Enzyme-less amperometric biosensor for l-ascorbate using poly-l-histidine-copper complex as an alternative biocatalyst

A poly-l-histidine(PLH)-copper(II) complex can be used as an alternative biocatalyst in an O(2) detection-type amperometric enzyme-less l-ascorbate (AsA) sensor. The PLH-Cu(II) membrane was simply prepared by entrapping the PLH in polyacrylamide gel and subsequent treatment of the gel with CuCl(2) solution. This enzyme-less biosensor can be used over a relatively wide pH region from 4 to 11 and enables precise determination of AsA (RSD less than 3%, n=10) at pH 7.0. The fundamental performance characteristics (sensitivity, response time, and linear range) of this PLH-Cu(II)-based sensor is comparable to those of a native ascorbate oxidase-based sensor. Unfortunately, the selectivity is inherently rather low and, as a result, the response was degraded in the presence of higher concentrations (more than mM order) of quinones. However, reducing sugars caused no interference and the sensor could be used to detect AsA in some fruits and drinks. This enzyme-less sensor has excellent stability for at least 3 months of repeated analysis (more than 300 samples) without loss of ordinal activity.     

Ion-exclusion chromatography with conductimetric detection of aliphatic carboxylic acids on a weakly acidic cation-exchange resin by elution with benzoic acid-beta-cyclodextrin

In this study, an aqueous solution consisting of benzoic acid with low background conductivity and beta-cyclodextrin (beta-CD) of hydrophilic nature and the inclusion effect to benzoic acid were used as eluent for the ion-exclusion chromatographic separation of aliphatic carboxylic acids with different pKa values and hydrophobicity on a polymethacrylate-based weakly acidic cation-exchange resin in the H+ form. With increasing concentration of beta-cyclodextrin in the eluent, the retention times of the carboxylic acids decreased due to the increased hydrophilicity of the polymethacrylate-based cation-exchange resin surface from the adsorption of OH groups of beta-cyclodextrin. Moreover, the eluent background conductivity decreased with increasing concentration of beta-cyclodextrin in 1 mM benzoic acid, which could result in higher sensitivity for conductimetric detection. The ion-exclusion chromatographic separation of carboxylic acids with high resolution and sensitivity was accomplished successfully by elution with a 1 mM benzoic acid-10 mM cyclodextrin solution without chemical suppression.     

Potentiometric stripping determination of heavy metals using a graphite-reinforcement carbon vibrating electrode

Potentiometric stripping determination of Cd, Cu and Zn using a vibrating electrode (VE) is presented. A simple VE was constructed by using a piezoelectric bimorph oscillator and an inexpensive graphite-reinforcement carbon (GRC) rod (a mechanical pencil). Experimental results obtained with the simple VE follow the equation valid for quantitative application of potentiometric stripping analysis (PSA) in large solutions. It was found that the GRC vibrating electrode is suitable for multielement trace analysis of small samples of 0.05 mL. The relative standard deviations for divalent metals are better than 2%. Received: 3 February 1997 / Revised: 2 June 1997 / Accepted: 7 June 1997     

Determination of alkylphenols and alkylphenol polyethoxylates by reversed-phase high-performance liquid chromatography and solid-phase extraction

A simple, accurate and reproducible reversed-phase high-performance liquid chromatography (HPLC) method was developed for the separation and characterisation of alkylphenols (APs) and alkylphenol polyethoxylates (APEOs), using a C18 octadecyl silica (ODS) column. APs and each APEO oligomer were separated successfully within a reasonable time without gradient elution. An excellent resolution was obtained, even for mixtures of APs and low EO number APEOs, which are otherwise difficult to separate using conventional normal-phase HPLC methods. This method, combined with solid-phase extraction, was highly applicable for the simultaneous determination of alkylphenols and alkylphenol ethoxylates in real samples.     

Monolithic octadecylsilyl-silica gel column for the high-speed ion chromatographic determination of acidity

A monolithic ODS-silica gel column modified by saturating it with lithium dodecylsulfate (Li-DS) was used to demonstrate the high-speed separation of H+ from other mono- and divalent cations, such as Na+, NH4+, K+, Mg2+ and Ca2+ using ion chromatography (IC). Using a 5 mM EDTA-2K solution containing 0.10 mM Li-DS (pH 4.80) as eluent, H+ was eluted with a sharp and symmetrical peak within 1.0 min before other cations at a flow-rate of 1.5 ml min(-1). The rapid elution of H+ and its conductimetric detection could be attributed to the presence of EDTA (HY2-), which can convert H+ ions as anions. i.e. H(+) + H2Y(2-) --> H3Y(-). The acidity of rainwater and deionized water samples was determined using this IC system with satisfactory results.     

Determination of l-lactate by differential pulse polarography with covalently immobilized enzyme on Chitopearl

Summary The covalent bonding of l-lactate oxidase (no E.C.) to cross-linked copolymers of chitosan (Chitopearl) using the glutaraldehyde method is described and applied to the determination of l-lactate in serums of diseased and normal animals. The bioreactor packing of the immobilized enzyme is stable for at least 4 months and can be continuously used for the rapid and simple determination of l-lactate. The reduction current of pyruvate, corresponding to the product of enzymatic reaction, is determined by differential pulse polarography. The reproducibility (RSD) for 10 mol/l l-lactate is 0.78% (n=6) and the detection limit is 0.62 mol/l (k=2, confidence level 97.72%).     

Simultaneous ion-exclusion/cation-exchange chromatography of anions and cations in acid rain waters on a weakly acidic cation-exchange resin by elution with sulfosalicylic acid

A simple, selective, and sensitive method for the simultaneous determination of anions (sulfate, nitrate, and chloride) and cations (sodium, ammonium, potassium, magnesium, and calcium) in acid rain waters was developed using ion-exclusion/ cation-exchange chromatography with conductimetric detection. A weakly acidic cation-exchange resin column (Tosho TSKgel OA-PAK-A) and a sulfosalicylic acid-methanol-water eluent was used. With a mobile phase comprising 1.25 mM sulfosalicylic acid in methanol-water (7.5:92.5) at 1.2 ml/min, simultaneous separation and detection of the above anions and cations was achieved in about 30 min. Linear calibration plots of peak area versus concentration were obtained over the concentration ranges 0-1.0 mM for anions (R=0.9991) and 0-0.5 mM for cations (R=0.9994). Detection limits calculated at S/N=3 ranged from 4.2 to 14.8 ppb for the anions and from 2.4 to 12.1 ppb for the cations. The reproducibility of retention times was 0.14-0.15% relative standard deviation (RSD) for anions and 0.18-0.31% for cations, and reproducibility of chromatographic peak areas was 1.22-1.75% RSD for anions and 1.81-2.10% for cations. The method was applied successfully to the simultaneous determination of anions and cations in aerosols transported from mainland China to central Japan, as determined by a meteorological satellite data analyzer.     

Hemin-adsorbed carbon felt for sensitive and rapid flow-amperometric detection of dissolved oxygen

Hemin was physically adsorbed onto porous carbon felt (CF), a microelectrode ensemble of micro-carbon fiber (ca. 7 μm in diameter) and possessing a three-dimensional random structure. The hemin-CF exhibited a well-defined redox wave that is due to Fe(III)/Fe(II) redox process in hemin, with a formal potential of ?0.32 V (vs. Ag/AgCl) in deoxygenated buffer solution of pH 7.0. The surface coverage of the electroactive hemin molecules on the surface of the CF was calculated to be 5.0?×?10^?11 mol cm^?2, and the apparent heterogeneous electron transfer rate constant is 3.35 s^?1. The hemin-CF electrode displays excellent electrocatalytic activity for the reduction of dissolved oxygen (DO), and the magnitude of the cathodic current increases with increasing concentrations of DO in the sample solution. The electrode was used as a flow-through detector for sensitive and rapid consecutive determination of DO. Deoxygenated pH 7.0 solutions were analyzed at a flow rate of 8.0 mL min^?1 at an applied potential of ?0.2 V, and highly reproducible cathodic peak current responses to DO were observed in the 0.72 to 13.3 mg L^?1 concentration range. The maximum throughput is 170 samples h^?1. The hemin-CF-based amperometric flow-sensor was applied to determine the concentration of DO in environmental water samples.

Development of Plasma Window for gas charge stripper at RIKEN RIBF

A differential pumping system with a Plasma Window (PW) has been developed for an application to a window-less He gas charge stripper. A PW with Southern Methodist University design was newly fabricated and tested off-line for evaluation of differential pumping efficiency. Switching gases for plasma seed from Ar to He was tested. It was found that the PW reduced the pressure at the first differential pumping section to 1/13 for Ar and to 1/17 for He, comparing with differential pumping without PW. At the second pumping section, the pressures were drastically reduced to 1/160 and to 1/4,520 for Ar and He, respectively. These pressure reduction factors indicate that one differentially pumped stage could be removed from a conventional differential pumping system.     

Molecular Insights into the Ligand-Based Six-Proton- and Six-Electron-Transfer Processes Between Tris-ortho-Phenylenediamines and Tris-ortho-Benzoquinodiimines

The global demand for energy and the concerns over climate issues renders the development of alternative renewable energy sources such as hydrogen (H₂) important. A high-spin (hs) Fe^II complex with o-phenylenediamine (opda) ligands, [Fe^II(opda)₃]²⁺ (hs- [6R] ²⁺), was reported showing photochemical H₂ evolution. In addition, a low-spin (ls) [Fe^II(bqdi)₃]²⁺ (bqdi: o-benzoquinodiimine) (ls- [0R] ²⁺) formation by O₂ oxidation of hs- [6R] ²⁺, accompanied by ligand-based six-proton and six-electron transfer, revealed the potential of the complex with redox-active ligands as a novel multiple-proton and -electron storage material, albeit that the mechanism has not yet been understood. This paper reports that the oxidized ls- [0R] [PF₆]₂ can be reduced by hydrazine giving ls-[Fe^II(opda)(bqdi)₂][PF₆]₂ (ls- [2R] [PF₆]₂) and ls-[Fe^II(opda)₂(bqdi)][PF₆]₂ (ls- [4R] [PF₆]₂) with localized ligand-based proton-coupled mixed-valence (LPMV) states. The first isolation and characterization of the key intermediates with LPMV states offer unprecedented molecular insights into the design of photoresponsive molecule-based hydrogen-storage materials.