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.     

Determination of cefotaxime and desacetylcefotaxime in cerebrospinal fluid by solid-phase extraction and high-performance liquid chromatography

A microfluidic analytical system for the separation and detection of organic peroxides, based on a microchip capillary electrophoresis device with an integrated amperometric detector, was developed. The new microsystem relies on the reductive detection of both organic acid peroxides and hydroperoxides at -700 mV (vs. Ag wire/AgCl). Factors influencing the separation and detection processes were examined and optimized. The integrated microsystem offers rapid measurements (within 130 s) of these organic-peroxide compounds, down to micromolar levels. A highly stable response for repetitive injections (RSD 0.35-3.12%; n = 12) reflects the negligible electrode passivation. Such a "lab-on-a-chip" device should be attractive for on-site analysis of organic peroxides, as desired for environmental screening and industrial monitoring.     

Observations on a dropping-mercury electrochemical detector for flow-injection analysis and HPLC

An electrochemical detector cell having a dropping-mercury working electrode and an Ag/ AgCl reference electrode is examined with reference to applications in flow-injection analysis and high-performance liquid chromatography. The working electrode, fed by a horizontal capillary, is capable of delivering mercury at fast dropping rates which can be varied by a control device in the cell. Examples of application to inorganic and organic electroreducible species are considered and advantages and limitations relative to other electrochemical detector devices are discussed.     

Carbon-nanotube/copper composite electrodes for capillary electrophoresis microchip detection of carbohydrates

The preparation of carbon nanotube (CNT)/copper composite electrodes, based on co-mixing CNT and Cu powders within mineral oil, is described. The new composite electrode is used for improved amperometric detection of carbohydrates following their capillary electrophoresis (CE) microchip separations. The CNT/Cu composite electrode detector displays enhanced sensitivity compared to detectors based on copper or CNT alone. The marked catalytic action of the CNT/Cu composite material permits effective low potential (+0.5 V vs. Ag/AgCl) amperometric detection, and is coupled to the renewability, bulk modification and versatility advantages of composite electrodes. The CNT/Cu composite surface also leads to a greater resistance to surface fouling compared to that observed at the copper electrode. Factors affecting the electrocatalytic activity and the CE microchip detection are examined and optimized. The CNT/Cu composite electrode is also shown to be useful for the detection of amino acids as indicated from preliminary results. While the present work has focused on the enhanced CE microchip detection of carbohydrates and amino acids, the CNT/metal-composite electrode route should benefit the detection of other important groups of analytes.     

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.     

HPLC Urinary Organic Acid Profiling: Role of the Ultraviolet and Amperometric Detectors

Abstract

The profiling of urinary organic acids is an important aspect of the diagnosis of inborn metabolic disorders. The carboxylic acids of interest may contain additional functional moities such as phenyl, hydroxyl, oxo, etc. The state-of-the-art method of organic acid analysis is by GC/MS. Prior to GC/MS analysis, the carboxylic acids must be isolated from urine by extraction or ion exchange chromatography and made volatile by derivatization. This is a lengthy procedure that does not lend itself to rapid analysis. We have developed a rapid procedure for the profiling of urinary α, B-unsaturated, aromatic and α-ketocarboxylic acids.

Urine containing an internal standard, 3-hydroxy-4-methoxy-benzyl alcohol, is filtered through a 0.3 um Millipore filter and injected on to an HPX-87 organic acid HPLC column (Bio-Rad). The mobile phase, 4.5 mN H₂SO₄, is passed through the column at 0.8 ml/min. Detection is effected by an UV detector at 200 nm in series but upstream from an electrochemical detector with a glassy carbon working electrode at +1.15V vs. an Ag/AgCl reference electrode. At this electrode potential, phenolic, methoxyphenyl, eneolic and α-ketocarboxylic acids are oxidized and can be electrochemically detected with a glassy carbon electrode.     

Carbon paste-based electrochemical detectors for microchip capillary electrophoresis/electrochemistry

The first reported use of a carbon paste electrochemical detector for microchip capillary electrophoresis (CE) is described. Poly(dimethylsiloxane) (PDMS)-based microchip CE devices were constructed by reversibly sealing a PDMS layer containing separation and injection channels to a separate PDMS layer that contained carbon paste working electrodes. End-channel amperometric detection with a single electrode was used to detect amino acids derivatized with naphthalene dicarboxaldehyde. Two electrodes were placed in series for dual electrode detection. This approach was demonstrated for the detection of copper(II) peptide complexes. A major advantage of carbon paste is that catalysts can be easily incorporated into the electrode. Carbon paste that was chemically modified with cobalt phthalocyanine was used for the detection of thiols following a CE separation. These devices illustrate the potential for an easily constructed microchip CE system with a carbon-based detector that exhibits adjustable selectivity.     

Determination of taurine in human tear fluid by capillary electrophoresis with indirect amperometric detection based on electrogenerated bromine

A new method for the determination of taurine was developed based on indirect amperometric detection after capillary electrophoresis. A serial dual‐electrode detector comprising an on column Pt film electrode (upstream electrode) and an end column Pt microdisk electrode (downstream electrode) was utilized to conduct the indirect amperometric detection. Bromide is oxidized to bromine at upstream electrode and reduced back to bromide at downstream electrode. Since taurine can react with bromine quantitatively and rapidly, its concentration can therefore be determined by the decrease of the current for bromine reduction at the downstream electrode. Principal experimental parameters governing the analytical performance were investigated and optimized. Under the optimal conditions, taurine can be baseline separated from interfering amino acids and the detection limit of 0.18 μM was obtained with a linear correlation coefficient of 0.999 over the concentration range of 0.5–60 μM. The developed method has been successfully applied in the determination of taurine in human tear fluid. The taurine level obtained was in good agreement with previous reports and recoveries for taurine spiked ranged from 92–95% with relative standard deviations within 4.6%, demonstrating the reliability of the developed method in the determination of taurine in human tear fluid.     

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。     

Fabrication of graphene/poly(methyl methacrylate) composite electrode for capillary electrophoretic determination of bioactive constituents in Herba Geranii

This report describes the development and application of a novel graphene/poly(methyl methacrylate) composite electrode as a sensitive amperometric detector of capillary electrophoresis. The composite electrode was fabricated on the basis of the in situ polymerization of a mixture of graphene and prepolymerized methyl methacrylate in the microchannel of a piece of fused silica capillary under heat. SEM, XRD and FT-IR offered insights into the nature of the composite. The results indicated that graphenes were well dispersed in the composite to form an interconnected conducting network. The performance of this unique graphene-based detector has been demonstrated by separating and detecting seven naturally occurring phenolic compounds in Herba Geranii in combination with capillary electrophoresis. The graphene-based detector offered significantly lower operating potentials, substantially enhanced signal-to-noise characteristics, and lower expense of operation. The simplicity and significant performance exhibited by the graphene/poly(methyl methacrylate) composite electrode also indicate great promise for microchip CE, flowing injection analysis, and other microfluidic analysis systems.     

Operational parameters of voltammetric high-performance liquid chromatographic detectors with copper electrodes and application to a determination of some fodder biofactors

Two versions of an amperometric detector with a copper working electrode have been constructed and tested for high-performance liquid chromatography (HPLC). The performance of the detectors was studied using selected amino acids. The dependence of the detector response on the mobile phase flow-rate was studied in the range common in both macro- and microcolumn HPLC (5 microliter/min to 1.0 ml/min). It has been found that the detection sensitivity generally increases with decreasing flow-rate, i.e., the detector response is governed by the rate of the complexation reaction between the cupric ions and the solutes. This fact makes amperometric detection with a copper electrode especially useful for microcolumn separations. For all 20 amino acids studied, calibration curve parameters and detection limits have been determined; the latter vary from 0.4 to 18 ng in the injected volume. The amino acids can also be sensitively detected in a medium of 0.1-1.0 M ammonia, which is promising for the use of strong anion exchangers in amino acid separations. Choline can also be detected at a copper electrode, with a detection limit of 40 ng. An HPLC method with amperometric detection at a copper electrode has been developed for the determination of lysine, methionine and choline in fodder biofactors.     

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。     

Miniaturized thin-layer radial flow cell with interdigitated ring-shaped microarray electrode used as amperometric detector for capillary electrophoresis

A chip-type thin-layer radial flow cell was developed as an amperometric detector for capillary electrophoresis. We fabricated a carbon film-based interdigitated ring-shaped array (IDRA) microelectrode with a 2 microm bandwidth and an almost 1 microm gap on a glass plate and used it as a working electrode. A fused-silica capillary was arranged above the IDRA electrode using a guide hole drilled through the acryl plate that formed the flow cell lid. A flow channel for use in connecting the outlet capillary was also fabricated in the acryl plate. We characterized the analytical performance of the IDRA electrode in the microchip flow cell in terms of linear concentration range, sensitivity and concentration detection limit. We achieved a collection efficiency and catechol redox cycle at the IDRA microelectrode of 65% and 1.71, respectively, and thus a high sensitivity and low detection limit of 392.9 pA/microM and 15 nM for dopamine hydrochloride. We examined the reproducibility of the detector and found that the run-to-run and detector-to-detector relative standard deviations were both less than 10%.     

End-column electrochemical detection for inorganic and organic species in high-voltage capillary electrophoresis

The electronics and construction for an end-column ultramicroelectrode (3–10 μm) detection system that permits the use of medium-sized capillaries (25 μm I.D.) without appreciable effects from the potential field at the end of the capillary. Normal peak-to-peak noise over 10 s was 0.01–0.1 pA. The background noise observed for a 200 × μm carbon-fiber electrode placed either 180 μm within a 25-μm capillary or at a point 500 μm away from the capillary was essentially the same. A study of detector response as a function of the position of the electrode has shown that accurate location of the electrode is important for sensitive and reproducible detection. These studies also showed that differences between the density of the electrolyte existing the capillary and the electrolyte in the detection cell could cause anomalous electrode response depending on the location of the electrode relative to the end of the capillary. Application of a carbon fiber or an Hg film electrode gave detection limits (twice the peak-to-peak noise over 10 s) of 2 · 10⁻⁸ mol/l for Pb²⁺, 1 · 10^{− 5} mol/l for NO₂⁻ and 5 · 10⁻¹⁰ mol/l for catechol.     

毛细管电泳脉冲伏安电化学分离检测胺类化合物

电化学检测高效毛细管电泳自 80年代末问世以来 ,已迅速发展为一种重要的分析方法 ,但是具有许多优点的现代电分析化学方法在毛细管电泳检测中的应用还不多 .扫描伏安电化学检测是继安培检测方法之后应用较多的一种方法 ,它除具有安培检测方法的优点外 ,还能得到被测物质的电流随电压变化的伏安特性 [1～ 5 ] ,但由于在电极电位的扫描过程中会产生较大的充电电流 ,进而影响检测的灵敏度和检出限 .脉冲安培检测可获得较高的灵敏度和低的检出限 ,并减少电极的污染 ,因而被应用于高效液相色谱 [6,7] 和毛细管电泳 [8,9] 中检测有机化合物 .目前…     

Multi-purpose capillary electrophoresis system with concentration gradient detection

A robust, inexpensive and versatile capillary electrophoresis (CE) system for routine and rapid analysis is reported, which consists of a rugged cartridge holding a 20-mum i.d. 15-cm long capillary, and an inexpensive, universal and sensitive concentration gradient detector. The design of the cartridge simplifies the sample introduction process and makes it possible to perform many separation modes, including moving boundary capillary electrophoresis (MBCE), capillary zone electrophoresis (CZE), capillary isotachophoresis (CITP) and capillary isoelectric focusing (CIEF), on the same system. This arrangement provides more information about a sample's components since analytes can be separated by different modes performed on the same CE system. The detector only consists of a low-power HeNe laser, or laser diode, and a photodiode position sensor. Amino acids and proteins of 10(-6)-10(-3)M concentration can be separated by different capillary electrophoretic modes, and detected directly by the detector. The universal detector shows particularly good sensitivity when applied to CE separation modes having self-concentration and focusing effects. Femtomoles of proteins were separated and detected with CIEF. In addition, a short and narrow capillary allows use of high electrical fields which facilitate rapid separations. Four amino acids at millimolar concentrations were fully separated and detected in less than 80 sec by the MBCE mode when a high electric field was applied. The physical size of the whole system is much smaller than that of conventional CE instruments with UV absorbance or fluorescence detector.     

Development and Characterization of a Disposable Electrochemical Detector Arrangement for Capillary Electrophoresis Prepared by Thermal Lamination

A disposable electrochemical detector arrangement for capillary electrophoresis has been developed. The detection system consists of an assembly of a fused silica capillary and a microfiber electrode which are fixed in a proper position by sealing this configuration with the help of thermal lamination. The detector design is easy to prepare and inexpensive. Therefore it is used as a disposable device which can be replaced by a new one if any degradation of the performance characteristics occurs. The parameters influencing the detection performance were studied and optimized using nonaqueous capillary electrophoresis and ferrocene derivatives as model compounds. The practical utility was demonstrated by studying the separation and detection of water‐resistant dye compounds.     

Microchip Capillary Electrophoresis with a Single-Wall Carbon Nanotube/Gold Electrochemical Detector for Determination of Aminophenols and Neurotransmitters

A new SWCNT modified gold detector for microchip capillary electrophoresis–electrochemistry is described. SWCNT modified gold electrode displays greatly improved sensitivity and separation resolution compared to bare gold electrode, reflecting the electrocatalytic activity of SWCNT. The SWCNT/Au electrode exhibits low background noise levels. Parameters such as separation voltage and detection potential of the microchip electrophoresis–electrochemistry with SWCNT modified gold electrode were optimized.     

Facile preparation of carbon nanotube/poly(ethyl 2-cyanoacrylate) composite electrode by water-vapor-initiated polymerization for enhanced amperometric detection

A carbon nanotube/poly(ethyl 2-cyanoacrylate) (CNT/PECA) composite electrode was developed for enhanced amperometric detection. The composite electrode was fabricated on the basis of water-vapor-initiated polymerization of a mixture of CNTs and ethyl 2-cyanoacrylate in the bore of a piece of fused silica capillary. The morphology and structure of the composite were investigated by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and thermogravimetric analysis. The results indicate that the CNTs were well dispersed and embedded throughout the PECA matrix to form an interconnected CNT network. The analytical performance of this unique CNT-based detector has been demonstrated by separating and detecting six flavones in combination with capillary electrophoresis. The advantages of the CNT/PECA composite detector include lower operating potential, higher sensitivity, low expense of fabrication, satisfactory resistance to surface fouling, and enhanced stability; these properties indicate great promise for a wide range of applications.     

Graphene/poly(ethylene‐co‐vinyl acetate) composite electrode fabricated by melt compounding for capillary electrophoretic determination of flavones in Cacumen platycladi

In this report, a graphene/poly(ethylene‐co‐vinyl acetate) composite electrode was fabricated by melt compounding for the amperometric detection of capillary electrophoresis. The composite electrode was fabricated by packing a mixture of graphene and melted poly(ethylene‐co‐vinyl acetate) in a piece of fused silica capillary under heat. The structure of the composite was investigated by scanning electron microscopy and Fourier transform infrared spectroscopy. The results indicated that graphene sheets were well dispersed in the composite to form an interconnected conducting network. The performance of this unique graphene‐based detector has been demonstrated by separating and detecting rutin, quercitrin, kaempferol, and quercetin in Cacumen platycladi in combination with capillary electrophoresis. The four flavones have been well separated within 9 min in a 50‐cm‐long capillary at a separation voltage of 12 kV using a 50 mM sodium borate buffer (pH 9.2). The graphene‐based detector offered significantly lower operating potentials, substantially enhanced signal‐to‐noise characteristics, lower expense of operation, high resistance to surface fouling, and enhanced stability. It showed long‐term stability and repeatability with relative standard deviations of <5% for the peak current (n = 15).