毛细管电泳在生物大分子分离中的应用

本文回顾了近年来毛细管电泳在蛋白质和DNA分析中的应用及一些新的技术的进展情况,并介绍蛋白质-药物结合物的发现和分离,医学检验中对复杂样品的检测,在核酸分析中的荧光染色剂、筛分剂以及芯片毛细管电泳等方面的内容．     

毛细管电泳非接触电导检测器的研制

非接触电导检测法已经成为毛细管电泳芯片中一种通用的物质分离检测方法．本文提出了一种微型化的电容耦合非接触电导分析系统,采用新颖的三明治电极结构减小了电极之间的寄生电容．这种电极结构能够有效地减小电极的长度,克服了非接触电导检测中电极易于折断的缺点．当采用电极宽度1mm、电极间距1mm、MES／His缓冲溶液浓度为20mmol／L和激励电压20Vpp、90kHz的条件时,检测器具有最好的分离效果．在最佳的分离效果,实现了1mmol／L的K^＋和Mg^2＋混合无机阳离子的分离检测．     

区带毛细管电泳法分析黑色签字 笔墨水字迹相对形成时间

讨论了优化电泳条件建立区带毛细管电泳法分析黑色签字笔字迹墨水的分析方法,采用２０mmol桙L四 硼酸钠,pH８畅５,未涂层熔融石英毛细管内径１００μm,有效柱长４０cm,１５kV分离电压,光二极管阵列检测器 （PDA）,检测波长１９０～６００nm的电泳分离条件,对不同种类黑色签字笔字迹墨水提取液进行了分析;利用毛细 管电泳PDA检测器在可见光波长下检测出的黑色签字墨水中不同染料相对含量的变化关系,研究了字迹相对形 成时间问题,得出较好的变化规律。     

小肽BCEOC衍生物的非水毛细管电泳分离研究

以1,2-苯并-3,4二氢咔唑-9-乙基氯甲酸酯(BCEOC)作为柱前衍生试剂,采用非水毛细管电泳对衍生小肽进行分离。在甲醇乙腈混合液为溶剂,乙酸-乙酸铵为缓冲体系,20℃,28kV,235nm二极管阵列检测(DAD)条件下,采用非水毛细管电泳模式,考察并优化小肽衍生物的分离条件,实现四种小肽衍生物的基线分离。     

表面贴附微透镜阵列的OLED诱导荧光检测系统

为进一步减小光源的体积、简化光路系统结构，消除毛细管电泳芯片诱导荧光检测系统中激发光源的反射光和杂散光的干扰，设计并建立了垂直层叠结构的有机发光二极管诱导荧光检测系统。针对有机发光二极管器件发光强度较低的问题，采用在器件的玻璃基底表面贴附微透镜阵列薄膜的方法，提高了入射光的强度。在对CCD积分时间、PDMS微透镜直径及两偏振片间的偏振角度等参数进行优化之后，利用检测系统实现了罗丹明B样品溶液的毛细管电泳分离。     

芯片毛细管电泳技术的应用与进展

介绍了目前芯片毛细管电泳的主要分离技术:芯片毛细管区带电泳、芯片毛细管凝胶电泳、芯片胶束电动色谱、芯片毛细管电色谱及多相层流无膜扩散分离.芯片毛细管电泳的研究已经取得了很大的进展,研制出了多种微型化、集成化的芯片,多种分离技术在芯片上的开发应用,显示出芯片毛细管电泳技术在众多分析领域会有广阔的发展前景.     

基于水平分置线性双阵列的超声全聚焦成像方法在粗晶材料检测中的应用

针对粗晶材料超声检测信噪比低的问题,提出了一种水平分置线性双阵列超声成像方法。将两个线阵超声换能器沿直线水平分置在待检区域表面两侧,用收发分离的信号采集模式,一侧激发,另一侧记录各通道数据,进行聚焦成像。相比单阵列和同位置双线阵检测,文中的方法有效地减少了背向散射信号对缺陷信号的干扰,提高了成像信噪比。在粗晶铜质试块上的成像实验结果表明,当缺陷距离阵列较近时,文中的方法优于单阵列和同位置双线阵方法,成像信噪比提高约5～10 dB;当缺陷距离阵列较远时,单阵列模式和同位置双线阵检测方法失效,但文中的方法依然可以识别缺陷。文中的研究为粗晶材料的超声检测提供了一种可行的方案。     

毛细管电泳水相电解质溶液的选择研究

在毛细管电泳中选择合适的电解质溶液可以改变分离的选择性,提高分离度和分离效率,因此对电解质溶液的选择和优化是非常必要的。对毛细管电泳水相电解质的选择与优化进行综述。     

非水毛细管电泳溶剂系统的选择与优化

在非水毛细管电泳中选择合适的溶剂可以改变分离的选择性,提高分离度和分离效率,因此对溶剂系统的选择和优化是非常必要的。就此对非水毛细管电泳溶剂系统的选择与优化进行综述。     

土壤苯酚测定中毛细管电泳—二极管阵列检测法的应用

本研究通过采用毛细管电泳—二极管阵列检测法测定土壤中的苯酚，经过对不同性质的土壤样品进行分析及质控实验，结果证明该方法检测饿回收率为98．4％，说明该方法，精密度和准确度均符合质量分析求，成功地应用于土壤中苯酚的测定。     

Laser-induced fluorescence detection by liquid core waveguiding applied to DNA sequencing by capillary electrophoresis

A new laser-induced fluorescence detector for capillary electrophoresis (CE) is described. The detector is based on transverse illumination and collection of the emitted fluorescent light via total internal reflection along the separation capillary. The capillary is coated with a low refractive index fluoropolymer and serves as a liquid core waveguide (LCW). The emitted light is detected end-on with a CCD camera at the capillary exit. The observed detection limit for fluorescein is 2.7 pM (550 ymol) in the continuous-flow mode and 62 fM in the CE mode. The detector is applied to DNA sequencing. One-color G sequencing is performed with single-base resolution and signal-to-noise ratio approximately 250 for peaks around 500 bases. The signal-to-noise ratio is approximately 50 for peaks around 950 bases. Full four-color DNA sequencing is also demonstrated. The high sensitivity of the detector is suggested to partly be due to the efficient rejection of scattered laser light in the LCW. The concept should be highly suitable for capillary array detection.     

Comparison of Hadamard transform and signal-averaged detection for microchannel electrophoresis

A Hadamard transform (HT) detection method for microchip capillary electrophoresis with laser-induced fluorescence and a charge-coupled device (CCD) is described and compared to signal-averaged detection. A low-noise CCD camera is used to image a section of a separation channel where each camera pixel can be thought of as a unique detector. For signal averaging, electropherograms corresponding to individual pixels can be averaged for improved S/N. HT detection is performed on each pixel electropherogram to generate a contour plot electropherogram. The multiple injections required for HT provides an enhancement at the cost of longer times for the pseudorandom injection sequences. A short sample injection length of 0.25 s is used to reduce the overall analysis time and improve sensitivity compared to previously published results. An injection sequence is performed on the microchip that is based on a cyclic S-matrix of 513 elements that generates an 8-fold improvement in S/N compared to a single injection. This spatially resolved HT detection method is also capable of performing a multicomponent separation. Signal-averaged HT and single-injection data are compared to experimental HT and single-injection results. The unique capabilities of each method are described.     

Capillary electrophoresis with an integrated on-capillary tubular detector based on a carbon sol-gel-derived platform

An integrated on-capillary tubular electrochemical detector for capillary electrophoresis systems has been fabricated based on sol-gel technique. It consists of a sol-gel carbon composite tubular electrode attached permanently onto the outlet of the separation capillary. The device greatly eases the setting up of capillary electrophoresis with electrochemical detection (CEEC) as it makes possible electrode/capillary alignment without the aid of a micromanipulator since this integrated unit can be simply immersed in the CE separation buffer in an ordinary three-electrode stationary cell. To improve analytical performance of the integrated unit, the external wall of the exit capillary was etched with HF after the polyimide coating of the capillary had been removed. Influences of the working electrode length and the wall thickness at the outlet of capillary on the separation efficiency and amperometric sensitivity were assessed and optimized. The practical applicability of this configuration is demonstrated with the detection of both catecholamines and carbohydrates. The advantages, namely, versatility, convenience, ease of operation, and low-cost, of the new design combined with an excellent performance lead to high stability and low detection limits.     

Improving the compatibility of contact conductivity detection with microchip electrophoresis using a bubble cell

A new approach for improving the compatibility between contact conductivity detection and microchip electrophoresis was developed. Contact conductivity has traditionally been limited by the interaction of the separation voltage with the detection electrodes because the applied field creates a voltage difference between the electrodes, leading to unwanted electrochemical reactions. To minimize the voltage drop between the conductivity electrodes and therefore improve compatibility, a novel bubble cell detection zone was designed. The bubble cell permitted higher separation field strengths (600 V/cm) and reduced background noise by minimizing unwanted electrochemical reactions. The impact of the bubble cell on separation efficiency was measured by imaging fluorescein during electrophoresis. A bubble cell four times as wide as the separation channel led to a decrease of only 3% in separation efficiency at the point of detection. Increasing the bubble cell width caused larger decreases in separation efficiency, and a 4-fold expansion provided the best compromise between loss of separation efficiency and maintaining higher field strengths. A commercial chromatography conductivity detector (Dionex CD20) was used to evaluate the performance of contact conductivity detection with the bubble cell. Mass detection limits (S/N = 3) were as low as 89 +/- 9 amol, providing concentration detection limits as low as 71 +/- 7 nM with gated injection. The linear range was measured to be greater than 2 orders of magnitude, from 1.3 to 600 microM for sulfamate. The bubble cell improves the compatibility and applicability of contact conductivity detection in microchip electrophoresis, and similar designs may have broader application in electrochemical detection as the expanded detection zone provides increased electrode surface area and reduced analyte velocity in addition to the reduction of separation field effects.     

侧壁四电极电容耦合非接触电导检测器(英文)

研制了一种集成于硅基电泳芯片分离沟道末端侧壁的新型四电极电容耦合非接触电导检测器.研究了该电导检测器的等效模型,对等效电路模型中的参数进行了公式推导,并讨论了影响电导检测响应灵敏度的相关因素.采用深刻蚀及离子注入加工技术制得了用于电导检测的立体电极.制作了基于锁相放大原理的信号处理电路,对该电导检测的频率响应及灵敏度进行了测试分析.实验结果表明,当激励信号频率为300 kHz时,该电导检测器具有最佳线性度;不同浓度Na+溶液响应电压差值为5 mV;检测限达到10-8mol/L;且成功实现了Na+和Li+混合无机阳离子的电泳分离在线检测.     

Fourier transform capillary electrophoresis with laminar-flow gated pressure injection

Fourier transform capillary electrophoresis (FTCE) was developed as a method to improve signal-to-noise ratio (S/N) and resolution in capillary electrophoresis (CE) separation. In FTCE, multiple simultaneous CE separations were performed in the same channel system and interrogated using a single-point detector. To illustrate experimentally the improvement offered by FTCE in S/N ratio and resolution, we carried out a modest number (five) of multiple injections and separations. We show even with this small number of separations, S/N increased by a factor of 2.9, and theoretical plate height improved by a factor of more than 30. We demonstrated this technique with laser-induced fluorescence detection, but a wide variety of detection methods are compatible with FTCE.     

Real-Time Imaging through Optical Fiber Array-Assisted Laser-Induced Fluorescence of Capillary Electrophoretic Enantiomer Separations

An advanced detection system based on laser-induced fluorescence imaging for capillary electrophoresis (CE) is presented. An optical fiber array was constructed for collection and transportation of the emitted fluorescent light to the charge-coupled device (CCD) camera. The fiber array makes the setup compact compared with a setup where the capillary is imaged through a camera objective. The imaging detector captures the sample zones in motion during the migration through the capillary. This allows unique studies on dynamic events otherwise unrevealed. During the study, unexplained nonlinear migration behavior was revealed. Enantiomer separations of dansylated amino acids using cyclodextrins, imaged between 1.5 and 12 cm of a 28-cm-long 50-μm i.d. capillary, were used for evaluation of the system. Comparing the optical fiber array with a camera lens system, the signal-to-noise-ratio (S/N) was 10 times higher. This is due to a combination of both higher signal and lower noise levels. To improve the S/N ratio further, a computer program for signal processing was designed. Using dichlorofluorescein, a concentration limit of detection (CLOD) of 350 pM was achieved and improved 10 times to 35 pM with computer postprocessing using 79 images. This is equal to 400 zeptomole for a 3-mm-long sample zone in a 50-μm i.d. capillary.     

High-performance genetic analysis on microfabricated capillary array electrophoresis plastic chips fabricated by injection molding

We have developed a novel technique for mass production of microfabricated capillary array electrophoresis (mu-CAE) plastic chips for high-speed, high-throughput genetic analysis. The mu-CAE chips, containing 10 individual separation channels of 50-microm width, 50-microm depth, and a 100-microm lane-to-lane spacing at the detection region and a sacrificial channel network, were fabricated on a poly(methyl methacrylate) substrate by injection molding and then bonded manually using a pressure-sensitive sealing tape within several seconds at room temperature. The conditions for injection molding and bonding were carefully characterized to yield mu-CAE chips with well-defined channel and injection structures. A CCD camera equipped with an image intensifier was used to monitor simultaneously the separation in a 10-channel array with laser-induced fluorescence detection. High-performance electrophoretic separations of phiX174 HaeIII DNA restriction fragments and PCR products related to the human beta-globin gene and SP-B gene (the surfactant protein B) have been demonstrated on mu-CAE plastic chips using a methylcellulose sieving matrix in individual channels. The current work demonstrated greatly simplified the fabrication process as well as a detection scheme for mu-CAE chips and will bring the low-cost mass production and application of mu-CAE plastic chips for genetic analysis.