Construction and evaluation of a novel end-column amperometric detection system for electrophoresis microchips

A set of integrated end-column amperometric detection system has been developed,onto which an electrophoresis microchip can be conveniently integrated.Finely machined by a piece of transparent organic glass,the system consists of an electrophoresis microchip platform and an amperometric detection reservoir,in which the microchip can be fixed onto the platform by microchip grooves and with stainless steel fixture.Each detection electrode can be directly fixed in the amperometric detection reservoir by screws...     

新型芯片电泳柱端安培检测系统的构建与评价

自行设计开发了一套便于与电泳芯片集成的一体式柱端安培检测池系统．该系统由整块透明有机玻璃精密加工而成,包括电泳芯片支架和安培检测池两部分,芯片可通过芯片插槽和不锈钢夹具固定在芯片支架上,各种检测用电极可直接通过螺母固定在安培检测池中．以100μmol／L的DA为模式分析物,分别采用直径为100、300和500μm的铂金圆盘电极与表观直径为240μm的碳纤维电极作为工作电极均在该装置上实现了良好组装和高灵敏检测．采用碳纤维工作电极对该系统的检测参数进行了优化．测试结果表明该系统在电化学清洗程序下连续六次测定100μmol／L多巴胺的峰电流相对标准偏差为3．2％,保留时间相对标准偏差为0．5％,DA的检测限为0．4μmol／L（按照S／N=3计）．该系统体积小巧,测试稳定,检测灵敏度较高,工作电极更换方便,适合作为芯片电泳柱端安培检测通用平台．     

微流控芯片技术在蛋白质分析中的应用进展

近年来,微流控芯片技术取得了显著的发展。随着微电子及微机械等制作技术的不断进步,高通量、高效、快速、低成本的微流控分析芯片在蛋白质和多肽分析方面获得了令人瞩目的成果。本文主要介绍了微流控芯片在蛋白质组学分析研究的应用和发展。引用文献52篇。     

Manipulation of the Electroosmotic Flow in Glass und PMMA Microchips with Respect to Specific Enzymatic Glucose Determinations

The determination of glucose in microfluidic chips made of glass or PMMA was used as a model for the combination of an enzymatic reaction with the separation of compounds. It was based on the enzymatic oxidation of glucose and the amperometric detection of hydrogen peroxide. Real samples frequently contain compounds, such as ascorbic acid, which may interfere with quantitative glucose determinations. Thus, electrophoretic separation of specific from unspecific signals was envisaged by applying electric fields which are also used to control the flow of liquid via electroosmotic effects. Surface charge densities of the capillaries influence the electroosmotic flow (EOF). They are dependent on the chip material and on the adsorption of components from the background electrolyte. Reversal of the EOF after addition of cetyltrimethylammonium bromide (CTAB) and an increase in EOF after addition of sodium dodecylsulfate (SDS) were observed at lower surfactant concentrations with the PMMA chips rather than with the glass chips. For both chip materials these concentrations were below the critical micelle concentration. Effective separation of H₂O₂ and ascorbic acid was achieved with low CTAB concentrations, which lead to a reduction, but not to a reversal of the EOF. Reversal of the EOF by higher CTAB concentrations or the increase in cathodic EOF by SDS accelerated ascorbic acid transportation and reduced the differences in migration times. Thus, for the specific determination of glucose, glucose oxidase was added together with low CTAB concentrations to the background electrolyte. This avoided interference from ascorbic acid, and data obtained from the analysis of fruit juices showed a good correlation to data obtained from a reference method.     

An integrated plastic microchip for enhancing electrophoretic separation using tunable pressure-driven backflows

Microfluidic CE (MCE) is an effective solution for rapid and sensitive determination of multiple analytes. Herein, a dynamic coated cyclic olefin copolymer microchip was developed having an on-chip micropump for fluid velocity adjusting in electrophoretic separations. This micropump was fabricated by constructing a polyacrylamide gel membrane at one channel terminal. Once applying electric field across the membrane, a pressure-driven flow generated automatically to balance the electroosmotic flow (EOF) mismatch at the channel-membrane interface. The influence of gel precursor concentration and operating voltages on the fluid velocity was carefully evaluated. Moreover, the highly integration of injection, separation, and pumping units of the MCE system minimized the dead volume and provides satisfied column efficiency. Experiments showed that by adjusting of pumping voltage reduced the fluid velocity by a factor of 6, resulting six- and threefold resolving power enhancements of rhodamine dye mixture and amino acid mixture, respectively. Furthermore, the developed MCE method was applied for rhodamines and amino acids quantitation in food and cosmetics, with standard addition recoveries of 87.3–106.9% and 89.9–117.4%, respectively. These results were also confirmed by standard HPLC method, revealing the application potential in fast and onsite analysis of complex samples.     

Sensitivity enhancing injection from a sample reservoir and channel interface in microchip electrophoresis

The stacking of a cationic analyte (i.e., rhodamine B) at the interface between a sample reservoir and channel in a microchip electrophoresis device is described for the first time. Stacking at negative polarity was by micelle to solvent stacking where the dye was prepared in a micellar solution (5 mM sodium dodecyl sulfate in 25 mM phosphoric acid, pH 2.5) and the channel was filled with high methanol content background solution (70% methanol in 50 mM phosphoric acid, pH 2.5). The injection of the stacked dye into the channel was by simple reversal of the voltage polarity with the sample solution and background solution at the anodic and cathodic reservoirs of the straight channel, respectively. The enrichment of rhodamine B at the interface and injection of the stacked dye into the channel was clearly visualized using an inverted fluorescence microscope. A notable sensitivity enhancement factor of up to 150 was achieved after 2 min at 1 kV of micelle to solvent stacking. The proposed technique will be useful as a concentration step for analyte mixtures in simple and classical cross‐channel microchip electrophoresis devices or for the controlled delivery of enriched reagents or analytes as narrow plugs in advanced microchip electrophoresis devices.     

External-cavity birefringence feedback effects of microchip Nd:YAG laser and its application in angle measurement

External-cavity birefringence feedback effects of the microchip Nd:YAG laser are presented. When a birefringence element is placed in the external feedback cavity of the laser, two orthogonally polarized laser beams with a phase difference are output. The phase difference is twice as large as the phase retardation in the external cavity along the two orthogonal directions. The variable extra-cavity birefringence, caused by rotation of the external-cavity birefringence element, results in tunable phase difference between the two orthogonally polarized beams. This means that the roll angle information has been translated to phase difference of two output laser beams. A theoretical analysis based on the Fabry--Perot cavity equivalent model and refractive index ellipsoid is presented, which is in good agreement with the experimental results. This phenomenon has potential applications for roll angle measurement.     

基于微腔型电极的半乳糖生物传感器

采用微机械加工技术制作了具有三维结构的微腔型传感器芯片（腔深３００μｍ,铂工作电极１．２＊１．２＾ｍｍ＾－２。作者对微腔型电极进行了电化学特征的表征。并通过明胶包埋法将半乳糖氧化酶固定在电极中,构成微腔型半乳糖传感器。     

芯片毛细管电泳程控电源的研制

设计并制作了一种小型的、可与PC机相联的程序控制电源，其中有两路输出电压，用于芯片毛细管电泳的分离和进样；而且，可程序控制输出电压和运行时间，并在PC机上以图形方式显示是泳电流。该电源适用于芯片毛细管电泳的过程监控。     

Assembly-controlled biocompatible interface on a microchip: strategy to highly efficient proteolysis

A biocompatible interface was constructed on a microchip by using the layer-by-layer (LBL) assembly of charged polysaccharides incorporating proteases for highly efficient proteolysis. The controlled assembly of natural polyelectrolytes and the enzyme-adsorption step were monitored by using a quartz-crystal microbalance and atomic force microscopy (AFM). Such a multilayer-assembled membrane provides a biocompatible interconnected network with high enzyme-loading capacity. The maximum digestion rate of the adsorbed trypsin in a microchannel was significantly accelerated to 1600 mM min(-1) microg(-1), compared with the tryptic digestion in solution. Based on the Langmuir isotherm model, the thermodynamic constant of adsorption K was calculated to be 1.6 x 10(5) M(-1) and the maximum adsorption loading Gammamax was 3.6 x 10(-6) mol m(-2), 30 times more than a monolayer of trypsin on the native surface. The tunable interface containing trypsin was employed to construct a microchip reactor for digestion of femtomoles of proteins and the produced peptides were analyzed by MALDI-TOF mass spectroscopy. The efficient on-chip proteolysis was obtained within a few seconds, and the identification of biological samples was feasible.