A flow injection system, comprising a biosensor based on a screen-printed carbon electrode containing Meldola’s Blue-Reinecke salt coated with glucose dehydrogenase, for the measurement of glucose

A biosensor for the measurement of glucose in serum has been developed, based on a screen-printed carbon electrode modified with Meldola’s Blue-Reinecke salt, coated with the enzyme glucose dehydrogenase (from Bacillus sp.), and nicotinamide adenine dinucleotide coenzyme (NAD⁺). A cellulose acetate layer was deposited on top of the device to act as a permselective membrane. The biosensor was incorporated into a commercially available, thin-layer, amperometric flow cell operated at a potential of only +0.05 V versus Ag/AgCl. The mobile phase consisted of 0.2 M phosphate buffer (pH 7.0) containing 0.1 M potassium chloride solution, and a flow rate of 0.8 ml min⁻¹ was used throughout the investigation. The biosensor response was linear over the range of 0.075-30 mM glucose, with the former representing the detection limit. The precision of the system was determined by carrying out 20 repeat injections of a 5-mM glucose standard, and the calculated coefficient of variation was 3.9%. It was demonstrated that this biosensor system could be applied to the direct measurement of glucose in serum without pretreatment. Therefore, this would allow high-throughput analysis, at low cost, for this clinically important analyte.     

I. Novel capacitative electrode with a wide frequency range for measurements of flash-induced changes of interface potential at the oil-water interface Mechanical construction and electrical characteristics of the electrode

The mechanical construction and the electrical properties of a new type of capacitative electrode for the oil-water interface are described. The electrode is designed to detect changes of the interface potential induced by photochemical, photophysical, and photobiological reactions occurring at the interface. The construction is based on capacitative coupling of two aqueous compartments separated by a thin Teflon film. Thereby, the oil-water interface is in horizontal position and the electrode is placed with its planar bottom about 10 μm above the interface. A main feature of the electrode is the transparency to visible light which is achieved by having a clear electrolyte solution in the inner compartment of the capacitative electrode.The aqueous subphase and the inner electrolyte are connected with AglAgCl electrodes to voltage amplifiers. The capacitative electrode is best operated under open circuit conditions. The frequency range experimentally verified is 500 $\text{MHz}\text{\$}\text{?}\text{?}{}_{\mathrm{<mtext>3</mtext><mtext>dB</mtext>}}\text{\$}\text{?}\text{0.03}\text{Hz}$. The sensitivity is mainly determined by the noise of the electronic amplifiers, typical 50–100 μV.     

Structure and I2/I redox catalytic behaviour of PEDOT-PSS films electropolymerized in aqueous medium: Implications for convenient counter electrodes in DSSC

The present paper focuses on the impact of different electro-synthesis processes, used to produce poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT-PSS) systems in aqueous medium, on the polymer backbone and its electro-catalytic activity.FE-SEM and micro-Raman spectroscopy were used to analyze the samples obtained by varying the conditions of the electro-polymerization processes. The I₂/I⁻ redox behaviour of various PEDOT-PSS layers has been comparatively examined in relation to the surface morphology and structural characteristics of the polymer. A response similar to that of a conventional Pt electrode has been revealed for the films produced by chronoamperometric technique.The study has demonstrated that the polymer structure plays an important role in the electro-activity, providing a possibility to improve the redox performance of PEDOT-PSS films by a proper choice of the synthesis methods. Moreover, the use of aqueous solution as a reaction medium can be welcomed as a sustainable chemistry way involving the use of environmentally friendly chemicals.     

人工湿地-微生物燃料电池耦合系统的研究进展

人工湿地-微生物燃料电池耦合系统(CW-MFC)是一种将人工湿地技术(CW)和微生物燃料电池技术(MFC)结合在一起的新型污水处理系统,其产电机理是产电微生物在底层湿地(阳极)的厌氧条件下生成电子,通过外电路传递到表面湿地(阴极)完成氧化还原反应。但是,近几年来,关于CW-MFC研究的文章较少且研究深度较浅。综述了电极材料、水力条件、湿地植物及微生物等条件对CW-MFC污水处理能力和产电能力的影响。在电极材料方面,选用导电性、吸附性及有效面积大的材料作为电极可有效提高CW-MFC产电与去污能力;在水利条件方面,在HRT为2-3 d的条件下,应选用升流式或升流-降流式的入水方式;湿地植物方面,种植湿地植物的CW-MFC在去污和产电能力上都要优于未种植植物的CW-MFC;微生物方面,阴极与阳极的微生物群落结构存在明显的差异,但存在的产电菌的种类却十分相似。CW-MFC中存在的常见产电微生物主要包括地杆菌属(Geobacter)、脱硫叶菌属(Desulfobulbus)、假单胞菌属(Pseudomona)和脱硫弧菌属(Desulfovibrio)等。最后对CW-MFC的研究方向进行了分析,以期为CW-MFC的实际应用提供理论依据。     

A novel biosensor based on l-homocysteine desulfhydrase enzyme immobilized in eggshell membrane

A novel biosensor for homocysteine determination has been developed. The biosensor was fabricated with l-homocysteine desulfhydrase immobilized on the ammonium selective electrode by means of eggshell membrane. The measurement principle is based on determination of ammonia due to the enzymatic reaction in the medium by ammonium selective electrode. The effects of enzyme loading, glutaraldehyde concentration, pH, buffer concentration, temperature, dithiotreitol (DTT) concentration and ionic strength adjustment buffer (ISA) on the biosensor response were investigated in detail. The linear detection range and limit of detection (LOD) for homocysteine were found to be 0.15–1.8 mM and 55 μM, respectively. Finally, the homocysteine biosensor has been applied to plasma samples for determination of total homocysteine contents.     

Comparison of methods for measuring oxygen consumption in tumor cells in vitro

The oxygen consumption rate of tumor cells affects tumor oxygenation and response to therapies. Highly sensitive methods for determining cellular oxygen consumption are, therefore, needed to identify treatments that can modulate this parameter. We compared the performances of three different methods for measuring cellular oxygen consumption: electron paramagnetic resonance (EPR) oximetry, the Clark electrode, and the MitoXpress fluorescent assay. To compare the assays, we used K562 cells in the presence of rotenone and hydrocortisone, compounds that are known to inhibit the mitochondrial electron transport chain to different extents. The EPR method was the only one that could identify both rotenone and hydrocortisone as inhibitors of tumor cell oxygen consumption. The Clark electrode and the fluorescence assay demonstrated a significant decrease in cellular oxygen consumption after administration of the most potent inhibitor (rotenone) but failed to show any significant effect of hydrocortisone. EPR oximetry is, therefore, the most sensitive method for identifying inhibitors of oxygen consumption on cell assays, whereas the Clark electrode offers the unique opportunity to add external compounds during experiments and still shows great sensitivity in studying enzyme and chemical reactions that consume oxygen (non-cell assays). Finally, the MitoXpress fluorescent assay has the advantage of a high-sample throughput and low bulk requirements but at the cost of a lower sensitivity.     

新型青霉素生物传感器的研究

以青霉素为模板分子,采用溶胶-凝胶法合成分子印迹膜,以浸泡的方法移除印迹分子,制备青霉素分子印迹膜电极。本印迹电极能有效地避免类似物对其测定的干扰。通过循环伏安法研究传感器对青霉素的响应特性,结果表明:富集时间为200 s,在0.1~1.8μg/L质量浓度范围内,青霉素在磷酸缓冲液(PBS)中的电流强度与其浓度呈良好的线性关系。吸附后的膜电极用甲醇洗脱后再生,可以重复利用3次,可以应用到实际检测中。     

Determination of folic acid by high‐performance liquid chromatography with direct electrogenerated chemiluminescence reaction

In our present work, it was found that the electrooxidation of folic acid (FA) was accompanied by electrogenerated chemiluminescence (ECL) emission. Out of the four inorganic salts, NaNO₃ solution was found to be a suitable supporting electrolyte for the ECL emission of FA. Coupled with high‐performance liquid chromatography separation, this simple ECL method was used for post‐column determination of FA. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of FA in the range of 1.0 × 10^?7 to 1.0 × 10^?5 g/mL and the detection limit was 5 × 10^?8 g/mL (signal‐to‐noise ratio = 3). Application of the present method to the analysis of FA in human urine proved feasible. Copyright © 2009 John Wiley & Sons, Ltd.     

Change of the Protein p53 Electrochemical Signal According to its Structural Form – Quick and Sensitive Distinguishing of Native, Denatured, and Aggregated Form of the “Guardian of the Genome”

Presence of mutated and/or structurally modified (e.g., denatured, aggregated) protein p53 form is associated with several disorders such as Alzheimer’s disease, Parkinson’s disease, prion diseases, and many types of tumours. The aim of this work was to distinguish native, denatured and aggregated form of full-length p53 by flow injection analysis coupled with electrochemical detector (FIA-ED). Firstly FIA-ED method used for protein native form determination was optimized (detection limit 45.8 amol per 5 μl injection; 3×S/N). In addition the technique was applied to identify p53 structural forms (denatured and aggregated). It was found out that denatured form provides about three times higher electrochemical response (protein structure unfolding, approach of more electroactive centers – aminoacid residues – towards electrode surface) in comparison with native form. On the other hand, aggregated form offers lower response (steric eclipse of electroactive protein parts) when compared with the signal of native form. The obtained data show that we are not only able to sensitively determine native, denatured, and aggregated structural forms of p53 protein but also to distinguish them.