Plastic disposable choline biosensors based on ruthenized-carbon screen-printed electrodes were prepared and their use for monitoring organophosphorus pesticides and carbamates is described. The presence of 0.5% ruthenium on activated carbon mixed to form a simple graphite-based ink for the working electrode surface increased the sensitivity towards hydrogen peroxide. The choline biosensor is based on such an electrode coupled with choline oxidase immobilized by adsorption and was used to detect the inhibition effect of carbamates and organophosphorus pesticides on acetylcholinesterase. With the optimized procedure described (pH, buffer composition, incubation time, substrate concentration), concentrations of pesticides (Carbofuran) as low as 1 nM could be detected.
We describe a method for magnetic solid phase extraction of trace-levels of Hg(II) ions by using Fe3O4 nanoparticles (NPs) covered with a shell of silica and modified with the chelator N-(2-acetylaminoethyl)-N-(3-triethoxysilylpropyl)thiourea. The new magnetic NPs enable rapid magnetic separation, thus leading to higher efficiency and accuracy. The extracted Hg(II) ions on the NPs were directly quantified using a mercury analyzer. Possible interferents are widely eliminated in this highly selective extraction process, and the NPs are not exerting an interfering effect either. The method has an enrichment factor of 100, and extraction recoveries are between 95 and 107 % when using 10 mg of the extracting NPs. The method works over a wide range of pH values and can be applied to even complex natural samples. The effects of pH value, extraction time, sample volume and adsorbent amount on the extraction efficiency were optimized. Under the optimal conditions, the detection limit is as low as 17 ng L-1. The method was applied to the preconcentration and detection of Hg(II) in three natural water samples using the standard addition method.
The characterisation of disposable screen-printed electrodes for stripping analysis is described. The graphite surface of the working electrode is used as substrate for plating a thin mercury film, which allows the electrochemical preconcentration of heavy metals. Optimisation procedures and experimental results are presented. Detection limits around the ppb level were obtained for different metals [Pb(II), Cd(II), Cu(II)].
Two different types of w-substituted alkanethiol/disulfide compounds have been used to prepare monolayer architectures on gold serving as platforms for the immobilization of receptor probe molecules - antibodies. These are: (i) carboxylic acid alkanethiols post-reacted with amino biotin to generate streptavidin surfaces, and (ii) N-hydroxysuccinimide-terminated disulfide surfaces. The properties of the monolayers, with and without attached receptor probe molecules, were analysed using infrared spectroscopy, ellipsometry, fluorescence scanning and atomic force microscopy. Several experimental parameters, such as condensation reagents, additives, probe and target concentrations and immobilization time, were systematically varied to determine the dynamic range and to optimize the sensitivity and signal-to-noise ratio of the biochip platforms. Fluorescence screening using Cy5-labelled antigens finally demonstrated that both surfaces could be successfully employed to immobilize the antibodies. The pros and cons of the two approaches are also discussed.
We report on an indirect optical method for the determination of glucose via the detection of hydrogen peroxide (H2O2) that is generated during the glucose oxidase (GOx) catalyzed oxidation of glucose. It is based on the finding that the ultraviolet (~374 nm) and visible (~525 nm) photoluminescence of pristine zinc oxide (ZnO) nanoparticles strongly depends on the concentration of H2O2 in water solution. Photoluminescence is quenched by up to 90 % at a 100 mM level of H2O2. The sensor constructed by immobilizing GOx on ZnO nanoparticles enabled glucose to be continuously monitored in the 10 mM to 130 mM concentration range, and the limit of detection is 10 mM. This enzymatic sensing scheme is supposed to be applicable to monitoring glucose in the food, beverage and fermentation industries. It has a wide scope in that it may be extended to numerous other substrate or enzyme activity assays based on the formation of H2O2, and of assays based on the consumption of H2O2 by peroxidases.
This review examines basic principles and applications of voltammetric electronic tongues. It is introduced by a description of the concept of electronic tongues or taste sensors followed by a general overview of electrochemical measurement principles that have been used for electronic tongues. A special emphasis is given on measurement principles for voltammetric electronic tongues, also including pulse voltammetry and variable reduction. Applications of voltammetric electronic tongue are described, such as in the food industry, environmental analysis, paper and pulp industry, household appliances and agriculture. Future developments of the concept, such as self polishing or miniaturized devices are also described. Finally, a continuous measurement system for chemical oxygen demand (COD), which has been commercialized, is depicted.
Tetraoctadecylammonium bromide (TOAB, (CH3(CH2)17)4N+Br–) has been used to print temporary hydrophobic barriers on carboxymethylated dextran (CMD) hydrogels to create a generic platform for protein microarray applications. The primary reason for printing temporary hydrophobic barriers is to prevent cross-contamination and overflow during microdrop dispensing. Equally important is to eliminate the risk for non-specific binding to the barriers during analyte exposure. This has been accomplished by introducing a regeneration step that removes the barriers after ligand immobilization. The overall fabrication process was characterized by microscopic wetting, atomic force microscopy, imaging ellipsometry, fluorescence microscopy, surface plasmon microscopy and biospecific interaction analysis. A series of model proteins including transferrin, Protein A, anti-myoglobin and bovine serum albumin was spotted into the TOAB-defined areas under different experimental conditions, e.g. at increased humidity and reduced substrate temperature or with glycerol as an additive in the protein solution. Much emphasis was devoted to studies aiming at exploring the homogeneity and activity of the immobilized proteins. The printed barriers were removed after protein immobilization using tert-n-butyl alcohol (TBA). TBA was found to be a very efficient agent as compared to previously used salt regeneration solutions, and the regeneration time could be reduced from 30 to 10 minutes. Finally, the potential of using the well established CMD hydrogel chemistry as a platform for protein microarrays was exploited using surface plasmon microscopy.
Microcontact printing (µCP) has been used to introduce temporary hydrophobic barriers on carboxymethylated dextran (CMD) hydrogels on gold. Among the investigated types of inks, tetraoctadecylammonium bromide (TOAB), electrostatically bound to the CMD layer, provided the most well-defined features both with respect to pattern-definition and reversibility upon exposure to a regeneration solution. The printed patterns were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), microscopic wetting and imaging null ellipsometry to explore the influence of concentration of ink solution and contact time on the appearance of the printed layer. AFM revealed that the printed TOAB molecules aggregated into clusters rather than into a homogeneous mono- or multilayer on the CMD hydrogel. It was also observed that printed areas of TOAB that are larger than 25?µm are inhomogeneous most likely because of an edge transfer lithography (ETL) mechanism. A protein model system based on Protein A-rabbit antimouse Fc ? was used to evaluate the potential of the patterned surface as a protein microarray chip by means of surface plasmon microscopy (SPM). Moreover, non-specific adsorption of several proteins onto TOAB barriers was also studied using surface plasmon resonance (SPR), and it is evident that undesired adsorption can be eliminated by removing barriers after ligand immobilization, but prior to analyte exposure, by treating the patterned surface with a simple salt regeneration solution. © Springer-Verlag/Wien 2004.
The authors describe an amperometric biosensor for the amino acid L-arginine (L-Arg). It is based on the use of a Nafion/Polyaniline (PANi) composite on a platinum screen-printed electrode (Pt-SPE) using a novel recombinant arginine deiminase isolated from Mycoplasma hominis. The protein was over-expressed, purified and employed as a biorecognition element of the sensor. Enzymatic hydrolysis of L-Arg leads to the formation of ammonium ions which diffuse into the Nafion/PANi layer and induce the electroreduction of PANi at a potential of -0.35 V (vs Ag/AgCl). L-Arg sensitivity is 684 +/- 32 A.M-1.m(-2), and the apparent Michaelis-Menten constant K-M(app)) is 0.31 +/- 0.05 mM. The calibration plot is linear over the range 3-200 mu M L-Arg, the limit of detection is 1 mu M, and the response time (for 90% of the total signal change to occur) is 15 s. The sensor is selective and exhibits good storage stability (amp;gt; 1 month without loss in signal). The biosensor was applied to the analysis of L-Arg in pharmaceutical samples and of ammonium and L-Arg in spiked human plasma obtained from blood of healthy volunteers and those with a hepatic disorder. Data generated were found to be in good agreement with a reference fluorometric enzymatic assay.
A novel electrocatalytic material for oxygen reduction, based on polyaniline in combinationwith copper, was developed and utilised for the direct voltammetric quantification of ammonium ions. Consecutive electrode modification by electrodeposited copper, a Nafion membrane and electropolymerised polyaniline resulted in an electrocatalytic composite material which the retained conductivity at neutral pH. Ammonia complex formation with Cu (I) caused the appearance of oxygen electrocatalysis, which was observed as an increase in cathodic current. This Faradaic phenomenon offered the advantage of direct voltammetric detection and was utilised for ammonium electroanalysis. The developed quantification protocol was applied for ammonium assay in human serum and compared with the routine approach for clinical analysis.