL caused by immobilized Ru(bpy)32+ in the presence of uricase. The use of a −1.1 V pulse for 1 s with a dwelling time of 10 s makes it possible to estimate the initial enzymatic rate, which is used as the analytical signal. The Stern–Volmer type calibration function shows a dynamic range from 1.0 × 10−5 to 1.0 × 10−3 M with a limit of detection of 3.1 × 10−6 M and an accuracy of 13.6% (1.0 × 10−4 M, n = 5) as relative standard deviation. Satisfactory results were obtained for urine samples, creating an affordable alternative for uric acid determination.  相似文献   

9.
Two highly sensitive chemiluminescence (CL) systems are described. The method is based on the CL generated during the oxidation of luminol by N-bromosuccinimide (NBS) and N-chlorosuccinimide (NCS) in alkaline medium. The emission intensity is reduced by the presence of some surfactants at concentrations lower than critical micelle concentration (cmc).A new, simple, rapid and selective flow injection CL method for the determination of cationic surfactants such as dodecyltrimethylammonium bromide (DTAB), cetyltrimethylammonium bromide (CTAB) and cetylpyridinium chloride (CPC) is proposed. Their determinations are based on the reducing effect on the emission intensity of NBS-luminol and NCS-luminol chemiluminescent reactions. The effect of analytical and flow injection analysis (FIA) variables on these CL systems and on the determination of the cationic surfactants are discussed. The optimum parameters for the determination of cationic surfactants were studied and were found to be the following: luminol, 1×10−6 M; NBS and NCS both, 5×10−2 M; NaOH, 5×10−2 M and flow rate, 3.5 ml min−1.  相似文献   

10.
Solution studies on the binding properties of N-2,4-dimethylphenyl-N′-ethylformamidine (amitraz) toward nine lanthanide ions including lanthanum, cerium, neodium, samarium, europium, gadolinium, terbium, dysprosium, ytterbium and some other transition and heavy metal ions such as copper, lead, cobalt, nickel ions, showed a selective 1:1 complexation between amitraz and lanthanum ions. Consequently, amitraz was applied as an ion carrier in construction of a novel poly(vinyl chloride) membrane sensor for La(III). The sensor has a linear dynamic range of 1.0 × 10−1 to 1.0 × 10−7 M with a Nernstian slope of 19.8 ± 0.2 mV per decade and a detection limit of 8.0 × 10−8 M. The proposed sensor displays a fast response time (<8 s), and can be used for at least 2 months without any considerable divergences in the potentials. The La(III) membrane sensor revealed comparatively good selectivity with respect to most of cations including alkaline, alkaline earth, and some transition and heavy metal ions. It could be used in a pH range of 3.0-9.0. The proposed membrane electrode was used as an indicator electrode in the potentiometric titration of La(III) ions with an EDTA solution, and also in the determination of fluoride concentration in some mouth wash preparations.  相似文献   

11.
In this work, a novel La(III) membrane sensor based on 8-amino-N-(2-hydroxybenzylidene)naphthylamine (AIP) is presented. This electrode reveals good selectivity for La3+ over a wide variety of lanthanides metal ions. Theoretical calculations and conductance study of AIP to lanthanum and some other metal ions were carried out and confirmed selectivity toward La(III) ions. The electrode comprises 7% AIP, 30% PVC, 61% NPOE and 2% KTpClPB. The sensor displays a linear dynamic range between 1.0 × 10−7 and 1.0 × 10−1 M, with a nice Nernstian slope of 20.3 ± 0.3 mV per decade and a detection limit of 8.0 × 10−8 M. The potentiometric response is independent of pH in the range of 4.0-9.0. The proposed sensor posses the advantage of short response time, and especially, very good selectivity towards a large number of cations, such as Sm(III), Ce(III, Pr(III), Yb(III) and Hg(II), low detection limit and wide linear dynamic range in comparison with former ones. The electrode can be used for at least seven weeks without any considerable divergence in the potentials. It was used as an indicator electrode in the potentiometric titration of La(III) ions with EDTA. The sensor was applied to the determination of La(III) ions concentration in binary mixtures. It was also applied for the determination of fluoride ions in mouth wash preparations.  相似文献   

12.
Mehretie S  Admassie S  Hunde T  Tessema M  Solomon T 《Talanta》2011,85(3):1376-1382
A sensitive and selective method was developed for the determination of N-acetyl-p-aminophenol (APAP) and p-aminophenol (PAP) using poly(3,4-ethylenedioxythiophene) (PEDOT)-modified glassy carbon electrode (GCE). Cyclic voltammetry and differential pulse voltammetry were used to investigate the electrochemical reaction of APAP and PAP at the modified electrode. Both APAP and PAP showed quasireversible redox reactions with formal potentials of 367 mV and 101 mV (vs. Ag/AgCl), respectively, in phosphate buffer solution of pH 7.0. The significant peak potential difference (266 mV) between APAP and PAP enabled the simultaneous determination both species based on differential pulse voltammetry. The voltammetric responses gave linear ranges of 1.0 × 10−6-1.0 × 10−4 mol L−1 and 4.0 × 10−6-3.2 × 10−4 mol L−1, with detection limits of 4.0 × 10−7 mol L−1 and 1.2 × 10−6 mol L−1 for APAP and PAP, respectively. The method was successfully applied for the determination of APAP and PAP in pharmaceutical formulations and biological samples.  相似文献   

13.
A novel selective membrane electrode for determination of ultra-trace amount of lead was prepared. The PVC membrane containing N,N′-dimethylcyanodiaza-18-cown-6 (DMCDA18C6) directly coated on a graphite electrode, exhibits a Nernstian response for Pb2+ ions over a very wide concentration range (from 1.0×10−2 to 1.0×10−7 M) with a limit of detection of 7.0×10−8 M (∼14.5 ppb). It has a fast response time of ∼10 s and can be used for at least 2 months without any major deviation in potential. The electrode revealed very good selectivity with respect to all common alkali, alkaline earth, transition and heavy metal ions. The proposed sensor was used as an indicator electrode in potentiometric titration of lead ions and in determination of lead in edible oil, human hair and water samples. The proposed sensor was found to be superior to the best Pb2+-selective electrodes reported in terms of detection limit and selectivity coefficient.  相似文献   

14.
Qiu B  Xue L  Wu Y  Lin Z  Guo L  Chen G 《Talanta》2011,85(1):339-344
Inhibited Ru(bpy)32+ electrochemiluminescence by inorganic oxidants is investigated. Results showed that a number of inorganic oxidants can quench the ECL of Ru(bpy)32+/tri-n-propylamine (TPrA) system, and the logarithm of the decrease in ECL intensity (ΔI) was proportional to the logarithm of analyte concentrations. Based on which, a sensitive approach for detection of these inorganic oxidants was established, e.g. the log-log plots of ΔI versus the concentration of MnO4, Cr2O72− and Fe(CN)63− are linear in the range of 1 × 10−7 to 3 × 10−4 M for MnO4 and Cr2O72−, and 1 × 10−7 to 1 × 10−4 M for Fe(CN)63−, with the limit of detection (LOD) of 8.0 × 10−8 M, 2 × 10−8 M, and 1 × 10−8 M, respectively. A series of experiments such as a comparison of the inhibitory effect of different compounds on Ru(bpy)32+/TPrA ECL, ECL emission spectra, UV-Vis absorption spectra etc. were investigated in order to discover how these inorganic analytes quench the ECL of Ru(bpy)32+/TPrA system. A mechanism based on consumption of TPrA intermediate (TPrA·) by inorganic oxidants was proposed.  相似文献   

15.
Numerous drugs are carboxylic acid derivatives containing amino group, and hydrolysis reaction of these agents often generates toxic amines. Thus, the detection of amine impurity is of great importance in drug quality control of these amino group-containing ester and amide. A capillary electrophoresis method coupled with end-column electrochemiluminescent detection based on tris(2,2′-bipyridyl)ruthenium(II) system was proposed for the analysis of N,N-dimethyl ethanolamine (DMEA, the degradation product of meclophenoxate) in the presence of its precursor. Baseline separation of DMEA and meclophenoxate can be easily achieved under the selected conditions. DMEA can be assayed within 3 min over the concentration range of 5.0 × 10−8 to 3.0 × 10−6 mol L−1 with a detection limit of 2.0 × 10−8 mol L−1 at the signal-to-noise ratio of 3. The relative standard deviations of the signal intensity and the migration time were less than 5.3 and 2.5% for a standard sample containing 1.0 × 10−7 mol L−1 DMEA (n = 5), respectively. The presented method has been successfully applied for the profiling of DMEA resulting from the hydrolysis of meclophenoxate in commercial formulations. A primary stability investigation of meclophenoxate in aqueous solution was also carried out at different temperatures, and the results showed that the degradation of meclophenoxate accelerated at the higher temperature.  相似文献   

16.
Manuela Kim 《Talanta》2007,72(3):1054-1058
A simple and sensitive HPLC post-derivatization method with colorimetric detection has been developed for the determination of N-nitroso glyphosate in samples of technical glyphosate. Separation of the analyte was accomplished using an anionic exchange resin (2.50 mm × 4.00 mm i.d., 15 μm particle size, functional group: quaternary ammonium salt) with Na2SO4 0.0075 M (pH 11.5) (flow rate: 1.0 mL min−1) as mobile phase. After separation, the eluate was derivatized with a colorimetric reagent containing sulfanilamide 0.3% (w/v), [N-(1-naphtil)ethilendiamine] 0.03% (w/v) and HCl 4.5 M in a thermostatized bath at 95 °C. Detection was performed at 546 nm. All stages of the analytical procedure were optimized taking into account the concept of analytical minimalism: less operation times and costs; lower sample, reagents and energy consumption and minimal waste. The limit of detection (k = 3) calculated for 10 blank replicates was 0.04 mg L−1 (0.8 mg kg−1) in the solid sample which is lower than the maximum tolerable accepted by the Food and Agriculture Organization of the United Nations.  相似文献   

17.
Wang X  Zhou J  Yun W  Xiao S  Chang Z  He P  Fang Y 《Analytica chimica acta》2007,598(2):242-248
A sensitive and selective aptasensor using tri(2,2′-bipyridyl)ruthenium(II)-doped silica nanoparticles (Ru(bpy)32+-doped SNPs) as DNA tags for detection of thrombin is developed based on the target protein-induced strand displacement of the DNA probe. For the proposed aptasensor, the aptamer was assembled on the surface of the Au electrode through Au-S binding. The hybridization event between the DNA probe labeled by the Ru(bpy)32+-doped SNPs and the aptamer was evaluated by electrogenerated chemiluminescence (ECL) measurements. Then, the DNA probe was displaced by thrombin and the binding event between the thrombin and the aptamer was monitored by ECL measurements again. The difference of ECL intensity (ΔIECL) of the two events could be used to quantify the thrombin. Other proteins, such as bovine serum albumin and bovine hemoglobin, had almost negligible ΔIECL. Under the optimal conditions, the ΔIECL was linearly related to the concentration of the thrombin in the range of 10 fM to 10 pM and the detection limit was down to 1.0 fM since SNPs containing a large number of Ru(bpy)32+ molecules were labeled on the DNA probe.  相似文献   

18.
An electrochemiluminescent cholesterol disposable biosensor has been prepared by the formation of assembled layers on gold screen-printed cells. The detection layer is based on the electro-formation of new luminol copolymers with different synthesized biotinylated pyrroles prepared by click-chemistry, offering a new transduction layer with new electroluminescent properties on biosensors. The electrochemiluminescence (ECL) luminol copolymers are electroformed by cyclic voltammetry (five cycles) at pH 7.0 uses a10−3 M biotinylated pyrrole–luminol ratio of 1:10 in PBS buffer. With respect to the recognition layer, cholesterol oxidase was biotinylated by incubation with biotin vinyl sulfone, and immobilized on the copolymer by avidin–biotin interaction. The analytical signal of the biosensor is the ECL enzymatic initial rate working in chronoamperometric mode at 0.5 V excitation potential with 10 s between pulses at pH 9.5. The disposable device offers a cholesterol linear range from 1.5 × 10−5 M to 8.0 × 10−4 M with a limit of detection of 1.47 × 10−5 M and accuracy of 7.9% for 9.0 × 10−5 M and 14.1% for 2.0 × 10−4 M, (n = 5). Satisfactory results were obtained for cholesterol determination in serum samples compared to a reference procedure.  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.
Xiluan Yan  Masaaki Kai 《Talanta》2009,79(2):383-4519
We have developed a novel sensitive chemiluminescence (CL) aptasensor for the target assay as exemplified by using adenosine as a model target. In this work, we have demonstrated the signaling mechanism to make detection based on magnetic separation and 3,4,5-trimethoxyl-phenylglyoxal (TMPG), a special CL reagent as the signaling molecule, which reacts instantaneously with guanine nucleobases (G) of adenosine-binding aptamer strands. Briefly, amino-functioned capture DNA sequences are immobilized on the surface of carboxyl-modified magnetic beads, and then hybridized with label-free G-rich (including 15 guanine nucleobases) adenosine-binding aptamer strands to form our CL aptasensor. Upon the introduction of adenosine, the aptamer on the surface of magnetic beads is triggered to make structure switching to the formation of the adenosine/aptamer complex. Consequently, G-rich aptamer strands are forced to dissociate from magnetic beads sensing interface, resulting in a decrease of CL signal. The decrement of peak signal is proportional to the amount of adenosine. The effects of the amounts of capture DNA, aptamer, magnetic beads are investigated and optimized. It was found that the CL intensity had a linear dependency on the concentration of adenosine in the range of 4 × 10−7 to 1 × 10−5 M. With a low detection limit of 8 × 10−8 M and simplicity in CL detection, this novel technique will offer a great promise for future target/aptamer analysis.  相似文献   

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