Highly sensitive deoxynivalenol immunosensor based on a glassy carbon electrode modified with a fullerene/ferrocene/ionic liquid composite

We describe a sensitive electrochemical immunosensor for the detection of deoxynivalenol (DON). It is based on a glassy carbon electrode modified with a composite made from fullerene (C₆₀), ferrocene and the ionic liquid. The components were immobilized on the surface of the electrode using chitosan cross-linked with epichlorohydrin. Then, the antibody to DON was covalently conjugated to the surface which then was blocked with serum albumin. The performance of the immunosensor was investigated by cyclic voltammetry and electrochemical impedance spectroscopy. It offers good repeatability (RSD?=?1.2%), selectivity, a stability of more than 180?days, an impedimetric response to DON in the range of 1?pgmL^?1 to 0.3?ng?mL^?1, and a detection limit (at S/N?=?3) of 0.3?pgmL^?1. The limit of detection is better than that of GC, HPLC, GC-MS, HPLC-MS and LC-MS-MS. The effects of omitting C₆₀ or the ionic liquid were also examined. The results indicate that the sensitivity of the biosensor is 2-fold better if C₆₀ and ionic liquids are used. This demonstrates that C₆₀ facilitates electron transfer on the surface of the modified electrode due to its unique electrochemical properties, while the ionic liquid provides a biocompatible microenvironment for the antibody. This results in increased sensitivity and stability. The method was satisfactorily applied to the determination of DON in food samples.

Cetyltrimethylammonium-coated magnetic nanoparticles for the extraction of bromate,followed by its spectrophotometric determination

We report on a combination of magnetic solid-phase extraction and spectrophotometric determination of bromate. Cetyltrimethylammonium ion was adsorbed on the surface of phenyl-functionalized silica-coated Fe₃O₄ nanoparticles (Ph-SiO₂@Fe₃O₄), and these materials served as the sorbent. The effects of surfactant and amount of sorbent, the composition of the desorption solution, the extraction time and temperature were optimized. Under optimized conditions, an enrichment factor of 12 was achieved, and the relative standard deviation is 2.9 % (for n?=?5). The calibration plot covers the 1–50 ng mL^?1 range with reasonable linearity (r ²?>?0.998); and the limit of detection is 0.5 ng mL^?1. The method is not interfered by ionic compounds commonly found in environmental water samples. It was successfully applied to the determination of bromate in spiked water samples.

Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk

To address food safety concerns of the public regarding the potential transfer of recombinant DNA (cry1Ab) and protein (Cry1Ab) into the milk of cows fed genetically modified maize (MON810), a highly specific and sensitive quantitative real-time PCR (qPCR) and an ELISA were developed for monitoring suspicious presence of novel DNA and Cry1Ab protein in bovine milk. The developed assays were validated according to the assay validation criteria specified in the European Commission Decision 2002/657/EC. The detection limit and detection capability of the qPCR and ELISA were 100 copies of cry1Ab μL^?1 milk and 0.4 ng mL^?1 Cry1Ab, respectively. Recovery rates of 84.9% (DNA) and 97% (protein) and low (<15%) imprecision revealed the reliable and accurate estimations. A specific qPCR amplification and use of a specific antibody in ELISA ascertained the high specificity of the assays. Using these assays for 90 milk samples collected from cows fed either transgenic (n?=?8) or non-transgenic (n?=?7) rations for 6 months, neither cry1Ab nor Cry1Ab protein were detected in any analyzed sample at the assay detection limits.

Voltammetric sensing of thallium at a carbon paste electrode modified with a crown ether

We describe a new method for differential-pulse anodic stripping voltammetric determination of thallium(I) using a carbon paste electrode modified with dicyclohexyl-18-crown-6. The effect of supporting electrolyte (type and pH), accumulation and reduction potential, and of time and amount of modifier were investigated by differential pulse anodic stripping voltammetry. A method was then worked out for the determination of thallium at low levels. Under optimized conditions, the response to Tl(I) is linear in the range from 3.0 to 250 ng mL^?1. The detection limit is 0.86 ng mL^?1. The sensor displays good repeatability (with a relative standard deviation of ±2.70 % for n?=?7) and was applied to the determination of Tl(I) in water, hair samples, and certified reference materials.

Enzyme-linked immunosorbent assay and immunochromatographic strip for rapid detection of atrazine in water samples

We have developed a heterologous direct competitive enzyme-linked immunosorbent assay (ELISA) and a colloidal gold-based immunochromatographic (ICG) strip for the determination of the herbicide atrazine in water samples. The ELISA had a half-maximum inhibition concentration (IC₅₀) of 0.12 ng mL^?1 and a limit of detection (LOD, calculated as the IC₁₅ value) of 0.01 ng mL^?1. The average of recoveries for all spiked water samples was 96.5%. There was a good correlation between the data determined by this ELISA and those obtained by high performance liquid chromatography (HPLC) (r ² ?=?0.996). The visual LOD of the ICG strip assay was 2 ng mL^?1. The assay process only took 10 min, and no sample pretreatment was required. Its high specificity, sensitivity and fast detection made the strip well suited for on-site screening of atrazine in water samples. Both the ELISA and the ICG strip assay are useful for rapid analysis of a large number of water samples at low cost.

Preconcentration of trace cadmium ion using magnetic graphene nanoparticles as an efficient adsorbent

Graphene-based magnetic nanoparticles (G-Fe₃O₄) were prepared and used as an effective adsorbent for the solid-phase extraction of trace quantities of cadmium from water and vegetable samples. The method avoids some of the time-consuming steps associated with traditional solid phase extraction. The excellent sorption property of the G-Fe₃O₄ system is attributed to π - π stacking interaction and hydrophobic interactions between graphene and the Cd-PAN complex. The effects of pH, the amount of G–Fe₃O₄, extraction time, type and volume of eluent, desorption time and interfering ions on the extraction efficiency were optimized. The preconcentration factor is 200. Cd(II) was then quantified by flame atomic absorption spectrometry with a detection limit of 0.32 ng mL^?1. The relative standard deviation (at 50 ng mL^?1; for n?=?10) is 2.45 %. The method has a linear analytical range from 1.1 to 150 ng mL^?1, and the recoveries in case of real samples are in the range between 93.1 % and 102.3 %.

Carbon paste electrode modified with a binuclear manganese complex as a sensitive voltammetric sensor for tryptophan

We report on a carbon paste electrode that was modified with a binuclear manganese(II) complex by the drop-coating method. A study on the mechanism of the electro-oxidation of tryptophan (Trp) at this electrode indicated that it enables Trp to be determined with good sensitivity and selectivity. Second-order derivative linear sweep voltammetry at pH 4.1 revealed that a sensitive anodic peak appears at 812?mV (vs. SCE) whose current is proportional to the concentration of Trp in the concentration range from 0.1 to 1.0???mol?L^?1 and 1.0 to 80???mol?L^?1, with a detection limit (S/N?=?3) of 0.08???mol?L^?1 (60?s of accumulation). The method was applied to the determination of Trp in amino acid injection solutions with satisfactory results.

A biopolymer-based carbon nanotube interface integrated with a redox shuttle and a D-sorbitol dehydrogenase for robust monitoring of D-sorbitol

We describe the preparation and characterization of a glassy carbon electrode modified with a bionanocomposite consisting of a hyaluronic acid, dispersed carbon nanotubes, and electrostatically bound toluidine blue. The electrode was used to detect NADH in the batch and flow-injection mode of operation. The electrode was further modified by immobilizing sorbitol dehydrogenase to result in biosensor for D-sorbitol that displays good operational stability, a sensitivity of 10.6???A?mM^?1?cm^?2, a response time of 16?s, and detection limit in the low micromolar range. The biosensor was successfully applied to off-line monitoring of D-sorbitol during its bioconversion into L-sorbose (a precursor in the synthesis of vitamin C) by Gluconobacter oxydans. The sample assay precision is 2.5% (an average RSD) and the throughput is 65?h^?1 if operated in the flow-injection mode. The validation of this biosensor against a reference HPLC method resulted in a slope of correlation of 1.021?±?0.001 (R ²?=?0.99997).

Label-free sandwich type of immunosensor for hepatitis C virus core antigen based on the use of gold nanoparticles on a nanostructured metal oxide surface

We report on the construction of a label-free electrochemical immunosensor for detecting the core antigen of the hepatitis C virus (HCV core antigen). A glassy carbon electrode (GCE) was modified with a nanocomposite made from gold nanoparticles, zirconia nanoparticles and chitosan, and prepared by in situ reduction. The zirconia nanoparticles were first dispersed in chitosan solution, and then AuNPs were prepared in situ on the ZrO₂-chitosan composite. In parallel, a nanocomposite was synthesized from AuNPs, silica nanoparticles and chitosan, and conjugated to a secondary antibody. The properties of the resulting nanocomposites were investigated by UV-visible photometry and transmission electron microscopy, and the stepwise assembly process was characterized by means of cyclic voltammetry and electrochemical impedance spectroscopy. An sandwich type of immunosensor was developed which displays high sensitivity to the HCV core antigen in the concentration range between 2 and 512?ng?mL^?1, with a detection limit of 0.17?ng?mL^?1 (at S/N?=?3). This immunosensor provides an alternative approach towards the diagnosis of HCV.

Anodic stripping voltammetry of silver(I) using a carbon paste electrode modified with multi-walled carbon nanotubes

We describe a silver(I)-selective carbon paste electrode modified with multi-walled carbon nanotubes and a silver-chelating Schiff base, and its electrochemical response to Ag(I). Effects of reduction potential and time, accumulation time, pH of the solution and the stripping medium were studied by differential pulse anodic stripping voltammetry and optimized. The findings resulted in a method for the determination of silver over a linear response range (from 0.5 to 235 ng?mL^?1) and with a detection limit as low as 0.08 ng?mL^?1. The sensor displays good repeatability (with the RSD of ±?2.75 % for 7 replicates) and was applied to the determination of Ag(I) in water samples and X-ray photographic films.

Electrochemical sensor for lead(II) ion using a carbon ionic-liquid electrode modified with a composite consisting of mesoporous carbon, an ionic liquid, and chitosan

A novel electrode was prepared that enables sensing of lead(II) ion. A suspension composed of ordered mesoporous carbon (OMC), an ionic liquid (IL), and chitosan was deposited on the highly conductive surface of a carbon ionic-liquid electrode (CILE). The surface of the sensing electrode was characterized by scanning electron microscopy and cyclic voltammetry. The new electrode can be used to determine lead(II) ion because the hydrophobic ionic liquid of the CILE can extract Pb(II), while the OMC accelerates the electron transfer rate between the electrode and Pb(II) and also strongly adsorbs Pb(II). The resulting electrode displays excellent and synergistic response to Pb(II) which is linear in the range from 0.05 to 1.4?μM, with a correlation coefficient of 0.997 and a detection limit of 25 nM.

Trans-membrane electron transfer in red blood cells immobilized in a chitosan film on a glassy carbon electrode

We have studied the trans-membrane electron transfer in human red blood cells (RBCs) immobilized in a chitosan film on a glassy carbon electrode (GCE). Electron transfer results from the presence of hemoglobin (Hb) in the RBCs. The electron transfer rate (k _s) of Hb in RBCs is 0.42 s^?1, and <1.13 s^?1 for Hb directly immobilized in the chitosan film. Only Hb molecules in RBCs that are closest to the plasma membrane and the surface of the electrode can undergo electron transfer to the electrode. The immobilized RBCs displayed sensitive electrocatalytic response to oxygen and hydrogen peroxide. It is believed that this cellular biosensor is of potential significance in studies on the physiological status of RBCs based on observing their electron transfer on the modified electrode.

A cloud point extraction for spectrophotometric determination of ultra- trace antimony without chelating agent in environmental and biological samples

We report on a simple, sensitive and reliable method for the cloud point extraction of antimony (Sb) and its subsequent spectrophotometric detection. It is based on the color reaction of Sb (III) with iodide in acidic medium and subsequent micelle-mediated extraction of tetraiodoantimonate using a non-ionic surfactant in the absence of any chelating agent. The effects of reaction and extraction parameters were optimized. The calibration plot is linear in the range of 0.80–95?ng?mL^?1 of antimony in the sample solution, with a regression coefficient (r) of 0.9994 (for n?=?9). The detection limit (at SNR?=?3) is 0.23?ng?mL^?1, and the relative standard deviations at 10 and 70?ng?mL^?1 of antimony are 3.32 and 1.85?% (at n?=?8), respectively. The method compared favorably to other methods and was applied to determine antimony in seawater, anti-leishmania drug (glucantime), and human serum.

Sensitive detection of enteropathogenic E. coli using a bfpA gene-based electrochemical sensor

We have developed a sensitive assay for enteropathogenic E. coli (EPEC) by integrating DNA extraction, specific polymerase chain reaction (PCR) and DNA detection using an electrode modified with the bundle-forming pilus (bfpA) structural gene. The PCR amplified products are captured on the electrode and hybridized with biotinylated detection probes to form a sandwich hybrid containing two biotinylated detection probes. The sandwich hybridization structure significantly combined the numerous streptavidin alkaline phosphatase on the electrode by biotin-streptavidin connectors. Electrochemical readout is based on dual signal amplification by both the sandwich hybridization structure and the enzyme. The electrode can satisfactorily discriminate complementary and mismatched oligonucleotides. Under optimal conditions, synthetic target DNA can be detected in the 1 pM to 10 nM concentration range, with a detection limit of 0.3 pM. EPEC can be quantified in the 10 to 10⁷ CFU mL^?1 levels within 3.5 h. The method also is believed to present a powerful platform for the screening of pathogenic microorganisms in clinical diagnostics, food safety and environmental monitoring.

Micro-solid phase extraction of ochratoxin A, and its determination in urine using capillary electrophoresis

We describe a simple, environmentally friendly and selective technique for the determination of ochratoxin A (OTA) in urine. It involves (a) the use of a molecularly imprinted polymer as a sorbent in micro-solid-phase extraction in which the sorbent is contained in a propylene membrane envelope, and (b) separation and detection by capillary electrophoresis (CE). Under optimized conditions, response is linear in the range between 50 and 300 ng mL^?1 (with a correlation coefficient of 0.9989), relative standard deviations range from 4 to 8 %, the detection limit for OTA in urine is 11.2 ng mL^?1 (with a quantification limits of 32.5 ng mL^?1) which is lower than those of previously reported methods for solid-phase extraction combined with CE. The recoveries of OTA from urine spiked at levels of 50, 150 and 300 ng mL^?1 ranged from 93 to 97 %.

Disposable immunoassay for hepatitis B surface antigen based on a graphene paste electrode functionalized with gold nanoparticles and a Nafion-cysteine conjugate

We report on the modification of a graphene paste electrode with gold nanoparticles (AuNPs) and a Nafion-L-cysteine composite film, and how this electrode can serve as a platform for the construction of a novel electrochemical immunosensor for the detection of hepatitis B surface antigen (HBsAg). To obtain the immunosensor, an antibody against HBsAg was immobilized on the surface of the electrode, and this process was followed by cyclic voltammetry and electrochemical impedance spectroscopy. The peak currents of a hexacyanoferrate redox system decreased on formation of the antibody-antigen complex on the surface of the electrode. Then increased electrochemical response is thought to result from a combination of beneficial effects including the biocompatibility and large surface area of the AuNPs, the high conductivity of the graphene paste electrode, the synergistic effects of composite film, and the increased quantity of HBsAb adsorbed on the electrode surface. The differential pulse voltammetric responses of the hexacyanoferrate redox pair are proportional to the concentration of HBsAg in the range from 0.5–800?ng?mL^?1, and the detection limit is 0.1?ng?mL^?1 (at an S/N of 3). The immunosensor is sensitive and stable.

Visual detection and microplate assay for Staphylococcus aureus based on aptamer recognition coupled to tyramine signal amplification

We have developed a specific method for the visual detection of Staphylococcus aureus based on aptamer recognition coupled to tyramine signal amplification technology. A biotinylated aptamer specific for S. aureus was immobilized on the surface of the wells of a microplate via biotin-avidin binding. Then, the target bacteria (S. aureus), the biotinylated-aptamer-streptavidin-HRP conjugates, biotinylated tyramine, hydrogen peroxide and streptavidin-HRP were successively placed in the wells of the microplate. After adding TMB reagent and stop solution, the intensity of the yellow reaction product can be visually inspected or measured with a plate reader. Under optimized conditions, there is a linear relationship between absorbance at 450 nm and the concentration of S. aureus in the 10 to 10⁷ cfu mL^?1 concentration range (with an R² of 0.9976). The limit of detection is 8 cfu mL^?1.

Ultrasensitive electrochemiluminescent immunoassay for morphine using a gold electrode modified with CdS quantum dots, polyamidoamine, and gold nanoparticles

We report on a novel electrochemiluminescent (ECL) immunoassay for the ultrasensitive determination of morphine by making use of a gold electrode which was modified with a nanocomposite film containing self-assembled polyamidoamine (PAMAM) CdS quantum dots and electrodeposited gold nanoparticles (Au-NPs). The highly uniform and well-dispersed quantum dots were capped with PAMAM dendrimers. Due to the synergistic effect of the modified quantum dots and the electrodeposited Au-NPs, the ECL response is dramatically enhanced. Under optimal experimental conditions, the immunoreaction between morphine and anti-morphine antibody resulted in a decrease of the ECL signal because of steric hindrance. The calibration plot is linear in the morphine concentration range from 0.2 to 180 ng?mL^?1, with a detection limit as low as 67 pg?mL^?1. The sensor was successfully applied to the determination of morphine in blood plasma. This kind of assay is expected to pave new avenues in label-free drug assays.

Electrochemical synthesis of a novel platinum nanostructure on a glassy carbon electrode, and its application to the electrooxidation of methanol

We show that the addition of white dextrin during the electrochemical deposition of platinum nanostructures (nano-Pt) on a glassy carbon electrode (GCE) results in an electrochemically active surface that is much larger than that of platinum microparticles prepared by the same procedure but in the absence of dextrin. The nano-Pt deposits are characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy, and electrochemical methods. The SEM images reveal deposits composed of mainly nanoparticles and short nanorods. The GCE was applied as a novel and cost-effective catalyst for methanol oxidation. The use of nano-Pt improves the electrocatalytic activity and the stability of the electrodes.

Preparation of Fe3O4@C@CNC multifunctional magnetic core/shell nanoparticles and their application in a signal-type flow-injection photoluminescence immunosensor

We describe here the preparation of carbon-coated Fe₃O₄ magnetic nanoparticles that were further fabricated into multifunctional core/shell nanoparticles (Fe₃O₄@C@CNCs) through a layer-by-layer self-assembly process of carbon nanocrystals (CNCs). The nanoparticles were applied in a photoluminescence (PL) immunosensor to detect the carcinoembryonic antigen (CEA), and CEA primary antibody was immobilized onto the surface of the nanoparticles. In addition, CEA secondary antibody and glucose oxidase were covalently bonded to silica nanoparticles. After stepwise immunoreactions, the immunoreagent was injected into the PL cell using a flow-injection PL system. When glucose was injected, hydrogen peroxide was obtained because of glucose oxidase catalysis and quenched the PL of the Fe₃O₄@C@CNC nanoparticles. The here proposed PL immunosensor allowed us to determine CEA concentrations in the 0.005–50 ng?·?mL^-1 concentration range, with a detection limit of 1.8 pg?·?mL^-1.