A Raman detector for Field-Flow Fractionation for advanced in vitro characterization of Macromolecules

用于场流分级分离的拉曼检测器，用于大分子的高级体外表征

• 批准号: 424650380
• 负责人: Professor Dr. Helmut Cölfen (†)
• 金额: --
• 依托单位: Arbeitsgruppe Physikalische Chemie (AG Cölfen)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/424650380?language=en
• 关键词: Raman; detector; Field; Flow; Fractionation

This project aims to set up an on-line Raman detection system for asymmetrical-flow field flow fractionation (AF4). AF4 is a separation method for macromolecules, which also allows the determination of distributions of the diffusion coefficient resp. the hydrodynamic size over a range from 1 nm to 100 micro meters. Current detectors yield distributions of the molar mass and the relative sample concentrations. Also UV-Vis or fluorescence spectra for each component in complex mixtures can be measured. The proposed Raman detection introduces the possibility to gain information about the chemical nature of the sample. By this way, fractionated samples in complex mixtures could be distinguished solely by chemical groups without requiring any labelling as it is necessary for the distinction of many samples using UV-Vis and fluorescence detectors. Comparable methods to our proposal are only available under enormous financial expenses and with a strictly limited number of usable elution solvents. Even the single components of completely unknown samples could be fractionated and their chemical groups be identified in the Raman spectra. The Raman detector shall be tested with commercially available standard proteins and protein complexes with small molecules, nucleic acids or carbohydrates. Also, stabilizer molecules on core-shell nanoparticles shall be investigated as their chemical composition in situ should deliver valuable information for the optimization of nanoparticle synthesis, which cannot be obtained properly by existing methods.

本计画旨在建立一套非对称流场流体分馏（AF 4）的在线拉曼侦测系统。AF 4是一种用于大分子的分离方法，它还允许测定扩散系数的分布。流体动力学尺寸在1纳米至100微米的范围内。电流检测器产生摩尔质量和相对样品浓度的分布。还可以测量复杂混合物中每种组分的紫外-可见或荧光光谱。所提出的拉曼检测引入了获得关于样品的化学性质的信息的可能性。通过这种方式，复杂混合物中的分级样品可以仅通过化学基团来区分，而不需要任何标记，因为使用UV-Vis和荧光检测器区分许多样品是必要的。与我们的提议类似的方法只有在巨大的财务费用和严格有限数量的可用洗脱溶剂下才可用。即使是完全未知的样品的单一组分也可以在拉曼光谱中被分离并鉴定其化学基团。拉曼检测器应使用市售标准蛋白质和蛋白质与小分子、核酸或碳水化合物的复合物进行测试。此外，核-壳纳米颗粒上的稳定剂分子应被研究，因为它们的原位化学组成应提供用于纳米颗粒合成的优化的有价值的信息，这不能通过现有方法适当地获得。