AFM studies of DNA structures on mica in the presence of alkaline earth metal ions

As counterions of DNA on mica, Mg2+, Ca2+, Sr2+ and Ba2+ were used for,clarifying whether DNA molecules equilibrate or are trapped on mica surface. End to end distance and contour lengths were determined from statistical analysis of AFM data. It was revealed that DNA molecules can equilibrate on mica when Mg2+, Ca2+ and Sr2+ are counterions. When Ba2+ is present, significantly crossovered DNA molecules indicate that it is most difficult for DNA to equilibrate on mica and the trapping degree is different under different preparation conditions. In the presence of ethanol, using AFM we have also observed the dependence of B A conformational transition on counterion identities. The four alkaline earth metal ions cause the B-A transition in different degrees, in which Sr2+ induces the greatest structural transition.

Fabrication of metalloporphyrin-polyoxometalyte hybrid film by layer-by-layer method and its catalysis for dioxygen reduction

Through layer-by-layer method [tetrakis(N-methylpyridyl)porphyrinato] cobalt (CoTMPyP) and polyoxometalyte were alternately deposited on 4-aminobenzoic acid (4-ABA) modified glassy carbon electrode. The resulting organic-inorganic hybrid films were characterized by cyclic voltammetry (CV), UV/visible absorption spectroscopy, and atomic force microscopy (AFM). It was proved that the multilayer films are uniform and stable. CoTMPyP-containing multilayer films exhibit remarkable electrocatalytic activity for the reduction of O-2. Rotating disk electrode (RDE) voltammetry and rotating ring-disk electrode (RRDE) voltammetry confirm that P2W18/CoTMPyP multilayer films can catalyze the four-electron almost reduction of O-2 to water in pH > 4.0 buffer solution, while SiW12/CoTMPyP multilayer films catalyze about two-electron reduction of O-2 to H2O2 in pH 1 - 6 buffer solutions. The kinetic constants for O-2 reduction were comparatively investigated at P2W18/CoTMPyP and SiW12/CoTMPyP multilayer films electrodes.

Capillary electrophoresis with solid-state electrochemiluminescence detector

We report capillary electrophoresis coupling to a solid-state electrochemiluminescence (ECL) detector for the first time. The solid-state ECL detector was fabricated by immobilizing the ECL reagent tris(2,2'-bipyridyf)ruthenium (TBR) in poly-(p-styrenesulfonate)-silica-poly(vinyl alcohol) grafting 4-vinylpyridine copolymer films. The excellent stability of the solid-state ECL detector in the phosphate solution satisfied application in CE. The CE with solid-state ECL detector system was characterized using tripropylamine (TPA) and proline. The influences of detection potential, the concentration of TBR in the film, and pH value of ECL buffer were investigated. The linear range for TPA and proline was 0.005-10 muM and 5-10 mM with correlation coefficients of 0.997 and 0.998, respectively. The detection limit (signal-to-noise ratio S/N = 3) was estimated to be 0.002 and 2.0 muM for TPA and proline, respectively. The relative standard deviations for 1.0 pm TPA and 1.0 mm proline were 8.7% and 7.5% with theoretical plate numbers of 70 000 and 16 000, respectively. Compared with the CE-ECL of TBR in aqueous solution, the CE coupling with solid-state ECL detector system gave the same sensitivity of analysis.

Capillary electrophoresis with amperometric detection of curcumin in Chinese herbal medicine pretreated by solid-phase extraction

In the present study, curcumin from Chinese herbal medicine turmeric was determined by capillary electrophoresis with amperometric detection (CE-AD) pretreated by a self-designed, simple, inexpensive solid-phase extraction (SPE) cartridge based on the material of tributyl phosphate resin. An average concentration factor of 9 with the recovery of >80% was achieved when applied to the analysis of curcumin in extracts of turmeric. Under the optimized CE-AD conditions: a running buffer composed of 15 mM phosphate buffer at a pH 9.7, separation voltage at 16 W, injection for 6 s at 9 W and detection at 1.20 V, CE-AD with SPE exhibited low detection limit as 3 - 10(-8) mol/l (SIN = 3), high efficiency of 1.0(.)10(5) N, linear range of 7(.)10(-4) -3(.)10(-6) mol/l (r = 0.9986) for curcumin extracted from light petroleum. The method developed resulted in enhancement of the detection sensitivity and reduction of interference from sample matrix in complicated samples and exhibited the potential application for routine analysis, especially in food, because a relatively complete process of sample treatment and analysis was described.

Determination of sulpiride by capillary electrophoresis with end-column electrogenerated chemiluminescence detection

Background: Capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)(3)(2+)]-electro-generated chemiluminescence (ECL) detection is a promising method for clinical analysis. In this study, a method combining CE with Ru(bpy)(3)(2+) ECL (CE-ECL) detection that can be applied to amine-containing clinical species was developed, and the performance of CE-ECL as a quantitative method for determination of sulpiride in human plasma or urine was evaluated. Methods: Sulpiride was separated by capillary zone electrophoresis in uncoated fused-silica capillaries [510 cm x 25 mum (i.d.)] filled with phosphate buffer (pH 8.0 and a driving voltage of +15 kV, with end-column Ru(bpy)(3)(2+) ECL detection. A platinum disc electrode was used as working electrode. Sulpiride in human plasma or urine samples (100 muL) was extracted by a double-step liquid-liquid extraction procedure, dried under nitrogen at 35 degreesC in a water bath, and reconstituted with 100 muL of filtered water. The extraction solvent was ethyl acetate-dichloromethane (5:1 by volume). Results: Under optimum conditions (pH 8.0 phosphate buffer, injection for 6 s at 10 kV, and +1.2 V as detection potential), separation of sulpiride was accomplished within 4 min. The calibration curve was linear over a concentration range of 0.05-25.0 mumol/L, and the limit of detection was 2.9 x 10(-8) mol/L for sulpiride. Intra- and interday CVs for ECL intensities were <6%. Extraction recoveries of sulpiride were 95.6-101% with CVs of 2.9-6.0%. The method was,clinically validated for patient plasma and urine samples. Conclusions: CE combined with Ru(bpy)(3)(2+) ECL is reproducible, precise, selective, and enables the analysis of sulpiride in human plasma and urine. It thus is of value for rapid and efficient analysis of amine-containing analytes of clinical interest.

Amplified immunoassay of human IgG using real-time biomolecular interaction analysis (BIA) technology

An automated biomolecular interaction analysis instrument (BI-Acore) based on surface plasmon resonance (SPR) has been used to determine human immunoglobulin G (IgG) in real time. Polyclonal anti-human IgG antibody was covalently immobilized to a carboxymethyldextran modified gold film surface. The samples of human IgG prepared in HBS buffer were poured over the immobilized surface. The signal amplification antibody was applied to amplify the response signal. After each measurement, the surface was regenerated with 0.1 mol/L H3PO4. The assay was rapid, requiring only 30 min for antibody immobilization and 20 min for each subsequent process of immune binding, antibody amplification and regeneration. The antibody immobilized surface had good response to human IgG in the range of 0.12-60 nmol/L with a detection limit of 60 pmol/L. The same antibody immobilized surface could be used for more than 110 cycles of binding, amplification and regeneration. The results demonstrate that the sensitivity, specificity and reproducibility of amplified immunoassay using real-time BIA technology are satisfactory.

Synthesis of tetra-iron substituted sandwich-type pentadecatungstodiarsenate and its multilayers on glassy carbon electrode by self-assembly with electrocatalytic properties

A novel sandwich-type compound, Na-12 [Fe-4 (H2O)(2) (As2W15O56)(2)] . 38H(2)O (denoted as Fe(4)AS(4)W(30)) was synthesized. The compound was well characterized by means of IR, UV-Vis, W-138 NMR and elemental analyses. Redox electrochemistry of the compound has been studied in acid buffer solutions using cyclic voltammetry(CV). The compound containing multilayer films has been fabricated on the 4-aminobenzoic acid(4-ABA) modified glassy carbon electrode(GCE) surface by alternate deposition with a quaternized poly (4-vinylpyridine) partially completed with [Os(bpy)(2)Cl](2+/-) (denoted as QPVP-Os). CV, X-ray photoelectron spectroscopy (XPS) and UV-Vis spectroscopy were used to characterize the asprepared multilayer films. It is proved that the multilayer films are uniform and stable. The electrocatalytic activities of the multilayer films were investigated on the reduction of two substrates of important analytical interest, NO2- and H2O2.

Enhanced surface plasmon resonance immunoassay for human complement factor 4

Using an enhanced surface plasmon resonance (SPR) immunosensor, we have determined the concentration of human complement factor 4 (C4). Antibody protein was concentrated into a carboxymethyldextran-modified gold surface by electrostatic attraction force and a simultaneous covalent immobilization of antibody based on amine coupling reaction took place. The sandwich method was applied to enhance the response signal and the specificity of antigen binding assay. The antibody immobilized surface had good response to C4 in the range of 0.02-20 mug/ml by this enhanced immunoassay. The regeneration effect by pH 2 glycine-HC1 buffer was also investigated. The same antibody immobilized surface could be used more than 80 cycles of C4 binding and regeneration. In addition, the ability to determinate C4 directly from serum sample without any purification was investigated. The sensitivity, specificity and reproducibility of the enhanced immunoassay are satisfactory. The results clearly demonstrate the advantages of the enhanced SPR technique for C4 immunoassay.

Bienzymatic amperometric biosensor for glucose based on polypyrrole/ceramic carbon as electrode material

A novel amperometric biosensor utilizing two enzymes, glucose oxidase (GOD) and horseradish peroxidase (HRP), was developed for the cathodic detection of glucose. The glucose biosensor was constructed by electrochemical formation of a polypyrrole (PPy) membrane in the presence of GOD on the surface of a HRP-modified sol-gel derived-mediated ceramic carbon electrode. Ferrocenecarboxylic acid (FCA) was used as mediator to transfer electron between enzyme and electrode. In the hetero-bilayer configuration of electrode, all enzymes were well immobilized in electrode matrices and showed favorable enzymatic activities. The amperometric detection of glucose was carried out at +0.16 V (versus saturated calomel reference electrode (SCE)) in 0.1 M phosphate buffer solution (pH 6.9) with a linear response range between 8.0 x 10(-5) and 1.3 x 10(-3) M glucose. The biosensor showed a good suppression of interference in the amperometric detection.

Single-crystal X-ray structure and properties of K-10[Ni-4(H2O)(2)(AsW9O34)(2)]center dot 4H(2)O

The rational synthesis and the structural and magnetic characterization of a nickel cluster are presented. The compound comprises a rhomblike Ni4O16 group encapsulated between two-heptadentate tungstoarsenate ligands [AsW9O34](9-). The crystal structure of K-10[Ni-4(H2O)(2)(AsW9O34)(2)](.)4H(2)O was solved in monoclinic, P2(1)/n symmetry, with a = 12.258(3) Angstrom, b = 21.232(4) Angstrom, c = 15.837(3) Angstrom, beta = 92.05(3)degrees, V = 4119.1(14) Angstrom(3), Z = 2, and R = 0.0862. The crystal structure of the Ni(II) derivative was compared with that of the Cu(II), Zn(II), Co(II) and Mn(II) derivatives. The Ni4O14(H2O)(2) unit in the compound shows no Jahn-Teller distortion. On the other hand, the Ni(II) derivative shows ferromagnetic exchange interactions within the Ni4O16 group (J = 7.8 cm(-1), J' = 13.7 cm(-1)) and an S = 4 ground state, the highest spin state reported in a heteropoly complex. Its redox electrochemistry has been studied in acid buffer solutions using cyclic voltammetry. It exhibited two steps of one-electron redox waves attributed to redox processes of the tungsten-oxo framework. The new catalyst showed an electrocatalytic effect on the reduction of NO2-.

Determination of three beta-blockers by capillary electrophoresis with end-column electrochemical detection

Three beta -blockers (propranolol, timolol, acebutolol) were separated by capillary electrophoresis (CE) and detected by end-column electrochemical detection (EC). In the present work, a carbon fiber (33 mum) electrode was used as the working electrode. The effect of the buffer concentration, buffer pH, detection potential and separation voltage on the separation of analytes and behavior of electrochemical detection was systematically investigated. The optimum conditions determined were as following: 40 cm length, 25 mum i.d. capillary; 17.5 kV separation voltage; 2 s injection at 15 kV; 70mM phosphate buffer, pH 3.5; detection potential + 1.2V (vs. Ag/AgCl). Under these conditions, the linear ranges of beta -blockers were over three orders of magnititude and the low detection limit of 10(-8)M was obtained. This method was also applied to detect the simulated urine sample.

Electrocatalytic oxidation of NADH by rutin in biomembrane-like films on glassy carbon electrode

Stable lipid film was made by casting dipalmitoylphosphatidylcholine (DPPC) and rutin onto the surface of a glassy carbon (GC) electrode. The electrochemical behavior of rutin in the DPPC film was studied. The modified electrode coated with rutin gave quasi-reversible reduction-oxidation peak on cyclic voltammogram in the phosphate buffer (pH 7.4). The peak current did not decrease apparently after stored at 4 degreesC for 8 hours in refrigerator. This model of biological membrane was used to investigate the oxidation of dihydronicotinamide adenine dinucleotide (NADH) by rutin. Rutin in the film acts as a mediator. The modified electrode shows a great enhancement and the anodic peak potential was reduced by about 220 mV in the oxidation of 5 X 10(-3) mol L-1 NADN compared with that obtained at a bare glassy carbon electrode. (C) 2000 Elsevier Science S.A. All rights reserved.

Conformational transition of DNA in electroreduction studied by in situ UV and CD thin layer spectroelectrochemistry

Electrochemically induced three conformational transitions of calf thymus DNA from B-10.4 to Z(10.2)-DNA and from B-10.2 to B-10.4 and to C-DNA in 10 mM phosphate buffer solution (pH 7.21) at glassy carbon electrode are found and studied by in situ circular dichroism (CD) thin layer spectroelectrochemistry with singular value decomposition least square (SVDLS) analysis. It indicates that the so-called B-10.2 form and the C-form of DNA may be composed of B-10.4 and left-A DNA and of B-10.4 and right-A DNA, respectively. The irreversible electrochemical reduction of adenine and cytosine groups in the DNA molecule is studied by UV-Vis spectroelectrochemistry. Some electrochemical parameters alphan = 0.17, E-0' = -0.70 V (vs. Ag/AgCl), and the standard heterogeneous electron transfer rate constant, k(0) = 1.8 x 10(-5) cm s(-1) are obtained by double logarithmic analysis and non-linear regression. (C) 2000 Elsevier Science B.V. All rights reserved.

Anodic voltammetric behavior of medecamycin at a glassy carbon electrode

The anodic voltammetric behavior of medecamycin (MD) in the presence of various electrolytes was studied by linearsweep voltammetry, differential-pulse voltammetry and cyclic voltammetry at a glassy carbon electrode. In phosphate buffer solutions (pH = 9.4), MD is oxidized irreversibly. The peak potential is at about +0.75 V (vs.Ag/AgCl). The height of the peak is linearly increased with the concentration of MD over the range of 5 x 10(-5) similar to 1 x 10(-1) g/L. The method has been used for the direct determination of MD in tablets. The relative standard deviation (n = 10) is 1.8%. The recoveries of MD in urine samples are in the range of 95% similar to 115%.

Simultaneous determination of 2-aminothiazole, 2-aminobenzothiazole and 2-mercaptobenzothiazole by capillary electrophoresis with end-column amperometric detection

Capillary electrophoresis with amperometric detection is evaluated for the simultaneous determination of 2-aminothiazole (A), 2-amino-benzothiazole (AB), 2-mercaptobenzothiazole (AM). The cyclic voltammogram, hydrodynamic voltammogram, effect of pH, concentration of buffer and separation voltage on the separation and the detection were studied. The conditions were optimized as follows: 50 mM phosphate buffet; pH 6.0, 2s at 17.5 kV sample injection, separation at 17.5 kV, 1.2 V as detection potential. The method provided low detection limit as 0.5 mu M, 0.05 mu M and 0.01 mu M, wide linear range 2-200 mu M, 10-200 mu M and 0.025-100 mu M for A, AB, and AM, respectively. The variations in peak current and migration time for 15 consecutive injections of a standard containing 5 mu M each compound were 3.7, 2.1, and 3.9%, and 1.2, 0.8, and 1.2%, for A, AB and AM, respectively. This method was employed to analyze river water.