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At least 937 records · Page 52Linked to original sources
PubMed1989

Pulsed amperometric detection of glucose in biological fluids at a surface-modified gold electrode.

A nonenzymatic glucose sensor that utilizes permselective membranes to achieve the selectivity required for screening glucose in biological fluids has been described. Interference from endogenous oxidizable substances such as amino acids, urea, ascorbic acid, and uric acid, as well as the effect of chloride and proteins on glucose response, is studied by using flow injection analysis. A set of membranes made of Naflin perfluorinated membrane and collagen, when arranged in front of the working electrode (gold), result in significant improvement in the system selectivity. Even at physiological pH, which is far from being the optimum pH for pulsed amperometric detection of carbohydrates, the sensor shows a good limit of detection (4-5 micrograms of glucose injected).

Acetaminophen↗
PubMed1990

Complete on-line determination of biopolymer molecular weight via high-performance liquid chromatography coupled to low-angle laser light scattering, ultraviolet, and differential refractive index detection.

An optically modified high-performance liquid chromatography refractive index detector was developed to allow complete on-line determinations for biopolymer molecular weights. On-line concentration, refractive index, specific refractive index increment (dn/dc2)mu, and Rayleigh factor were determined under flow injection analysis (FIA) and size exclusion chromatography (SEC) conditions using low-angle laser light scattering, ultraviolet, and modified refractive index detection. This instrumental system is capable of determining absolute on-line molecular weights. The error and time requirements involved in conventional methodologies for proteins have been reduced. Sample quantities have been reduced from 150 to 200 mg, in conventional off-line methods, to less than 2 mg for on-line FIA and 0.5 mg for on-line SEC, if mass absorptivities (a) are known. Otherwise, the determination of a will be the most sample-demanding step, requiring about 3 mg of the pure protein. On-line measurements of (dn/dc2)mu are in good agreement with traditional off-line values established at Donnan equilibrium (usually within 5%). In addition, this technique provides true injected mass as determined by the UV detector, after chromatographic exposure where losses may occur, which is then used in the calculation of biopolymer molecular weight.

Calibration↗
PubMed1990

Liposome flow injection immunoassay: implications for sensitivity, dynamic range, and antibody regeneration.

We have developed a liposome-based flow injection immunoassay (FIIA) system for quantitation of a clinical analyte, theophylline. With very minor changes in assay format, this procedure can also be used for the quantitation of anti-theophylline. Automated sequential analyses were performed at room temperature with picomole sensitivity and a day-to-day coefficient of variation of less than 5% for aqueous solutions. The system components include liposomes that contain fluorophores in their aqueous centers and an immobilized-antibody reactor column. The immunoreactor was regenerated hundreds of times over 3 months of continuous use with no measurable loss of antibody activity. The two assay formats studied produced distinct dynamic ranges for their respective analytes. The special advantages of using flow injection analysis for immunoassays and of using liposomes in FIIA are discussed.

Antibodies↗
PubMed2001

Nonaqueous catalytic fluorometric trace determination of vanadium based on the pyronine B-hydrogen peroxide reaction and flow injection after cloud point extraction.

The catalytic effect of vanadium on the pyronine B-H2O2 system is examined. Enhancement of the catalytic reaction rate along with the efficiency and selectivity against vanadium is achieved in a formic acid environment in the presence of a nonionic surfactant (Triton X-114). Elimination of drastic interference caused by inorganic acids and aqueous matrix along with a 50-fold preconcentration of vanadium are facilitated through cloud point extraction of its neutral complex with 8-quinolinol in an acidic solution. Subsequent flow injection analysis (FIA) with fluorometric detection renders the proposed method ideal for selective and cost-effective determination of as little as 0.020 microng L(-1) vanadium in environmental, biological, and food substrates. The preconcentration step can be applied simultaneously to multiple samples, allowing for massive preparation prior to analysis, compensating, thus, for the time-consuming procedure.

Catalysis↗
PubMed2002

A flow method with photocatalytic oxidation of dissolved organic matter using a solid-phase (TiO2) reactor followed by amperometric detection of consumed oxygen.

A photocatalytic sensor for the determination of chemical oxygen demand (COD) using titanium dioxide, based on the use of a pair of oxygen electrodes and flow injection analysis, is described. The measuring principle is based on the direct determination of the oxygen concentration change resulting from photocataltic oxidation of organic compounds. One of the two oxygen electrodes, the reference oxygen electrode, was utilized to measure the reference signal responding only to oxygen present in the injected samples. Oxygen consumption due to the TiO2-catalyzed photochemical oxidation of organic compounds in samples was monitored with the working oxygen electrode. The COD value of this sensor was calculated as the difference of the currents at reference and working oxygen electrodes, respectively. The operation characteristics of the sensor are demonstrated using artificially treated wastewater as a substrate. The sensor was also applied to the determination of COD in real water samples from dam reservoirs (n = 20) all over Japan. The results were in good agreement with those from the conventional COD methods (i.e., permanganate and dichromate methods).

Electrochemistry↗
PubMed2002

Simultaneous multiple substrate tag detection with ESI-ion trap MS for in vivo bacterial enzyme activity profiling.

A bacterial identification method in which multiple enzyme activities are measured simultaneously and in vivo with electrospray ionization-mass spectrometry (ESI-MS) is described. Whole-cell bacteria are immobilized onto a filter support and incubated with a mixture of substrates. Each substrate is chosen to measure a specific enzyme activity of a targeted bacterium and to produce a tag of unique molecular weight. After a predetermined incubation time, the solution is filtered, and the supernatant consisting of a mixture of released tags and unhydrolyzed substrates is directly analyzed, without chromatographic separation, by ESI-MS. Bacteria remain viable on the filter for further analyses. The method was tested by measuring the aminopeptidase activity of the bacteria Escherichia coli, Bacillus subtilis, Bacillus cereus, and Pseudomonas aeruginosa. The resulting aminopeptidase enzyme profiles allowed the differentiation between the four bacteria tested. The method is rapid, since a multiplex advantage is realized when assaying for multiple enzymes, and it is amenable to automation via a flow injection analysis setup.

Aminopeptidases↗
PubMed2002

Electrochemically controlled solid-phase microextraction based on conductive polypyrrole films.

Solid-phase microextraction (SPME) fiber coatings based on conductive polypyrrole films were prepared for the electrochemical extraction and desorption of ionic analytes. Simple preparation of each of the PPY extraction coatings on a platinum wire was possible with a constant potential method, but more importantly, cycling of the film between oxidation and reduction potentials facilitated the extraction and desorption of ionic analytes. The analytes were desorbed into a sample aliquot of water and were determined by flow injection analysis using a mass spectrometer. The fiber coatings and the developed electrochemical SPME method were found to be stable and reproducible (RSD < 5%; N = 5) and could be extended to several cations and anions, confirming the versatility of the approach. Preconcentration of the analyte on the fiber was also possible by repeating the processes to increase the amount of analyte extracted.

Letter↗
PubMed2003

Improved electrodeposited iridium oxide pH sensor fabricated on etched titanium substrates.

In the present paper, the preparation and characterization of an improved solid-state pH sensor are described. The sensor is based on anodically electrodeposited iridium oxide film, as a pH-sensing layer. Merits of the present sensor include (i) excellent adhesion of the pH sensitive layer to the substrate, (ii) excellent reproducibility of sensor fabrication, (iii) faster preparation procedure, and (iv) low cost of the titanium substrate. These advantages are realized by combining acid-etched titanium as the electrode substrate with an optimized electrodeposition solution consisting of IrCl4 as an iridium source, hydrogen peroxide, potassium oxalate, and potassium carbonate. Heating the electrodeposition solution to 90 degrees C reduced the time required for solution development from approximately 3 days to 10 min. The pH-sensing layer is protected with a layer of Nafion and a microporous polyester membrane. The improved sensor showed a super-Nernstian response (-73.7 +/- 1.2 mV/pH unit) in the pH range of 1.5-11.5. The present pH sensor, fabricated in a tubular form, is used as a detector in a flow injection analysis (FIA) system for pH measurements. Optimization of the FIA experimental parameters resulted in a linear dependence of peak heights on the pH of the injected samples in the pH range of 2-11.

Journal Article↗
PubMed2003

Advantages of soft versus hard constraints in self-modeling curve resolution problems. Alternating least squares with penalty functions.

A new algorithm for self-modeling curve resolution (SMCR) that yields improved results by incorporating soft constraints is described. The method uses least squares penalty functions to implement constraints in an alternating least squares algorithm, including nonnegativity, unimodality, equality, and closure constraints. By using least squares penalty functions, soft constraints are formulated rather than hard constraints. Significant benefits are (obtained using soft constraints, especially in the form of fewer distortions due to noise in resolved profiles. Soft equality constraints can also be used to introduce incomplete or partial reference information into SMCR solutions. Four different examples demonstrating application of the new method are presented, including resolution of overlapped HPLC-DAD peaks, flow injection analysis data, and batch reaction data measured by UV/visible and near-infrared spectroscopy (NIR). Each example was selected to show one aspect of the significant advantages of soft constraints over traditionally used hard constraints. Incomplete or partial reference information into self-modeling curve resolution models is described. The method offers a substantial improvement in the ability to resolve time-dependent concentration profiles from mixture spectra recorded as a function of time.

Journal Article↗
PubMed2004

Prussian blue based nanoelectrode arrays for H(2)O(2) detection.

We propose to form nanoelectrode arrays by deposition of the electrocatalyst through lyotropic liquid crystalline templates onto inert electrode support. Whereas Prussian Blue is known to be a superior electrocatalyst in hydrogen peroxide reduction, carbon materials used as electrode support demonstrate only a minor activity. We report on the possibility for nanostructuring of Prussian Blue by its electrochemical deposition through lyotropic liquid crystalline templates, which is noticed from atomic force microscopy images of the resulting surfaces. The resulting Prussian Blue based nanoelectrode arrays in flow injection analysis mode demonstrate a sub-part-per-billion detection limit (1 x 10(-)(8) M) and a linear calibration range starting exactly from the detection limit and extending over 6 orders of magnitude of H(2)O(2) concentrations (1 x 10(-)(8) to 1 x 10(-)(2) M), which are the most advantageous analytical performances in hydrogen peroxide electroanalysis.

Journal Article↗
PubMed2004

Electropolymerized flavin adenine dinucleotide as an advanced NADH transducer.

Electropolymerizing the prosthetic group (flavin adenine dinucleotide, FAD) responsible in the active sites of dehydrogenases for NAD(+)|NADH regeneration, we succeeded in mimicking enzyme activity. Poly(FAD) characterized by an additional polymer-type redox reaction has been discovered as a highly effective electrocatalyst for NADH oxidation: operating at the lowest potentials reported for NADH transducers (0.00 V, pH 7.4), poly(FAD) is characterized by the electrochemical rate constant of 1.8 +/- 0.6 x 10(-3) cm s(-1), which is at the level of the NADH mass-transfer constant. Flow injection analysis of NADH with the poly(FAD)-modified wall-jet electrode as a detector has been characterized by a linear calibration range prolonged down to 5 x 10(-7) M and a sensitivity of 0.08 A M(-1) cm(-2), which taking into account the dispersion coefficient ( approximately 3), is at the diffusion-limiting value. In contrast to the low molecular weight mediators able to exhibit similar electrocatalytic properties, poly(FAD)-modified electrodes are characterized by the dramatically improved stability and, thus, can be considered as the most advantageous NADH transducers for analytical chemistry.

Binding Sites↗
PubMed2006

Electrochemical detection of arsenic(III) using iridium-implanted boron-doped diamond electrodes.

Iridium-modified, boron-doped diamond electrodes fabricated by an ion implantation method have been developed for electrochemical detection of arsenite (As(III)). Ir+ ions were implanted with an energy of 800 keV and a dose of 10(15) ion cm(-2). An annealing treatment at 850 degrees C for 45 min in H2 plasma (80 Torr) was required to rearrange metastable diamond produced by an implantation process. Characterization was investigated by SEM, AFM, Raman, and X-ray photoelectron spectroscopy. Cyclic voltammetry and flow injection analysis with amperometric detection were used to study the electrochemical reaction. The electrodes exhibited high catalytic activity toward As(III) oxidation with the detection limit (S/N = 3), sensitivity, and linearity of 20 nM (1.5 ppb), 93 nA microM(-1) cm(-2), and 0.999, respectively. The precision for 10 replicate determinations of 50 microM As(III) was 4.56% relative standard deviation. The advantageous properties of the electrodes were its inherent stability with a very low background current. The electrode was applicable for analysis of spiked arsenic in tap water containing a significant amount of various ion elements. The results indicate that the metal-implanted method could be promising for controlling the electrochemical properties of diamond electrodes.

Arsenites↗
PubMed2006

Electrochemical oxidation of oxalic acid at highly boron-doped diamond electrodes.

Electrochemical oxidation of oxalic acid has been investigated at bare, highly boron-doped diamond electrodes. Cyclic voltammetry and flow injection analysis with amperometric detection were used to study the electrochemical reaction. Hydrogen-terminated diamonds exhibited well-defined peaks of oxalic acid oxidation in a wide pH range. A good linear response was observed for a concentration range from 50 nM to 10 microM, with an estimated detection limit of approximately 0.5 nM (S/N = 3). In contrast, oxygen-terminated diamonds showed no response for oxalic acid oxidation inside the potential window, indicating that surface termination contributed highly to the control of the oxidation reaction. An investigation with glassy carbon electrodes was conducted to confirm the surface termination effect on oxalic acid oxidation. Although a hydrogen-terminated glassy carbon electrode showed an enhancement of signal-to-background ratio in comparison with untreated glassy carbon, less stability of the current responses was observed than that at hydrogen-terminated diamond.

Boron↗
PubMed1996

Miniaturized biosensors employing electropolymerized permselective films and their use for creatine assays in human serum.

Miniaturized, disposable amperometric biosensors for determination of creatinine in human serum are described. The base electrodes are fabricated using micro-electronics techniques, to build a multilayer film structure on a polyimide foil. By using a thin electropolymerized film of poly(1,3-diaminobenzene), the electrochemical interferences from ascorbate, urate, acetaminophen, and other oxidizable species are greatly diminished. The multienzyme system (creatininase, creatinase, sarcosine oxidase) is immobilized on top of the permselective layer using cross-linking of the proteins with glutaraldehyde. The electropolymerization conditions for obtaining almost ideal permselectivity of the inner layer are defined, as well as the optimal enzyme layer preparation. A composite polymeric outer membrane [Nafion + poly-(2-hydroxy-ethyl methacrylate) is used for diffusion control and to protect the enzyme layer from fouling. The reagentless planar sensors for creatinine and creatine have fast response time (t95 = 1 min), linear response up to 1.2 mM in batch-type and 2.0 mM in flow injection analysis and a detection limit of 10-20 muM. They are applied in a differential setup for creatinine assay in control and hospital human serum samples and are suitable for incorporation in a portable analyzer.

Biosensing Techniques↗
PubMed1999

Sol-gel thin-film immobilized soybean peroxidase biosensor for the amperometric determination of hydrogen peroxide in acid medium.

An acid-stable soybean-peroxidase biosensor was developed by immobilizing the enzyme in a sol-gel thin film. Methylene blue was used as a mediator because of its high electron-transfer efficiency. The sol-gel thin film and enzyme membrane were characterized by FT-IR, and the effects of pH, operating potential, and temperature were explored for optimum analytical performance by using the amperometric method. The H2O2 sensor exhibited a fast response (5 s), high sensitivity (27.5 microA/mM), as well as good thermostability and long-term stability. In addition, the performance of the biosensor was investigated using flow-injection analysis (FIA).

Enzymes, Immobilized↗
PubMed1999

An electrochemiluminescence flow-through cell and its applications to sensitive immunoassay using N-(aminobutyl)-N-ethylisoluminol.

An electrochemiluminescence (ECL) flow-through cell with a carbon fiber electrode for ECL detection was developed, and a flow injection analysis system containing the cell was established. N-(aminobutyl)-N-ethylisoluminol (ABEI), an ECL reagent, was determined by this system. A straight-line calibration curve for ABEI (r = 0.999) was obtained from 6 fmol to 25 pmol. The detection limit of ABEI was 6 fmol (S/N = 2) and the relative standard deviation was 1.7% at 1.5 pmol (n = 10). The system was used for immunoassay of human immunogloblin G (hIgG) with ABEI-labeled anti-hIgG. A calibration curve of hIgG was obtained from 80 pg/mL to 1.3 ng/mL. The detection limit of hIgG was 80 pg/mL (S/N = 2), and the relative standard deviation was 1.8% at 0.5 ng/mL (n = 10). Sensitivity and accuracy for sera samples were found to considerably exceed those of the conventional methods, such as single-radial immunodiffusion and nepherometric immunoassay. The present system should prove useful for immunoassay.

Electrochemistry↗
PubMed1999

Improving data acquisition for fast-scan cyclic voltammetry.

Described is an improved data acquisition system for fast-scan cyclic voltammetry (FSCV). The system was designed to significantly diminish noise sources that were identified in previously recorded FSCV measurements for the detection of neurotransmitters. Minimized noise is necessary to observe the low concentrations of neurotransmitters that are physiologically important. The system was based on a high-speed, 16-bit AD/DA acquisition board that allowed high scan rates and better resolved the small faradaic currents which remained after background subtraction. Irregularities that occur when independent timing sources are used for generation of the voltage waveform and collection of the current can create large noise artifacts near the voltage limits during FSCV. These were eliminated by the use of a single acquisition board that generated the voltage waveform and collected the current. Noise from frequency drift of the power line was eliminated through the use of a phase-locked loop. To demonstrate the improved performance of the system, data were collected using carbon-fiber microelectrodes in a flow injection analysis system and in brain slices. This new data acquisition system performed significantly better than another system previously used in our laboratory without these features. The improved detection limits of the new system allowed clearly resolved current spikes featuring pre-release "feet" to be recorded adjacent to individual mast cells following chemical stimulation. When combined with false-color plots, the low-noise system facilitated identification of dopamine release in a freely moving animal.

Animals↗
PubMed2000

Electrochemical oxidation of histamine and serotonin at highly boron-doped diamond electrodes.

The electrochemistry of histamine and serotonin in neutral aqueous media (pH 7.2) was investigated using polycrystalline, boron-doped diamond thin-film electrodes. Cyclic voltammetry, hydrodynamic voltammetry, and flow injection analysis (FIA) with amperometric detection were used to study the oxidation reactions. Comparison experiments were carried out using polished glassy carbon (GC) electrodes. At diamond electrodes, highly reproducible and well-defined cyclic voltammograms were obtained for histamine with a peak potential at 1.40 V vs SCE. The voltammetric signal-to-background ratios obtained at diamond were 1 order of magnitude higher than those obtained for GC electrodes at and above 100 microM analyte concentrations. A linear dynamic range of 3-4 orders of magnitude and a detection limit of 1 microM were observed in the voltammetric measurements. Well-defined sweep rate-dependent voltammograms were also obtained for 5-hydroxytryptamine (5-HT). The characteristics of the voltammogram indicated lack of adsorption of its oxidation products on the surface. No fouling or deactivation of the electrode was observed within the experimental time of several hours. A detection limit of 0.5 microM (signal-to-noise ratio 13.8) for histamine was obtained by use of the FIA technique with a diamond electrode. A remarkably low detection limit (10 nM) was obtained for 5-HT on diamond by the same method. Diamond electrodes exhibited a linear dynamic range from 10 nM to 100 microM for 5-HT determination and a range of 0.5-100 microM for histamine determination. The FIA response was very reproducible from film to film, and the response variability was below 7% at the actual detection limits.

Boron↗