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41.
A rapid method of capillary zone electrophoresis is described to determine the oxidized (GSSG) and reduced (GSH) form of glutathione in plant tissue. In order to separate both analytes in a fused-silica capillary, the pH and composition of the electrolyte solution were optimized. The electrolyte composition was 100 mmol/L, borate 25 mmol/L Tris, and 0.2% w/v metaphosphoric acid (MPA), pH 8.2. Some instrumental conditions used to run the samples were hydrostatic injection for 30 s, 30 kV applied voltage, and UV detection (185 nm) at 25 degrees C. Linearity and useful range obtained for the calibration curves were optimum, with correlation coefficients about 0.999 in the 0-120 micromol/L range. The migration time was highly reproducible, less than 5 min being afforded to run a sample. Electrolyte buffer and samples required a careful pH control for optimal separation of both analytes. This aspect constitutes a critical analytical step when acids are used in the procedure for sample preparation. Simultaneous analysis of GSH and GSSG may provide a useful tool for comparative studies of plants in order to select those species with a potential capacity for detoxification from toxic elements or those appearing promising from phytoremediation for these elements. 相似文献
42.
A strategy for design of bioimprinted proteins with glutathione peroxidase (GPX) activity has been proposed. The proteins imprinted with a glutathione derivative were converted into selenium-containing proteins by chemical modifying the reactive hydroxyl groups of serines followed by sodium hydrogen selenide displacement. These selenium-containing proteins exhibited remarkable GPX activities and the GPX activities of reduction of H2O2 by glutathione (GSH) were found to be 101-817 U μmol−1, which approaches the activity of a selenium-containing catalytic antibody elicited by a hapten similar to our template. The steady state kinetic study for imprinted protein catalysis revealed Michaelis-Menten kinetics for both H2O2 and GSH, e.g. the pesudo-first-order rate constant kcat (H2O2) and the apparent Michaelis constant Km (H2O2) at 1 mM GSH were calculated to be 784 min−1 and 1.24×10−3 M, respectively, and the apparent second-order rate constant kcat (H2O2)/Km (H2O2) was determined to be 6.33×105 (M min)−1. The kinetics and the template inhibition showed that the strategy might be a remarkably efficient one for generating artificial enzyme with GPX activity. 相似文献
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《Analytical letters》2012,45(3):483-494
Abstract A new biosensor for the amperometric detection of hydrogen peroxide was developed by means of immobilized horseradish peroxidase (HRP) on a platinum disk based on gold nanoparticles, nafion, polythionine (PTn), and gelatin as matrixes. The mediator (PTn) was embedded in nafion film effectively without leaching even after long periods of operation, the immobilization of the enzyme comes from the cooperative binding by the Au nanoparticles and gelatin. The fabrication procedure of the biosensor was characterized by using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The electrochemical characteristics of the enzyme electrode with respect to the effect of pH, temperature, and the operational and storage stabilities were studied. The test demonstrated that the biosensors show high stability, fast response (<20 s), and a working range 0.05 to 30.6 mM (correlation coefficient: 0.9986), a detection limit of 0.02 mM to hydrogen peroxide (H2O2). The analytical results by this approach were in satisfactory agreement with those by conventional methods of titration. 相似文献
46.
《Analytical letters》2012,45(7-8):863-873
Abstract Total sulfur (S) concentration in biological samples was determined simultaneously with metal concentrations by inductively coupled plasma-atomic emission spectrometry (ICP). A 0.2 g portion of liver and other tissues were wet-digested with 1.0 ml mixed acid (HNO3 : HCLO4 = 5 : 1, v/v) at 130 – 150 °C. The solution was concentrated to about 0.1 ml and then diluted to 5.0 ml with double distilled water. Concentration of S was determined by ICP using ammonium sulfate as a standard S compound. Sulfur and other element concentrations in an NBS standard reference material (Bovine Liver SRM 1577) were within the certified values by this method. Concentrations of total S, cadmium (Cd), copper (Cu) and zinc (Zn) in the liver, kidney, spleen, lung, pancreas and blood serum were compared between the control and Cd-exposed rats. The three metal concentrations were increased significantly by Cd exposure. However, S concentration was not altered significantly in the liver and other tissues despite the extensive induction of metallothionein (MT) by the repeated Cd exposure. Metallothionein induced by the accumulated Cd (121 μg/g) and Zn (48 μg/g) in the liver was estimated to account for at maximum 7 % of the total S by assuming that the increased metals were all bound to MT. Concentration of S in blood serum was decreased significantly by Cd loading. 相似文献
47.
《Analytical letters》2012,45(10):1230-1241
In this study, anti-carbofuran monoclonal antibodies (Ab) were immobilized onto a gold electrode surface modified with multilayers of L-cysteine and gold colloidal nanoparticles (GNPs). Furthermore, horseradish peroxidase (HRP) as enzyme membrane was used for blocking unspecific sites and amplifying signal. The conformational properties of the immunosensor were characterized using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The concentration of antibody solution, pH of working buffer and incubation time were studied in detail for optimization of analytical performance. Under optimal conditions, the variation of current response was proportional to the concentration of carbofuran which ranged from 0.01 ng/mL to 50 ng/mL with a correlation coefficient of 0.9912. The detection limit was 0.01 ng/mL (S/N = 3). The proposed immunosensor exhibited good reproducibility and stability and it can be used for the rapid detection of carbofuran pesticide. 相似文献
48.
Subcellular distribution of Superoxide dismutase (SOD), catalase (CAT), selenium (SC) dependent glutathione peroxidase, and
Se-independent glutathione peroxidase (GSH-Px) activities were detected in different tissues (hepatopancreas, muscle, and
gill) of freshwater prawnsMacrobrachium malcolmsonii andMacrobrahium lamarrei lamarrei. CAT and SOD were found almost equally between the mitochondrial and cytosolic fraction. Both Se-dependent and Se-independent
GSH-Px activities were mainly found in cytosolic fraction. 相似文献
49.
Eliana M. Alhadeff Andrea M. Salgado Nei Pereira Jr. Belkis Valdman 《Applied biochemistry and biotechnology》2004,113(1-3):125-136
An automated flow injection analysis (FIA) system for quantifying ethanol was developed using alcohol oxidase, horseradish
peroxidase, 4-aminophenazone, and phenol. A colorimetric detection method was developed using two different methods of analysis,
with free and immobilized enzymes. The system with free enzymes permitted analysis of standard ethanol solution in a range
of 0.05–1.0 g of ethanol/L without external dilution, a sampling frequency of 15 analyses/h, and relative SD of 3.5%. A new
system was designed consisting of a microreactor with a 0.91-mL internal volume filled with alcohol oxidase immobilized on
glass beads and an addition of free peroxidase, adapted in an FIA line, for continued reuse. This integrated biosensor-FIA
system is being used for quality control of biofuels, gasohol, and hydrated ethanol. The FIA system integrated with the microreactor
showed a calibration curve in the range of 0.05–1.5 g of ethanol/L, and good results were obtained compared with the ethanol
content measured by high-performance liquid chromatography and gas chromatography standard methods. 相似文献
50.
Jing Juan XU Gang WANG Qing ZHANG Xing Hua XIA Hong Yuan CHEN? The Laboratory of Life Analytical Chemistry Department of Chemistry Nanjing University Nanjing 《中国化学快报》2005,16(4)
Carbon nanotube (CNT) is a novel carbon material discovered by Iijima1. It had beenfound to have the ability to promote electron transfer reactions when it was used tofabricate electrodes for the oxidation of biomolecules2-4. Just recently, the interest indemonstrating CNT for biosensing applications is now emerging5-10. However, most ofthese biosensors were based on CNT paste electrode or CNT modified glassy carbonelectrode by casting technology. Here, we reported another assembly… 相似文献