LIQUID CHROMATOGRAPHY OF BIOLOGICAL SUBSTANCES

说明：（申请人摘要）本提案旨在进一步 扩大现代液相色谱的范围，以满足 医学和生命科学对分析分离方法的需求 高速度和分辨率。 主要重点是开发和使用 新型填充毛细管柱，以探索毛细管的全部潜力 电色谱法 该液相色谱技术采用 填充毛细管柱和高电场驱动移动的相 并承诺实现迄今为止仅能实现的高峰容量 在高分辨率气相色谱和毛细管区带电泳中， 分别对挥发性和带电样品组分进行分析。 一 主要的研究工作将是阐明物理化学 洗脱液流动的性质、产生和控制的基础， 并开发一系列特别定制的新型柱 以满足毛细管电色谱中的特殊要求。 此外，还介绍了这种新技术在分离和 分析具有生物学意义的复杂混合物， 类胡萝卜素通过肽和蛋白质转化为糖缀合物， 将探索多糖。 拟议的研究将进行 沿着几条密切相关的路线。 在研究了 在上一个项目期间的色谱过程中， 发展到估计优惠的相互作用参数是有用的 在生物物质的表征中， 蛋白质，从数据很容易获得的疏水相互作用 层析 组合化学中的高通量筛选 需要快速的分析方法，并继续早期的研究， 高温高效液相色谱填充毛细管柱研究 增强的 毛细管电泳的优点将进一步得到 在零度以下的温度下检查， 肽、蛋白质和碳水化合物，以及各种 这类物质的聚集形式。 新的应用前景 环糊精在毛细管电色谱和毛细管中的应用 肽、蛋白质和其他生物聚合物的电泳将是 研究了 所有的研究建议将大大受益于熔融石英 基于正在开发的新型毛细管柱。 其中包括在 现场形成的流体不可渗透的聚合物内管或多孔整体 填料，每种填料具有适合于特定用途的表面性质。 应用程序.

DESCRIPTION: (applicant's abstract) This proposal is aimed at further expanding the scope of modern liquid chromatography in a major way to meet the needs in medicine and life sciences for analytical separation methods of high speed and resolution. The main focus is on the development and use of novel packed capillary columns to explore the full potential of capillary electrochromatography. This liquid chromatographic technique employs a packed capillary column and high electric field to drive the mobile phase and promises the realization of high peak capacities attainable so far only in high resolution gas chromatography and capillary zone electrophoresis in the analysis of volatile and charged sample components respectively. A major research effort will be made to shed light on the physico-chemical underpinning of the nature, generation and control of eluent flow by electrosmosis and to develop a family of novel columns especially tailored to meet the particular requirements in capillary electrochromatography. Furthermore, the application of this new technique to the separation and analysis of complex mixtures of biological significance varying from carotenoids through peptides and proteins to glycoconjugates and polysaccharides will be explored. The proposed research will be carried along several closely related lines. Following studies on the energetics of the chromatographic process in the last project period, a method will be developed to estimate preferential interaction parameters which are useful in the characterization of biological substances including therapeutical proteins, from data readily obtained in hydrophobic interaction chromatography. High throughput screening in combinatorial chemistry requires rapid analytical methods and in continuation of earlier "studies, the research" on high temperature HPLC with packed capillary columns will be augmented. The advantages of capillary electrophoresis will be further examined at subzero temperatures with readily interconverting conformers of peptides, proteins and carbohydrates, and in the separation of various agglomerated forms of such substances. Novel applications for the promising use of cyclodextrins in capillary electrochromatography and capillary electrophoresis of peptides, proteins and other biopolymers will be investigated. All studies proposed will greatly benefit from fused silica based novel capillary columns that are under development. They include in situ formed fluid impervious polymeric innertube or porous monolithic packing, each having surface properties appropriate for the particular application.