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 %.

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     

Determination of methylisothiocyanate in soil and water by HS-SPME followed by GC–MS–MS with a triple quadrupole

Methylisothiocyanate (MITC) is the main degradation product of metam sodium, a soil disinfectant widely used in agriculture, and is responsible for its disinfectant properties. Because MITC is highly toxic and volatile, metam sodium has to be applied in a manner that tries to reduce atmospheric emissions but still maintains adequate concentration of MITC in soil to ensure its disinfectant effect. Thus, monitoring of MITC concentrations in soil is required, and to this end sensitive, fast, and reliable analytical methods must be developed. In this work, a headspace solid-phase microextraction (HS-SPME) method was developed for MITC determination in water and soil samples using gas chromatography-tandem mass spectrometry (GC–MS–MS) with a triple-quadrupole analyzer. Two MS–MS transitions were acquired to ensure the reliable quantification and confirmation of the analyte. The method had linear behavior in the range tested (0.026–2.6 ng mL^?1 in water, 1–100 ng g^?1 in soil) with r ² over 0.999. Detection limits were 0.017 ng mL^?1 and 0.1 ng g^?1 in water and soil, respectively. Recoveries for five replicates were in the range 76–92 %, and RSD was below 7 % at the two spiking levels tested for each matrix (0.1 and 1 ng mL^?1 for water, 4 and 40 ng g^?1 for soil). The potential of using multiple HS-SPME for analyzing soil samples was also investigated, and its feasibility for quantification of MITC evaluated. The developed HS-SPME method was applied to soil samples from experimental plots treated with metam sodium following good agriculture practices. Figure

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Determination of fluoroquinolone antibiotics by microchip capillary electrophoresis along with time-resolved sensitized luminescence of their terbium(III) complexes

We report on the time-resolved detection of the three fluoroquinolone (FQs) antibiotics ciprofloxacin (CIP), enrofloxacin (ENR) and flumequine (FLU). On addition of terbium(III) ions, the terbium(III)-FQs chelates are formed in-situ in an on-capillary derivatization reaction of a microfluidic system. The laser-induced terbium(III)-sensitized luminescence of the chelates is measured at excitation/emission wavelengths of 337/545 nm. The analytes can be separated and quantified within less than 4 min. A solid phase extraction step for analyte preconcentration can be included prior to chelation and microchip capillary electrophoresis. The analytical ranges of the calibration graphs for CIP, ENR and FLU are from 10.6 to 60.0, 10.3 to 51.0, and 11.5 to 58.8 ng mL^?1, respectively, and the detection limits are 3.2, 3.1 and 3.6 ng mL^?1, respectively. The precision was established at two concentration levels of each analyte and revealed relative standard deviations in the range from 3.0 to 10.2 %. The method was applied to the analysis of FQ-spiked water samples. Figure

We report on the time-resolved detection of the three fluoroquinolone antibiotics. The analytes can be separated and quantified within less than 4 min. A solid phase extraction step for analyte preconcentration can be included prior to chelation and microchip capillary electrophoresis.     

Colloidal gold based immunochromatographic strip for the simple and sensitive determination of aflatoxin B1 and B2 in corn and rice

We have developed a simple and fast immunochromatographic test strip for the simultaneous quantitation of aflatoxin B₁ and aflatoxin B₂ in corn and rice. The strip contains three pads (sample, conjugate, and absorbing pad) and uses the respective polyclonal antibodies immobilized on gold nanoparticles. Matrix interferences were minimized by application of fugacity theory. Clean-up of samples and pre-treatment of strip pads is not required. The visual detection limit is 0.1 ng mL^?1, and the process can be completed within 5 min. Out of 113 natural samples, 16 rice and 27 corn samples (38% in total) were aflatoxin positive and the test results were confirmed by HPLC. The strip shows, however, high cross reactivity to aflatoxins G₁, G₂, and M₁. We consider this strip to possess wide applicability because of its ease of use, sensitivity, stability, and low cost.

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%.

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.

Chemiluminescent determination of the activity of caspase-3 using a specific peptide substrate and magnetic beads

We report on a new scheme for the determination of the activity of caspase-3 using a specific peptide labeled with N-(4-aminobutyl)-N-ethylisoluminol (ABEI) as a chemiluminescent (CL) probe and on the development of magnetic separation technology. Firstly, the ABEI-labeled and biotinylated peptide was prepared and conjugated to streptavidin-coated magnetic beads (MBs) to form f-MBs (functionalized magnetic beads). The f-MBs contain a site (DEVD, Asp-Glu-Val-Asp) that is cleaved by caspase-3. Upon cleavage, the terminal residue attached to ABEI can dissociate from the f-MBs and can be used for CL detection. CL intensity is linearly related to the concentration of caspase-3 in the range 1.0 to 600 ng mL^?1, with a detection limit of 0.3 ng mL^?1. The relative standard deviation of the assay is 3.6 % at a level of 50 ng mL^?1 of caspase-3 (for n?=?11). The CL assay has been applied to the determination of caspase-3 in Jurkat cell extract with recoveries between 96.6 % and 106.1 % (n?=?5).

Label-free microcantilever-based immunosensors for highly sensitive determination of avian influenza virus H9

We report on label-free immunosensors for the highly sensitive detection of avian influenza virus. The method makes use of the microcantilevers of an atomic force microscope onto which monoclonal antibodies against avian influenza virus were covalently immobilized. The factors influencing the performance of the resulting immunosensors were optimized by measuring the deflections of the cantilever via optical reflection, and this resulted in low detection limits and a wide analytical range. The differential deflection signals revealed specific antigen binding and their intensity is proportional to the logarithm of the concentrations of the virus in solution. Under optimal conditions, the immunosensors exhibit a linear response in the 7.6 ng mL^?1 to 76 μg mL^?1 concentration range of avian influenza virus, and the detection limit is 1.9 ng mL^?1.

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 %.

Triazine-modified magnetite nanoparticles as a novel sorbent for preconcentration of lead and cadmium ions

We report on a new sorbent for preconcentration of cadmium and lead ions that is based on triazine-functionalized magnetite nanoparticles that were prepared by direct silylation of magnetic nanoparticles with 3-aminopropyltriethoxysilane-2,4-bis(3,5-dimethylpyrazol)-triazine. The sorbent was characterized by IR spectroscopy, X-ray powder diffraction, scanning electron microscopy, thermal and elemental analysis. The sorbent was applied to the preconcentration of lead and cadmium ions which then were quantified by FAAS. The effects of sample pH value, extraction time, of type, concentration and volume of eluent, and of elution time were optimized. The limits of detection are 0.7 ng mL^?1 for Pb(II) ion and 0.01 ng mL^?1 for Cd(II). The effects of potentially interfering ions often found in real samples on the recovery in the determination of cadmium and lead ions in real samples were also investigated. The accuracy of the method was confirmed by analyzing the certified reference materials NIST 1571 (orchard leaves) and NIST 1572 (citrus leaves). Finally, the method was successfully applied to the determination of cadmium and lead ions in some fruit samples. Figure

We report on a new sorbent for preconcentration of cadmium and lead ions that is based on triazine-functionalized magnetite nanoparticles. After optimization of the preconcentration step the method was successfully applied to the determination of cadmium and lead ions in some fruit samples     

A novel test using dried blood spots for the chromatographic assay of methadone

A novel test has been developed for the analysis of methadone in dried blood spot specimens from patients undergoing methadone maintenance treatment. An isocratic reversed-phase high-performance liquid chromatography method with coulometric detection has been optimized for the determination of methadone. The clean-up of dried blood spots was performed by means of an original microextraction by packed sorbent procedure after microwave-assisted extraction of the drug with a suitable solvent. Extraction yields were satisfactory, always being higher than 90.0 %. The calibration curve was linear over the 4–500 ng mL^-1 concentration range. The method had satisfactory sensitivity (limit of quantitation of 4 ng mL^-1), precision (relative standard deviation less than 5.8 %), selectivity and accuracy (recovery greater than 87.0 %). It was successfully applied to dried blood spot samples collected from heroin-addicted patients undergoing methadone maintenance therapy at dosages between 40 and 240 mg day^-1. The statistical analysis (Bland–Altman plot) showed that the results were in good agreement with those found from the analysis of plasma samples obtained from the same patients. Thus, the method has proved to be suitable for the monitoring of methadone by means of dried blood spots.

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.

Molecularly imprinted solid-phase extraction monolithic capillary column for selective extraction and sensitive determination of safranine T in wolfberry

A method was developed to sensitively determine safranine T in wolfberry by molecularly imprinted solid-phase extraction (MISPE) coupled with high-performance liquid chromatography and laser-induced fluorescence detection (HPLC-LIF). The MISPE capillary monolithic column was prepared by water-bath in situ polymerization, using safranine T, methacrylic acid (MAA), and ethylene dimethacrylate (EDMA) as template, functional monomer, and cross-linker, respectively. The properties of the homemade MISPE capillary monolithic column, including capacity and specificity, were investigated under optimized conditions and the morphologies of inner polymers were characterized by scanning electron microscopy (SEM). The mean recoveries of safranine T in wolfberry ranged from 91.2 % to 92.9 % and the intraday and interday relative standard deviation (RSD) values all ranged from 3.4 % to 4.2 %. Good linearity was obtained over 0.001–1.0 μg mL^–1 (r?=?0.9999) with a detection limit (S/N?=?3) of 0.4 ng g^–1. Under the selected conditions, enrichment factors of over 90-fold were obtained and the extraction on the monolithic column effectively cleaned up the wolfberry matrix. The results demonstrated that the proposed MISPE-HPLC-LIF method could be applied to sensitively determine safranine T in wolfberry.

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.

Integration of lateral flow and microarray technologies for multiplex immunoassay: application to the determination of drugs of abuse

We report on a lateral flow microarray that combines multi-spot immunochip technology and immunochromatography. It can serve as a tool for the simultaneous detection of multiple analytes. The test zone of the nitrocellulose support comprises a microarray spotted with up to 32 antigens that can capture labeled gold-antibodies after lateral flow. The detection limits and detectable concentration ranges of the assay were characterized. The method was applied to the determination of drugs of abuse (and their metabolites) in urine, specifically of morphine, amphetamine, methamphetamine, and benzoylecgonine. The assay format is rapid (10 min), and has both a low relative standard deviation (< 9 %) and high recoveries (95–114 %). The detection limits (2–20 ng mL^–1 for drugs of abuse) are comparable to those of conventional single-analyte strip methods.

Solid phase extraction of heavy metal ions based on a novel functionalized magnetic multi-walled carbon nanotube composite with the aid of experimental design methodology

We report that magnetic multiwalled carbon nanotubes functionalized with 8-aminoquinoline can be applied to the preconcentration of Cd(II), Pb(II) and Ni(II) ions. The parameters affecting preconcentration were optimized by a Box-Behnken design through response surface methodology. Three variables (extraction time, magnetic sorbent amount, and pH value) were selected as the main factors affecting sorption, and four variables (type, volume and concentration of the eluent; elution time) were selected for optimizing elution. Following sorption and elution, the ions were quantified by FAAS. The LODs are 0.09, 0.72, and 1.0 ng mL^?1 for Cd(II), Ni(II), and Pb(II) ions, respectively. The relative standard deviations are <5.1 % for five separate batch determinations at 30 ng mL^?1 level of Cd(II), Ni(II), and Pb(II) ions. The sorption capacities (in mg g^?1) of this new sorbent are 201 for Cd(II), 150 for Pb(II), and 172 Ni(II). The composite was successfully applied to the rapid extraction of trace quantities of heavy metal ions in fish, sediment, soil, and water samples.

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.

Enantioselective Separation and Determination of Carnosine in Rat Plasma by Fluorescence LC for Stereoselective Pharmacokinetic Studies

A sensitive fluorescence liquid chromatographic analytical method was developed for the simultaneous determination of carnosine enantiomers in rat plasma. The method was applied to pharmacokinetic studies. Chiral separation of carnosine enantiomers was achieved by pre-column derivatization with o-phthaldialdehyde and the thiol N-acety-l-cysteine as derivating reagents. They were separated on an ODS column and detected by fluorescence detection (λ_ex = 350 nm, λ_em = 450 nm). γ-Aminobutyric acid was used as internal standard. The method was linear up to 6,000 ng mL^?1 for l-carnosine, 4,000 ng mL^?1 for d-carnosine. Low limit of quantitation (LLOQ) was 40 ng mL^?1 for each isomer. The relative standard deviations obtained for intra- and inter-day precision were lower than 12% and the recoveries were higher than 75% for both enantiomers. The method was applied to a stereoselective study on the pharmacokinetics of carnosine after oral administration with a single dose (carnosine, 75 mg kg^?1 for each isomer) to a rat. The initial data indicated that l-carnosine had a larger value of the highest plasma concentration than d-carnosine (C _max 5,344 vs. 1,914 ng mL^?1), and that of l-carnosine had a lower value of AUC_(0?∞) and t _1/2(h) (AUC_(0?∞) 5,306 vs. 6,321 ng h mL^?1, t _1/2 1.43 vs. 3.37 h). Our results indicated that the pharmacokinetic of l-carnosine and d-carnosine revealed enantioselective properties significantly.     

Solid phase ligand-less extraction of cadmium(II) using a silica gel modified with an amino-functionalized ionic liquid

A method was developed for the determination of cadmium(II) by ligand-less solid phase extraction that is based on the direct retention of Cd(II) in a mini-column filled with a silica gel modified with an amino-functionalized ionic liquid. The effects of pH, sample volume and its flow rate, eluent concentration and its volume, the flow rate of eluent, and of potential interferences on extraction and desorption were optimized. Following its determination by electrothermal atomic absorption spectrometry, the detection limit for Cd(II) is 8.9 ng L^?1, and the relative standard deviation is 2.3 % (at 1.0 ng mL^?1; for n?=?5). The method was applied to the analysis of Cd(II) in a certified reference material (laver; GBW10023), and the recoveries ranged from 97.0 to104.0 %