New Chromatographic Technologies for Resolving Carbohydrate Isomers

解析碳水化合物异构体的新色谱技术

• 批准号: 8985005
• 负责人: Milos Novotny
• 金额: $ 31.6万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8985005
• 关键词: Address; Adsorption; Area; Biocompatible Materials; Biological; Biological Markers; Blood capillaries; Buffers; Caliber; Carbohydrates; Carbon; Chemicals; Chemistry; Chromatography; Collaborations; Complex; Complex Mixtures; Detection; Detergents; Development; Diagnostic Neoplasm Staging; Disease; Ensure; Equipment; Evaluation; Geometry; Glycobiology; Glycoconjugates; Glycopeptides; Glycoproteins; Heart; In Situ; Indiana; Isomerism; Kinetics; Laboratories; Lead; Lectin; Length; Liquid substance; Mass Spectrum Analysis; Materials Testing; Measurement; Medical; Methodology; Methods; Microspheres; Modeling; Modification; Monosaccharides; Morphology; North Carolina; Oligonucleotides; Oligosaccharides; Particle Size; Peptides; Performance; Phase; Physiological; Polymers; Polysaccharides; Procedures; Protein Glycosylation; Proteomics; Pump; Research; Research Personnel; Resolution; Salts; Science; Scientist; Silanes; Silicon Dioxide; Solvents; Source; Specialist; Structure; Surface; System; Techniques; Technology; Testing; Time; Tissues; Tube; Ultrasonics; Universities; Variant; Work; base; capillary; capillary liquid chromatography; carboxylate; human disease; instrument; interest; medical specialties; meetings; metabolomics; particle; physical property; pressure; professor; public health relevance; silane; solute; sugar; tandem mass spectrometry

 DESCRIPTION (provided by applicant): Separation of complex mixtures from biological sources has been highly dependent on advances in microcolumn (capillary) liquid chromatography (LC). When used in combination with biomolecular mass spectrometry (MS), capillary LC columns have been at the heart of key "omics" technological approaches including glycoproteomic and glycomic profiling of biological fluids and tissues in search for disease biomarkers. While typical columns used in the analytical practice utilize 5 µm-sized particles packed inside capillary tubes, more kinetically progressive column types can now be tightly packed with particles within 1.0- 1.5 µm range. At the expense of higher inlet pressures and the needed time for optimizing column packing technologies, the capillaries packed with such small particles can clearly outperform the more conventional columns used in today's practice of proteomics and metabolomics (as it has been already shown with reversed-phase separations). However, the separation problems of the glycoscience need different column materials effectively retaining relatively hydrophilic carbohydrates. Highly promising new column materials have been recently developed at Indiana University and preliminarily tested to meet such needs: (1) macroporous silica microspheres, which are prepared by ultrasonic spray pyrolysis (USP) using inorganic salts as a removable pore template; and (2) macroporous carbon microspheres, employing organic carboxylate salts, whose USP yields morphologically well-defined entities. This proposal deals with appropriate modifications of the particle surfaces for the benefits of adsorption chromatography and hydrophilic interaction chromatography (HILIC) developments and toward providing thus far unachieved total resolution of numerous glycan isomers. This is to be achieved through the optimum combination of column selectivity and kinetic performance. The proposal involves a combination of expertise by the P.I., a bioanalytical chemist and a separation science specialist, and the co-I. specializing in materials chemistry. The study will also be significantly aided by a distinguished collaborator at the University of North Carolina and two other collaborators with significant expertise in carbohydrate synthesis. Aim 1 concerns modification of macroporous silica microparticles for retention of polar glycoconjugate solutes in the HILIC separation mode. The USP-derived macroporous silica particles will be packed into capillaries of different lengths and diameters and evaluated under different regimes of ultrahigh pressure and different mobile phases. Special consideration will be given to biologically important fucosylated and sialylated isomeric structures. After packing silica materials, the column will be treated in situ to generate polar surface structures. Aim 2 will deal with packing different types of particles, specifically carbonaceous materials. They will be subjected to simila evaluations as in Aim 1. The proposed research will address one of the urgent needs of contemporary glycobiology and biomedical field.

 描述（通过应用程序提供）：从生物来源分离复杂的混合物已经高度依赖于Microsumn（毛细管）液相色谱法（LC）的进步。当与生物分子质谱法（MS）结合使用时，毛细血管LC柱已成为关键“ OMICS”技术方法的核心，包括生物液和组织的糖蛋白质组学和糖核分析，以寻找疾病生物标志物。虽然分析实践中使用的典型色谱柱利用了毛细管内部包装的5 µm尺寸的颗粒，但现在可以在1.0-1.5 µm范围内将更多动力学渐进柱类型紧紧地用颗粒紧密包装。以较高的入口压力和优化柱填料技术的必要时间，填充了如此小的颗粒的毛细血管可以显然胜过当今蛋白质组学和代谢组学实践中使用的更传统的柱子（因为已经显示出与相反的相位分离显示的那样）。但是，糖科学的分离问题需要不同的柱材料有效地保留相对亲水性的碳氢化物。最近在印第安纳大学开发了高度有希望的新柱材料，并初步测试以满足此类需求：（1）通过使用无机盐作为可移动的孔隙模板，使用无机盐使用超声波喷雾热解（USP）制备的大孔二氧化硅微球； （2）使用有机羧酸盐盐的大孔微球，其USP产生形态上定义明确的实体。该建议介绍了颗粒表面的适当修改，以使吸附色谱和亲水相互作用色谱（HILIC）开发的益处，并旨在提供迄今为止迄今未能达到的众多聚糖异构体的总分辨率。这是通过柱选择性和动力学性能的最佳组合来实现的。该提案涉及P.I.的专业知识，生物分析化学家和分离科学专家以及Co-I的组合。专门研究材料化学。北卡罗来纳大学的杰出合作者和 另外两名在碳质合成方面具有重要专业知识的合作者。 AIM 1涉及对大孔二氧化硅微粒的修改，以在Hilic分离模式下保留极性糖缀合物溶质。 USP衍生的大型二氧化硅颗粒将被包装成不同长度和直径的毛细血管，并在不同的超高压力和不同移动相的不同状态下进行评估。将对具有生物学上重要的岩藻糖基化和溶解的异构体结构进行特殊考虑。包装二氧化硅材料后，将原位处理柱子以产生极性表面结构。 AIM 2将处理包装不同类型的颗粒，特别是碳质材料。与AIM 1一样，他们将接受Simila评估。拟议的研究将解决当代糖生物学和生物医学领域的紧急需求之一。