Combination of amperometric detector & UV detector for capillary electrophoresis

A new electrochemical cell assembly with the combination of UV and amperometric detector (AD) based on their complementarity was described. A Nafion tubing junction was used to decouple the high voltage from the separation capillary in the rear of on-column UV detector. In this mode, the electroactive and inert compounds could be detected by UV and AD at the same time. Aromatic amines were determined with the UV and the end-column AD detection to evaluate the performances of the cell assembly. Such an improved electrochemical detector could match the capillary with different diameters. By simple adjustment of the screws, the positioning of the working electrode and the detection capillary was easily gained without microscope. It is also very easy to assemble and disassemble the working electrode when needed.     

一种基于双工作电极-双通道的毛细管电泳电化学检测系统

报道了一种双工作电极-双通道毛细管电泳电化学检测系统，实现电导和安培 同时检测或者安培与安培检测联用，使两种方法相互补充，发挥各自的优势。其中 ，工作电极与检测池的制作工艺简单，操作简便，通过不锈钢针管和毛细管作为套 管，无需三维微调装置即可简单实现双工作电极的准确放置及分离毛细管与工作电 极的准确对接，并根据分析体系的需要采用不同类型的工作电极和检测器；同时采 用复式滤波电路解决了不同检测器之间的电场叠加对输出信号的干扰问题。采用该 装置可以同时检测复杂体系中的电活性和惰性物质，或同时测定只能氧化或只能还 原的物质，还可以对具有氧化还原性质的物质进行纯度的确证。将该装置应用于实 际样品的测定，节约了分析时间，提高了分析速度，扩大了检测范围，结果令人满 意。     

The In-cell iR Compensation Using a Four-electrode System

It is well known that the iR compensation is very important in electrochemistry, especially in fast, ultra-fast and transient voltammetry for kinetic and mechanistic studies. The modern design of potentiostat is usually of the three-electrode system, in w…     

A New Dual-electrode and Multi-channel Electrochemical Detection System for Capillary Electrophoresis

A new type of dual-electrode and multi-channel electrochemical detection technology for capillary electrophoresis is described in this paper.Two detectors(the amperometric detector and the conductometric detector) or two conductometric detectors are connected to the same capillary electrophoresis system.The whole system possesses the advantages of the two electrochemical detectors including sparing time,improving the analytical speed and expanding the sample range.The working electrode and detector cell are handled easily.The system was applied to sample detection with satisfactory results.     

Determination of morphine and its major metabolite, morphine-3-glucuronide, in blood by high-performance liquid chromatography with electrochemical detection

The construction of an electrochemical detector flow cell for high-performance liquid chromatography from stainless steel, which also serves as the auxiliary electrode, is described. Glassy carbon is used for the working electrode in a novel configuration based on a hybrid of the wall-jet and thin-layer arrangements. Application of the detector to the determination of morphine in small blood samples is described. The metabolite, morphine-3-glucuronide, is also determined by hydrolysis with beta-glucoronidase prior to extraction. The detector is capable of detecting less than 1 ng of morphine injected.     

Dual Electrochemical Detection of Biogenic Amine Metabolites in Micro High-Performance Liquid Chromatography

Abstract

The dual electrochemical detectors for ordinary and micro high-performance liquid chromatography were briefly reviewed.

The electrochemical behaviors of biogenic amine metabolites were studied by cyclic semi-differential and semi-integral voltammetry with a glassy carbon working electrode. It was found that the electrochemical reactions of many biogenic amine metabolites are quasi-reversible. The dual electrochemical detector based on thin-layer electrolytic cell with two working electrodes (anode and cathode) in series configuration was tested for selective detection of biogenic amine metabolites on their electrochemical quasi-reversibility. The detector was successfully utilized for the simultaneous determination of 3, 4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindole-3-acetic acid in human urine directly injected by micro high-performance liquid chromatography.     

毛细管电泳多道电化学检测器的研制及其应用

报道了毛细管电泳多道电化学检测器的研制及其应用，安培检测器和电导检测器并联在同一毛细管电泳检测系统中，在同一缓冲体系，同一工作电极下对同一复杂的分析体系同时进行电导和安培检测；整个装置综合了电导检测和安培检测两种检测器的优点，性能优良，造价低廉，对实际样品的检测取得了令人满意的结果。     

Determination of oestriol in pregnancy urine by normal-phase high-performance liquid chromatography with electrochemical detection

A method for the determination of oestriol in pregnancy urine by normal-phase high-performance liquid chromatography with electrochemical detection is described. A large-volume wall-jet cell with an Ag-Ag+ reference electrode was used as the detector system. The limit of detection obtained is comparable to that of electrochemical detection following reversed-phase liquid chromatography. One of the advantages of electrochemical detection with normal-phase systems is that adsorption problems are minimized.     

一种铂网栅薄层光谱电化学池的制作及表征

以铂网栅电极作工作电极,设计制作了一种应用于紫外-可见光谱电化学研究的新型光透薄层电化学池（OTTLE）,并以铁氰化钾-亚铁氰化钾体系对此光谱化学池进行了表征。铁氰化钾-亚铁氰化钾体系在该池中有良好的循环伏安特性,并可获得良好的紫外-可见光谱。测得的克式量电位E^0,电子转移数n与文献结果一致。     

Swept-potential oxidative detection in flow streams

A swept-potential electrochemical detector, operated in the oxidative staircase voltammetric metric mode, is demonstrated for the high-performance liquid chromatography of a mixture of catecholamines. Voltammetric limits of detection are approximately 30 pg or 1 nM and chromatographic limits of detection are approximately 250 pg. The use of a platinum working electrode in a wall-jet cell configuration, with potential pulses for cleaning and activation between each sweep, results in a cell that has maintained a constant response for over a year without mechanical refinishing of the electrode surface.     

Reductive Mode Thin-Layer Amperometric Detector for Liquid Chromatography

Abstract

An electrochemical detector for liquid chromatography is described based on the use of a mercury film electrode in a thin-layer cell with the auxiliary electrode placed across the channel from the working electrode. Considerations relevant to optimizing performance of the detector are described including removal of dissolved oxygen from the sample and mobile phase. Detection limits are reported for a number of reducible organic compounds of interest in environmental and biomedical research. Detection limits at the picomole level are readily achieved for easily reduced nitro compounds. Typical applications are illustrated for the determination of parathion in field runoff water and chloramphenicol in human plasma.     

An electrochemical detector cell for open tubular liquid chromatography and capillary electrophoresis

Summary An electrochemical detector cell has been developed for micro-flow separation systems (OTLC, CE). The cell contains two electrodes, a disk-shaped working electrode made from a carbon fiber bundle, and a tubular Ag/AgCl quasi-reference electrode. The effective cell volume and the coulometric yield have been determined, for different electrode diameters and at different flow rates, in an OTLC system. An effective cell volume of less than 1 nl was observed. The applicability of the cell was demonstrated with the detection of OPA-derivatized amino acids. For use in CE, the cell was equipped with an additional compartment, housing a semi-permeable joint for the decoupling of the high electric field used for the electrophoretic separation. Results are shown on the determination of catecholamines by CE with electrochemical detection. Detection limits with both OTLC and CE were well below 1 fmole.     

A Modified Z-Type Flow-through Cell for Optical, Electrochemical, and Optoelectrochemical Flow Injection Analysis Measurements

The design and construction of a modified Z-type flow-through cell that can be used for optical, electrochemical, and optoelectrochemical measurements are reported. In optical analysis, the proposed flow cell is placed in the light path of a spectrophotometer and worked as a commercial Z-type optical flow cell. In electrochemical analysis, the working and auxiliary electrodes are two platinum sheets placed at both ends of the cell cavity; the cell cavity is enclosed by two glass windows. The reference electrode is a saturated calomel electrode placed downstream in an overflow tube. The performance of the proposed electrochemical flow cell was studied by analysis of hexacyanoferrate solution as an electroactive species. In optoelectrochemical analysis, the proposed flow cell is operated optically and electrochemically, simultaneously. The flow cell is placed in the light path of a spectrophotometer, constant potential is applied to the electrodes, and the optical and electrochemical signals are recorded simultaneously by the use of two recorders.     

Microchip reversed-phase liquid chromatography with packed column and electrochemical flow cell using polystyrene/poly(dimethylsiloxane)

A microchip pressure-driven liquid chromatography (LC) with a packed column and an electrochemical flow cell has been developed by using polystyrene (PS) and poly(dimethylsiloxane) (PDMS). The cylindrical separation column with packed octadecyl silica particles was fabricated in the PS substrate. The three electrode system (working, reference, and counter electrode) for amperometric detection was fabricated onto the PS substrate, using the Au deposition, photolithography, and chemical etching. The detector flow cell was formed by sealing the electrode system with a PDMS chip containing a channel. In this flow cell, the effect of working electrode width (in the direction of flow) on chromatographic parameters, such as peak width and peak resolution were studied in electrode width ranging 50-5,000 microm. The effect of electrode width on sensitivity (current intensity, current density, and S/N ratio) was also examined. The sensitivity was discussed by simulating the concentration profile generated around the working electrode. The effects of the column packing size and the column size on the separation efficiency were examined. In this study, a good separation of three catechins was successfully achieved and the detection limits for (+)-catechin, epicatechin, and epigallocatechin gallate were 350, 450, and 160 nM, respectively.     

Liquid Chromatographic Determination of Glutathione with Electrochemically Pretreated Glassy Carbon Electrode

Abstract

Simple electrochemical pretreatment of a glassy carbon electrode used as a working electrode in an electrochemical detector has been found to enhance the analytical capability of the detector for the determination of glutathione by lowering the required operating potential as well as by increasing the maximum oxidation current. The electrochemical pretreatment of the electrode was made in a 0.2 M phosphate buffer (KH₂PO₄-KOH, pH 6.5) at +1.9 V vs. Ag/AgCl for 2 min. The minimum detectable quantity of glutathione was found to be about 100 pg, when eluate from a reversed-phase column was amperometrically monitored by aid of the detector with the electrochemically pretreated glassy carbon electrode. Amount of glutathione in a rat lens was determined by the chromatographic method developed in this study.     

Disposable Amperometric Sensors for Thiols with Special Reference to Glutathione

The antioxidant ‘reduced glutathione’ tripeptide is conventionally called glutathione (GSH). The oxidized form is a sulfur‐sulfur linked compound, known as glutathione disulfide (GSSG). Glutathione is an essential cofactor for antioxidant enzymes; it provides protection also for the mitochondria against endogenous oxygen radicals. The ratio of these two forms can act as a marker for oxidative stress. The majority of the methods available for estimation of both the forms of glutathione are based on colorimetric and electrochemical assays. In this study, electrochemical sensors were developed for the estimation of both GSH and GSSG. Two different types of transducers were used: i) screen‐printed three‐electrode disposable sensor (SPE) containing carbon working electrode, carbon counter electrode and silver/silver chloride reference electrode; ii) three‐electrode disposable system (CDE) consisting of three copper electrodes. 5,5′‐dithiobis(2‐nitrobenzoic acid) (DTNB) was used as detector element for estimation of total reduced thiol content. The enzyme glutathione reductase along with a co‐enzyme reduced nicotinamide adenine dinucleotide phosphate was used to estimate GSSG. By combining the two methods GSH can also be estimated. The detector elements were immobilized on the working electrodes of the sensors by bulk polymerization of acrylamide. The responses were observed amperometrically. The detection limit for thiol (GSH) was less than 0.6 ppm when DTNB was used, whereas for GSSG it was less than 0.1 ppm.     

A Self‐contained Two‐electrode Nanosensor for Electrochemical Analysis in Aqueous Microenvironments

We describe the development, fabrication, and characterization of a novel two‐electrode nanosensor contained within the tip of a needle‐like probe. This sensor consists of two, vertically aligned, carbon structures which function as individual electrodes. One of the carbon structures was modified by silver electrodeposition and chlorination to enable it to function as a pseudo‐reference electrode. Performance of this pseudo‐reference electrode was found to be comparable to that of commercially available Ag/AgCl reference electrodes. The unmodified carbon structure was employed as a working electrode versus the silver‐plated carbon structure to form a two‐electrode sensor capable of characterizing redox‐active analytes. The nanosensor was demonstrated to be capable of electrochemically characterizing the redox behavior of para‐aminophenol (PAP) in both bulk solutions and microenvironments. PAP was also measured in cell lysate to show that the nanosensor can detect small concentrations of analyte in heterogenous environments. As the working and reference electrodes are contained within a single nanoprobe, there was no requirement to position external electrodes within the electrochemical cell enabling analysis within very small domains. Due to the low‐cost manufacturing process, this nanoprobe has the potential to become a unique and widely accessible tool for the electrochemical characterization of microenvironments.     

The use of pulsed amperometry combined with ion-exclusion chromatography for the simultaneous analysis of ascorbic acid and sulfite

Initial attempts to monitor ascorbic acid and sulfite, in a beer matrix, by combining ion-exclusion chromatography with a pulsed amperometric detector using a single applied voltage to the platinum working electrode, were unsuccessful. Alternatively, good chromatograms for the separation of the two antioxidants were achieved utilizing a standard, amperometric cell. However, remarkably superior results were observed when this standard cell was operated in a pulsed mode and cleaning cycles were continually applied throughout the analysis. The working electrode stability and precision have been examined. Preliminary spike recovery data indicate acceptable accuracy for the method. Comparisons of this method to standard reference methods are currently ongoing.     

Noise and Drift Phenomena in Amperometric and Coulometric Detectors for HPLC and FIA

Abstract

Noise and drift phenomena in electrochemical detectors with solid electrodes are discussed. A relationship between the capacity of the working electrode and the noise of the detector is demonstrated in three different ways, using direct correlation of noise with capacity, time correlation functions and electrical simulation of the cell properties. Conclusions are drawn with respect to the prospects of various measures to improve the detection limit.     

Sheath-flow electrochemical detection of amino acids with a copper wire electrode in capillary electrophoresis

Here, we report the detection of native amino acids using a sheath-flow electrochemical detector with a working electrode made of copper wire. A separation capillary that was inserted into a platinum tube in the detector acted as a grounded electrode for electrophoresis and as a flow channel for sheath liquid. Sheath liquid flowed outside the capillary to support the transport of the separated analytes to the working electrode for electrochemical detection. The copper wire electrode was aligned at the outlet of the capillary in a wall-jet configuration. Amino acids injected into the capillary were separated following elution from the end of the capillary and detection by the copper electrode. Three kinds of copper electrodes with different diameters-50, 125, and 300 μm-were examined to investigate the effect of the electrode diameter on sensitivity. The peak widths of the analytes were independent of the diameter of the working electrode, while the 300-μm electrode led to a decrease in the signal-to-noise ratio compared with the 50- and 125-μm electrodes, which showed no significant difference. The flow rate of the sheath liquid was also varied to optimize the detection conditions. The limits of detection for amino acids ranged from 4.4 to 27 μM under optimal conditions.