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.

该项目旨在建立一种用于非对称流场流分级（AF4）的在线拉曼检测系统。 AF4 是一种大分子分离方法，还可以确定扩散系数的分布。流体动力学尺寸范围为 1 nm 至 100 微米。当前的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成分的紫外-可见光谱或荧光光谱。所提出的拉曼检测引入了获得有关样品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，因为使用紫外-可见光和荧光检测器区分许多样品是必要的。与我们的建议相当的方法只有在巨大的财务费用和严格限制可用洗脱溶剂数量的情况下才可用。即使是完全未知的样品的单一成分也可以被分离，并且它们的化学基团可以在拉曼光谱中被识别。拉曼检测器应使用市售的标准蛋白质和小分子、核酸或碳水化合物的蛋白质复合物进行测试。此外，还应研究核-壳纳米颗粒上的稳定剂分子，因为它们的原位化学成分应该为纳米颗粒合成的优化提供有价值的信息，而现有方法无法正确获得这些信息。