Development of a stable biosensor based on a SiO2 nanosheet–Nafion–modified glassy carbon electrode for sensitive detection of pesticides

SiO₂ nanosheets (SNS) have been prepared by a chemical method using montmorillonite as raw material and were characterized by scanning electron microscopy and X-ray diffraction. SiO₂ nanosheet–Nafion nanocomposites with excellent conductivity, catalytic activity, and biocompatibility provided an extremely hydrophilic surface for biomolecule adhesion. Chitosan was used as a cross-linker to immobilize acetylcholinesterase (AChE), and Nafion was used as a protective membrane to efficiently improve the stability of the AChE biosensor. The AChE biosensor showed favorable affinity for acetylthiocholine chloride and catalyzed the hydrolysis of acetylthiocholine chloride with an apparent Michaelis–Menten constant of 134 μM to form thiocholine, which was then oxidized to produce a detectable and fast response. Based on the inhibition by pesticides of the enzymatic activity of AChE, detection of the amperometric response from thiocholine on the biosensor is a simple and effective way to biomonitor exposure to pesticides. Under optimum conditions, the biosensor detected methyl parathion, chlorpyrifos, and carbofuran at concentrations ranging from 1.0?×?10^?12 to 1?×?10^?10?M and from 1.0?×?10^?10 to 1?×?10^?8?M. The detection limits for methyl parathion, chlorpyrifos, and carbofuran were 5?×?10^?13?M. The biosensor developed exhibited good sensitivity, stability, reproducibility, and low cost, thus providing a new promising tool for analysis of enzyme inhibitors.

Mixed immunoassay design for multiple chemical residues detection

In this research, a mixed immunoassay design for multiple chemical residues detection based on combined reverse competitive enzyme-linked immunosorbent assay (ELISA) procedure was developed. This method integrated two reverse ELISA reactions in one assay by labeling horseradish peroxidase to deoxynivalenol (DON) and orbifloxacin. Within this method, IC50 of the two mAbs for each analyte we produced ranged from 23?～?68 ng?mL^?1 for DONs and 4.1?～?49 ng?mL^?1 for quinolones (QNs). The limit of detection measured by IC10 was achieved at 0.45–1.3 ng?mL^?1 for DONs and 0.59–6.9 ng?mL^?1 for QNs, which was lower than the maximum residue levels. Recoveries in negative samples spiked at concentrations of 100, 200, and 500 ng?mL^?1 ranged from 91.3 to 102.2 % for DONs and 88.7–98.05 % for QNs with relative standard deviation less than 9.88 and 12.67 %. The results demonstrated that this developed immunoassay was suitable for screening of low molecular weight contaminants.

In vitro monitoring of picogram levels of risperidone in human urine via luminollysozyme flow injection chemiluminescence

The anti-schizophrenic drug risperidone (RSP) exerts an inhibitory effect on the chemiluminescence (CL) of the luminol-lysozyme system. This finding forms the basis for a sensitive flow injection method for its determination at picogram levels. RSP binds to Trp62 in the lysozyme, and this leads to a conformational change upon which the CL of the system is quenched. The decrease in CL is proportional to the logarithm of the concentration of RSP, and the calibration graph is linear in the range from 0.1 pg?mL^?1 to 1.0 ng?mL^?1, with relative standard deviations of <5.0%, and a detection limit of 0.05 pg?mL^?1 (3σ). At a flow rate of 2.0 mL?min^?1, the whole process including sampling and washing is completed within 20 s. The method was successfully applied to monitoring RSP in human urine after incorporation of 2 mg of RSP, with a total excretion of 16.6% within 8.5 h.

Analysis of dextromethorphan and pseudoephedrine in human plasma and urine samples using hollow fiber-based liquid–liquid–liquid microextraction and corona discharge ion mobility spectrometry

We report on the use of hollow fiber liquid-liquid-liquid microextraction (HF-LLLME) followed by corona discharge ion mobility spectrometry for the determination of dextromethorphan and pseudoephedrine in urine and plasma samples. The effects of pH of the donor phase, stirring rate, ionic strength and extraction time on HF-LLLME were optimized. Under the optimized conditions, the linear range of the calibration curves for dextromethorphan in plasma and urine, respectively, are from 1.5 to 150 and from 1 to 100 ng mL^?1. The ranges for pseudoephedrine, in turn, are from 30 to 300 and from 20 to 200 ng mL^?1. Correlation coefficients are better than 0.9903. The limits of detection are 0.6 and 0.3 ng mL^?1 for dextromethorphan, and 8.6 and 4.2 ng mL^?1 for pseudoephedrine in plasma and urine samples, respectively. The relative standard deviations range from 6 to 8%.

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.

A strategy to enhance the thermal stability of a nanostructured polypyrrole-based coating for solid phase microextraction

We report on a nanostructured self-doped polypyrrole (SPPy) film that was prepared by an electrochemical technique in an electrolyte containing fluorosulfonic acid as the sulfonation reagent. The film was applied as a new fiber material for solid-phase microextraction (SPME) of the pesticides lindane, heptachlor, aldrin, endosulfans I and II prior to their quantitation by GC with electron capture detection. The SPPy nanoparticles have a diameter of <100?nm. The introduction of covalently bound sulfo groups into the backbone of the polymer resulted in improved temperature resistance (~350?°C) and satisfactory extraction efficiency. The thermal stability of the SPPy fiber is superior to common polypyrrole fibers. Extraction was optimized by means of the Taguchi orthogonal array experimental design with an OA₁₆ (4⁵) matrix including extraction temperature, extraction time, salt concentration, stirring rate, and headspace volume. The method displays good repeatability (RSD?<?6%) and linearity (in the range from 0.78 to 100?ng?mL^?1; with an R² of >0.998. The detection limits are <0.23?ng?mL^?1. The method was successfully applied to the analysis of the pesticides in skimmed milk and fruit juice samples, and recoveries are from 84?±?1 to 105?±?1%.

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 %.

Determination of organophosphate pesticides using an acetylcholinesterase-based biosensor based on a boron-doped diamond electrode modified with gold nanoparticles and carbon spheres

We report on a biosensor for organophosphate pesticides (OPs) by exploiting their inhibitory effect on the activity of acetylcholinesterase (AChE). A boron-doped diamond (BDD) electrode was modified with a nanocomposite prepared from carbon spheres (CSs; with an average diameter of 500 nm) that were synthesized from resorcinol and formaldehyde, and then were coated with gold nanoparticles (AuNPs) by chemically growing them of the CSs. Compared to a bare BDD electrode, the electron transfer resistance is lower on this new electrode. Compared to an electrode without Au-NPs, the peak potential is negatively shifted by 42 mV, and the peak current is increased by 55 %. This is ascribed to the larger surface in the AuNP-CS nanocomposite which improves the adsorption of AChE, enhances its activity, and facilitates electrocatalysis. Under optimum conditions, the inhibitory effect of chlorpyrifos is linearly related to the negative log of its concentration in the 10^?11 to 10^?7 M range, with a detection limit of 1.3?×?10^?13 M. For methyl parathion, the inhibition effect is linear in the 10^?12 to 10^?6 M range, and the detection limit is 4.9?×?10^?13 M. The biosensor exhibits good precision and acceptable operational and temporal stability.

Simultaneous determination of carbazole-based explosives in environmental waters by dispersive liquid—liquid microextraction coupled to HPLC with UV-Vis detection

Dispersive liquid-liquid microextraction as a rapid, simple and efficient method coupled with high performance liquid chromatography-UV-Vis detection was used for sample preparation and subsequent determination of carbazole, tri nitro carbazole (TrNC) and tetra nitro carbazole in water samples. The influence of several important variables on the extraction efficiency has been evaluated. The methods works best with chloroform as an extractant and acetonitrile as the dispersive solvent. Under optimum conditions, the calibration curve is linear in the range from 0.007 to 1.75?μg?mL^?1 for TNC, 0.006 to 1.52?μg?mL^?1 for TrNC, and 0.008–2.10?μg?mL^?1 for carbazole. The limits of detection (LODs; at a signal-to-noise ratio of 3), range from 1.7 to 1.1?ng?mL^?1, for TNC, TrNC and carbazole. Also, the relative standard deviations (RSD, n?=?6) for the extraction of TNC (at 174?ng?mL^?1), TrNC (at 151?ng?mL^?1) and carbazole (at 84?ng?mL^?1) vary between 4.1 and 5.2%. The enrichment factors range from 179 to 186. The method was successfully applied to the determination of TNC, TrNC and carbazole in environmental samples.

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.

Dispersive liquid-liquid microextraction of phenolic compounds using solidified floating organic droplets, and their determination by HPLC

We have developed a simple and efficient method for dispersive liquid-liquid microextraction of 4-nitrophenol, 2-naphthol and bisphenol A in real water samples. It is making use of solidified floating organic droplets of 1-dodecanol which has low density and a proper melting point. The type and volume of extraction solvent and dispersive solvent, the effect of salts, pH value and extraction time were optimized and resulted in enrichment factors of 84 for 4-nitrophenol, 123 for 2-naphthol, and 97 for bisphenol A. The limits of detection by HPLC are 1.50, 0.10 and 1.02 ng · mL^?1, respectively. Excellent linearity is observed in the concentration range from 10 to 800 ng · mL^?1, with coefficients of correlation ranging from 0.9988 to 0.9999. The relative standard deviations (for n?=?5) are from 3.2 to 5.3 %, and relative recoveries for the three phenols in tap, river and spring water range from 85.0 to 105.0 %, 98.3 to 110.0 %, and 98.6 to 109.0 %, respectively.

New method for microextraction of ultra trace quantities of gold in real samples using ultrasound-assisted emulsification of solidified floating organic drops

A method was developed for the determination of gold ion in water samples using microextraction based on the ultrasound-assisted emulsification of solidified floating organic drops, followed by the flame atomic absorption spectrometry. N-(4-{4-[(anilinocarbothioyl)amino]benzyl}phenyl)-N-phenylthiourea was used as chelating agent. The parameters affecting the extraction and complex formation (including the type and volume of the extracting solvent, time of sonication and centrifugation, pH, amount of the chelating agent, and sample ionic strength) were optimized. Under the optimum conditions, the calibration graph is linear in the range from 1.5 to 400 ng mL^?1, with a limit of detection of 0.45 ng mL^?1. The relative standard deviation for ten replicate determinations of gold ion in a concentration of 175 ng mL^?1 was 1.7%. The procedure was successfully applied to the determination of gold in water samples, in pharmaceutical and synthetic samples, and in a standard reference material.

Highly sensitive electrochemical immunoassay for zearalenone in grain and grain-based food

We have developed a highly sensitive electrochemical immunoassay for the quantitation of zearalenone (ZEN), a mycotoxin produced by Fusarium species. In this enzyme linked immunosorbent assay, the enzymatic conversion of the substrate p-nitrophenylphosphate is detected by a microplate reader and the signal subsequently converted into an electrical signal. The concentrations of coating antigen (ZEN-ovalbumin), of monoclonal antibody, and of goat anti-mouse antibody labeled with alkaline phosphatase were optimized. In terms of electrochemical detection, the types and pH values of the buffers, the conditions for agitating, and scanning frequency were optimized. The effective detection range of this immunoassay is quite wide (0.004 to 9.5 ng?mL^?1), and the limit of detection is 2 pg?mL^?1. ZEN-free corn, wheat, and grain-based food samples were spiked with ZEN and analyzed by this method, and recoveries were found to range from 91.6 % to 113.0 %. Unlike previously described electrochemical methods, this method is both highly sensitive and has a wide working range. The method is fast and thus provides a platform for high-throughput analysis that meets the current need to monitor trace levels of analytes in grain and grain-based food.

Development of a haemolytic–enzymatic assay with mediated amperometric detection for palytoxin analysis: application to mussels

An electrochemical sensor for palytoxin (PlTX) detection, based on a strip of eight screen-printed electrodes connected to a cost-effective and portable apparatus, is reported. Sheep erythrocytes were used to test the palytoxin detector and degree of haemolysis was evaluated by measuring release of the cytosolic lactate dehydrogenase (LDH). Percentage haemolysis and, therefore, the amount of LDH measured, by use of NADH/pyruvate and appropriate electrochemical mediators, was correlated with the concentration of the toxin. Two different electrochemical approaches were investigated for evaluation of LDH release, but only one based on the use of a binary redox mediator sequence (phenazine methosulfate in conjugation with hexacyanoferrate(III)) proved useful for our purpose. After analytical and biochemical characterization, the sensor strip was used to measure palytoxin. Sheep blood and standard solutions of PlTX were left to react for two different incubation times (24 h or 4 h), resulting in working ranges of 7?×?10^?3–0.02 ng mL^?1 and 0.16–1.3 ng mL^?1, respectively. The specificity of the test for palytoxin was evaluated by use of ouabain, which acts in the same way as PlTX on the Na⁺/K⁺-ATPase pump. A cross-reactivity study, using high concentrations of other marine biotoxins was also conducted. Experiments to evaluate the matrix effect and recovery from mussels are discussed.

Selective light-triggered chemiluminescence between fluorescent dyes and luminol, and its analytical application

We report herein a novel chemiluminescence (CL) phenomenon triggered by light irradiation when a fluorescent dye, for example hematoporphyrin, fluorescein, eosin, or methylene blue is present in the luminol solution. A possible mechanism is proposed for the photoinduced chemiluminescence (PICL) reaction. Compared with reported methods for CL triggering, for example flow-injection, static reagent injection, and the electrochemical technique, the proposed in-situ PICL method presented has three advantages. First, the method is more selective, because the PICL signal of the target fluorescent dyes is initiated by excitation at a selective wavelength only. Second, the space and time resolution of the PICL method are better. Last, and most important, compared with injecting a reagent or inserting a electrode into the CL system to initiate the CL reaction, with the in-situ PICL method there is no physical interference with the target detecting system. All these advantages of the PICL method indicate it has many potential applications in the analytical sciences. The proposed method was applied to analysis of urine containing adrenaline. The linear range for adrenaline is 2.0?×?10^?10–1.0?×?10^?7 g mL^?1 and the detection limit is 6.0?×?10^?11 g mL^?1.

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.

Use of gold nanoparticle-labeled secondary antibodies to improve the sensitivity of an immunochromatographic assay for aflatoxin B1

We describe a sensitive method for the immunochromatographic determination of aflatoxin B1. It is based on the following steps: 1) Competitive interaction between non-labeled specific primary antibodies and target antigens in a sample and in the test zone of a membrane; 2) detection of the immune complexes on the membrane by using a secondary antibodies labeled with gold nanoparticles. The method enables precise adjustment of the required quantities of specific antibodies and the colloidal (gold) marker. It was applied in a lateral flow format to the detection of aflatoxin B1 and exhibits a limit of detection (LOD) of 160 pg?·?mL^?1 if detected visually, and of 30 pg?·?mL^?1 via instrumental detection. This is significantly lower than the LOD of 2 ng?·?mL^?1 achieved by conventional lateral flow analysis using the same reagents. Figure

Immunochromatography with secondary labeled antibodies caused 10-fold decrease of detection limit     

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.

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.

Magnetic titanium oxide nanoparticles for hemimicelle extraction and HPLC determination of organophosphorus pesticides in environmental water

We report on a method for the extraction of organophosphorus pesticides (OPPs) from water samples using mixed hemimicelles and magnetic titanium dioxide nanoparticles (Fe₃O₄@TiO₂) modified by cetyltrimethylammonium. Fe₃O₄@TiO₂ nanoparticles were synthesized by a hydrothermal process and then characterized by scanning electron microscopy and Fourier transform IR spectrometry. The effects of the quantity of surfactant, extraction time, desorption solvent, pH value, extraction volume and reuse of the sorbent were optimized with respect to the extraction of OPPs including chlorpyrifos, dimethoate, and trichlorfon. The extraction method was applied to analyze OPPs in environmental water using HPLC along with UV detection. The method has a wide linear range (100–15,000 ng L^?1), good linearity (r?>?0.999), and low detection limits (26–30 ng L^?1). The enrichment factor is ~1,000. The recoveries (at spiked levels of 100, 1,000 and 10,000 ng L^?1) are in the range of 88.5–96.7 %, and the relative standard deviations range from 2.4 % to 8.7 %.