毛细管等电聚焦/毛细管无胶筛分电泳二维蛋白质分离平台的构建

设计并制作了一种新型的中空纤维接口,以此为核心构建了毛细管等电聚焦(CLEF)／毛细管无胶筛分(CNGE)电泳二维蛋白质分离技术平台,实现了将二维凝胶电泳从平板转移到毛细管中。利用该二维分离平台,对血红蛋白样品进行了高效、快速分离分析,验证了该二维分离平台的可行性及分离效能。实验结果表明：二维分离系统总的峰个数和分离效能都比各自一维分离系统有较大提高。     

毛细管电泳进样技术新进展

评述了毛细管电泳进样技术新成果。对直接在线进样，二维分离体系中毛细管电泳分离的增样，相关毛细管电泳增样，超微量样品及单个分子的进样，近端进样，双向进样和高温下的进样装置的应用状况作了介绍。     

无机阴离子的毛细管电泳分析

本文研究了常见无机阴离子的毛细管电泳规律，考察了分离电压、电解质熔液组成、浓度、ｐＨ值等对分离的影响，建立了高效、快速的无机阴离子毛细管电泳分析方法。在选定的实验条件下，各种常见无机阴离子在５ｍｉｎ内达到完全分离，对Ｂｒ＾－和Ｃｌ＾－的分离柱效每米可达７６万理论板数。迁移时间的相对标准偏差小于１％，峰面积的相对标准偏差小于５％，各离子的最低检测浓度为０．０５￣０．５μｇ／ｍｌ。     

毛细管电泳有效分离测定甜菊糖苷新方法的研究

本文采用100％二甲基甲酰胺（DMF）作为样品的溶剂,以Tris－硼砂作为分离甜菊糖苷的缓冲体系,探讨了进样量、柱温、电压、Tris-硼砂缓冲液浓度以及pH值等条件对甜菊糖苷分离结果的影响。通过优化各种分离条件,实现了毛细管电泳高效分离测定甜菊糖苷。     

毛细管等电聚焦/加压毛细管电色谱二维分离体系在多肽分离中的应用

以毛细管等电聚焦(cIEF)为第一维分离模式,以反相加压毛细管电色谱(pCEC)为第二维分离模式,开展离线二维色谱分离研究,并对复杂肽段进行分离.羟丙基纤维紊(HPC)涂层的毛细管用于cIEF分离,对6种标准蛋白质的平均分离柱效约为31万.在毛细管末端引进电隔离槽,方便了第一维样品的收集.在加电6 kV下,第二维pCE...     

无死体积蚀刻多孔膜接口的制作及其在二维毛细管电泳分离系统中的应用

介绍了一种在毛细管柱上原位腐蚀而成的多孔膜接口的制作方法,并用该接口构建了一类毛细管电泳二维分离技术平台。柱上原位腐蚀刻成的多孔膜接口具有零死体积、制作过程简易、成本低廉、耐用、柱间切换便捷等优点,特别适合作为基于毛细管柱的二维及多维电泳联用中的接口,是目前二维及多维毛细管柱联用中一类较为新型、实用、理想的接口。以鹿茸冻干粉可溶物样品为例,验证了该接口在二维毛细管电泳联用系统中的可行性和分离效能。实验结果表明：鹿茸冻干粉可溶物整个二维分离分析的时间在1 h内完成,二维分离系统的分辨率和总峰容量都比一维的高。     

色氨酸、半胱氨酸和酪氨酸的高效毛细管电泳分析

用自组装的高效毛细管电泳安培检测装置,对具有常规电活性的色氨酸,半胱氨酸和酪氨酸进行了分离分析条件的研究,采用重力进样方式,进样高度25cm,进样时间15S,在分离毛细管长70cm,内径25um,的电泳装置上,10mmol/L K2HPO4-H3PO4缓冲液（PH10．5）,20KV分离高压,＋1．05V（vs SCE)检测电位的条件下,对酸解毛发中得到的混合氨基酸中的色氨酸,半胱氨酸和酪氨酸进行了分离测定,结果令人满意。     

微透析中空纤维膜接口的制作及其在二维毛细管电泳联用中的应用

提出了一种微透析中空纤维膜的制备方法,并用该微透析中空纤维膜作为接口构建了一套新型的二维毛细管电泳分离技术平台。中空纤维膜接口具有制作过程简易、传质效率高、柱间切换方便、分析速度快等优点,它是目前二维及多维电泳联用中一种较为新型、方便、理想的接口。以血红蛋白样品为例,通过实验验证了该接口在二维毛细管电泳联用系统中的可行性和分离效能。实验结果表明:整个血红蛋白样品二维分离分析的时间在1 h内完成,二维分离系统的分辨率和总峰容量都比各自一维的高。     

用高效毛细管电泳安培法检测扑热息痛及其水解物对氨基酚

采用高效毛细管电泳安培法对扑热息痛及其水解物对氨基酚的测定进行了研究。优化选择了工作电极、检测电位、缓冲沉沦浓度等实验参数,并探讨了进样时间与峰电流的关系。结果表明：在优化的实验条件下扑热息痛和对氨基酚在６ｍｉｎ内可以得到较好的分离,检出限分别为０．５ｕｍｏｌ／Ｌ和１．０ｒｍｏｌ／Ｌ。该法适合药物生产过程的质量控制和临床检测。     

毛细管电泳检测神经细胞凋亡

建立了毛细管电泳检测凋亡细胞DNA片段的方法.以DNA相对分子质量标准品为溶质,考察了分离条件(电压、进样时间、温度、聚合物浓度)对分离的影响.在优化条件下,利用毛细管无胶筛分电泳对缺氧缺血过程中不同时间点神经PC12细胞DNA片段进行了分析,并与流式细胞仪结果比较,研究缺氧缺血时胶质细胞凋亡过程.     

Comprehensive two-dimensional chromatography and capillary electrophoresis coupled with tandem time-of-flight mass spectrometry for high-speed proteome analysis

A comprehensive two-dimensional capillary liquid chromatography and capillary zone electrophoresis system coupled with tandem matrix assisted laser desorption/ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-TOF-MS) proteomics analyzer is presented. Protein/peptide samples were separated by capillary high-performance liquid chromatography (cHPLC). The effluents from cHPLC (the first dimension) were continuously transferred into capillary zone electrophoresis (CZE, the second dimension) through a novel valve-free hydrodynamic sampling interface. The CZE effluents were mixed with alpha-cyano-4-hydroxycinnamic acid (CHCA) matrix sheath flow via CE-MALDI interface, and then directly deposited on the MALDI target at a 3 s time-interval for further MS analysis. The high efficiency of the overall system was demonstrated by analysis of proteins in D20 (human hepatocellular carcinoma model in nude mice with high metastatic potential) liver cancer tissue. More than 300 proteins were identified, which proved the system potential for high-throughput analysis and application in proteomics.     

使用含离子涂层柱的毛细管电泳和毛细管电色谱的研究进展

 毛细管电色谱是近年发展起来的高效、高选择性的微分离技术。与一般的毛细管电泳和使用ODS反相填料的毛细管电色谱相比 ,含离子涂层柱的毛细管电泳和毛细管电色谱能提供较大且可控的电渗流 ,便于拓宽分离对象 ,优化分离条件。对使用含离子涂层柱的毛细管电泳和电色谱的特点、发展和应用状况进行了综述。     

毛细管等电聚焦-毛细管区带电泳二维蛋白质分离平台的构建及其性能

通常采用二维聚丙烯酰胺凝胶电泳 ( 2 D- PAGE)分析组织或细胞的全蛋白质 [1] ,但难与质谱 ( MS)直接联用 .用高效液相色谱 ( HPLC)和毛细管电泳 ( CE)分离分析蛋白质和多肽的一维分离模式的分辨率和峰容量有限 .多维柱联用技术比一维分离有更高的分辨率和峰容量 [2 ] ,便于和 MS直接联用 [3,4 ] ,易于实现自动化 .目前 ,有关 2 D- CE的报道相对较少 [5～ 7] ,我们初步实现了将 2 D- PAGE由平板转移到毛细管中[6 ,7] ,但凝胶柱的制作烦琐 ,存在交叉污染 ,不能与 MS直接联用 .本文用微透析中空纤维膜为接口构建了毛细管等电聚焦 ( …     

平面二维毛细管电泳初探

在石英单晶表面制成短形截面的毛细管柱上进行了电泳实验。由于矩形柱比国形住有更大散热侧面积且石英单晶的导热性能远远优于熔融石英,所以可施加较高的场强,不仅提高了住效,而且缩短了分离时间。两个相交的通道之间形成自然连接,可实现二维分离,并消除了死体积。     

高效毛细管电泳电化学检测器的研制

本文提出了一种用于高效毛细管电泳的新型安培电化学检测器设计，使用Ｎａｆｉｏｎ溶液制作的ＨＰＣＥ／ＥＤ接口，可有效地隔开两化学系统的干扰，且不引入附加体积，经对有机酚类化合物的胶束电动毛细管色谱分离与电化学检测知，该系统性能优良，对对苯二酚的检出限为３０ａｍｏｌ。     

Capillary zone electrophoresis: Effect of physical parameters on separation efficiency and quantitation

Separation efficiency in capillary zone electrophoresis was examined as a function of capillary length and diameter as well as solute concentration for a borosilicate glass capillary/phosphate buffer system. Electroosmotic flow coefficients in borosilicate, fused silica, and teflon capillaries were measured over the useful operating pH range of 3–8 at constant applied power (0.333 W). Sample introduction by a simple, low-dispersion electromigration method was found to be a reliable procedure for solute quantitation.     

电填充毛细管色谱柱性能的评价

报道了一种毛细管色谱柱的电填充技术,并进行了柱性能的评价研究。实验表明新的填充方法确保了色谱柱的柱效和重现性。     

用于高效毛细管电泳的电化学检测器

阐述了高效毛细管电泳电化学检测器（包括电导、电势和安培检测）的研究现状，重点是检测器的研制及接口的制作技术。对各种电化学检测器的应用情况也进行了总结。展望了高效毛细管电泳电化学检测的发展前景。     

Two-dimensional separation system of coupling capillary liquid chromatography to capillary electrophoresis for analysis of Escherichia coli metabolites

A two-dimensional (2-D) separation system of coupling chromatography to electrophoresis was developed for profiling Escherichia coli metabolites. Capillary liquid chromatography (LC) with a monolithic silica-octadecyl silica column (500 x 0.2 mm ID) was used as the first dimension, from which the effluent fractions were further analyzed by capillary electrophoresis (CE) acting as the second dimension. Field-enhanced stacking was selectively employed as a concentration strategy to interface the two dimensions, which proved to be beneficial for the detection of metabolites. An artificial sample containing 118 standards, some of which lack chromophores or have weak UV absorbance, was used to optimize the 2-D separation system. Under the optimum conditions, 63 components in the artificial sample having absorbance at 254 nm could be well resolved and detected. The utility of the system was demonstrated by comprehensive analysis of E. coli metabolites. Comparing with the previous 2-D separation system we published in Anal. Chem. 2004, 76, 1419-1428, using a longer monolithic column in the first dimension improved the separation efficiency and offered the possibility of increasing the injection volume without compromising the separation efficiency. In the second dimension, field-enhanced stacking was used to improve the concentration sensitivity of the metabolites, and more metabolites in E. coli cell extract were detected and identified using the developed 2-D separation system. In addition, preliminary investigation for future CE-mass spectrometry coupling was also made in the study by using volatile buffers in the capillary LC and CE techniques.