LIQUID CHROMATOGRAPHY OF BIOLOGICAL SUBSTANCES

描述：(申请人摘要)本提案旨在进一步 扩大现代液相色谱领域的主要途径是满足 分析分离方法在医学和生命科学中的需求 高速度和高分辨率。主要关注的是开发和使用 新型填充毛细管柱充分发挥毛细管潜力 电色谱。该液相色谱技术采用 填充毛细管柱和高电场驱动流动相 并承诺实现目前为止仅能达到的高峰值容量 在高分辨气相色谱和毛细管区带电泳法中 挥发性和带电样品组分的分析。一个 将进行重大研究工作，以阐明这种物理化学 支持洗脱液流动的性质、产生和控制 电雾化和开发一系列特别定制的新型色谱柱 以满足毛细管电色谱的特殊要求。 此外，这一新技术在分离和分离中的应用 对具有生物学意义的复杂混合物的分析 类胡萝卜素通过多肽和蛋白质转化为糖共轭化合物和 将对多糖进行探索。将进行拟议的研究 沿着几条密切相关的路线。在此基础上进行的能量学研究如下 在最后一个项目期的层析过程中，会有一种方法 开发用于估计有用的优先相互作用参数 在生物物质的表征中，包括治疗性物质 蛋白质，来自疏水相互作用中容易获得的数据 层析法。组合化学中的高通量筛选 需要快速的分析方法，并继续进行早期的“研究”， 填充式毛细管柱高温高效液相色谱的研究 增加了。毛细管电泳法的优势将进一步 在零度以下的温度下检查，很容易相互转化为 多肽、蛋白质和碳水化合物的分离，以及各种 这类物质的凝聚形式。前景看好的新应用 环糊精在毛细管电色谱和毛细管中的应用 多肽、蛋白质和其他生物聚合物的电泳法 调查过了。所有提出的研究都将极大地受益于熔融二氧化硅 基于正在开发中的新型毛细管柱。它们包括在 原位成型流体防渗聚合物内管或多孔整体 包装，每个包装都具有适合于特定产品的表面特性 申请。

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.