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Investigation of the separation mechanism in hydrophilic interaction chromatography.

亲水相互作用色谱分离机理的研究。

基本信息

批准号：
EP/J016578/1
负责人：
David McCalley
金额：
$ 19.36万
依托单位：
University of the West of England
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2012
资助国家：
英国
起止时间：
2012 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FJ016578%2F1
关键词：
Investigation separation mechanism hydrophilic interaction

项目摘要

Chromatography is a method of separating and analysing important chemicals such as ethical pharmaceuticals, drugs of abuse, food additives, pesticides and compounds in biological fluids that may be indicative of disease. A popular variant is high performance liquid chromatography, where the sample is introduced into a liquid stream (the "mobile phase") and passed through a column containing particles of the separation medium (the "stationary phase"). If these particles are made smaller, then the separations improve and they can be accomplished in minutes or even seconds, rather than hours or days. Small particles however, require high pressures to force the liquid through, hence the term high performance (or high pressure) liquid chromatography (HPLC). A detector situated at the end of the column senses the separated chemicals and allows them to be quantified. The annual world market for instruments and consumables in HPLC is several billion dollars; tens of thousands of analyses are carried out daily in the UK alone. Thus, faster and more efficient variations of the technique are continually being sought. One of these is hydrophilic interaction chromatography (HILIC), a technique that has significant advantages over the commonly used separation procedure (reversed-phase, RP), particularly for the fast analysis of drugs and compounds of clinical significance. Many of these compounds have only weak interactions with the usual RP columns and thus give poor separations. However, the mechanism of HILIC is poorly understood, which makes it difficult to design the separation conditions. Some separations seem to work very well, whereas others do not, for no apparent reason. The aim of this research is to elucidate the principles of the HILIC separation mechanism and to elucidate the experimental parameters that are most influential on the separation. This knowledge will be used to develop a "tool box" approach to apply the technique to separations that have proved difficult by conventional techniques. The idea is to apply an automated series of HILIC separations using different experimental conditions that generate different selectivity, such that an optimum for the analysis can be chosen. The study is of particular importance to the pharmaceutical industry especially in connection with new methods of drug design, such as "fragment based drug discovery" which involve synthesis of more complex structures using small building blocks each containing some desirable property of the final compound. Many of these building blocks are small polar molecules that should be very well suited to the HILIC technique. All pharmaceuticals need to be carefully screened for the presence of impurities, that may give rise to undesirable physiological side effects. HILIC can be used for example to assess the presence of residues of these polar starting materials in the final product. HILIC appears less susceptible to "overload", a phenomenon where distortion of the separation takes place when larger amounts of material are analysed. Overload can cause interference between large peaks of the active pharmaceutical ingredient and small quantities of impurities.The proposed work will require the development of a mixture of compounds of different structure that can be used to probe the various contributions to retention in HILIC. While the HILIC technique is very simple in practice, it appears that there are many complex factors which contribute to the separations. For example, many important pharmaceuticals, drugs of abuse, and compounds of biomedical significance are charged species, which adds an extra dimension of complexity to the separation mechanism. Understanding the importance of these various contributions will allow separations to be designed which can fully exploit the useful differences between HILIC and conventional separation methods.

层析是一种分离和分析重要化学物质的方法，如道德药物、滥用药物、食品添加剂、杀虫剂和生物液中可能表明疾病的化合物。一种流行的变种是高效液相色谱，将样品引入液体流(“流动相”)，并通过含有分离介质颗粒的柱(“固定相”)。如果这些颗粒变得更小，那么分离效果就会改善，它们可以在几分钟甚至几秒内完成，而不是几小时或几天。然而，小颗粒需要高压才能迫使液体通过，因此术语高性能(或高压)液相色谱(HPLC)。位于色谱柱末端的检测器可以检测分离出的化学物质，并对它们进行定量。高效液相色谱仪器和耗材的年度全球市场价值数十亿美元；仅在英国，每天就进行数万次分析。因此，人们一直在寻求更快、更有效的技术变体。其中之一是亲水相互作用层析(HILIC)，这项技术比常用的分离程序(反相，RP)具有显著优势，特别是在快速分析具有临床意义的药物和化合物方面。其中许多化合物与通常的反相色谱柱只有很弱的相互作用，因此分离效果很差。然而，目前对HILIC的分离机理还不是很清楚，这给分离条件的设计带来了困难。有些分手似乎效果很好，而另一些则不是，没有明显的原因。本研究的目的是阐明HILIC分离机理，并阐明对分离影响最大的实验参数。这些知识将被用来开发一种“工具箱”方法，将该技术应用于传统技术证明困难的分离。这个想法是应用一系列自动化的HILIC分离，使用不同的实验条件产生不同的选择性，这样就可以选择分析的最佳条件。这项研究对制药业特别重要，特别是在药物设计的新方法方面，例如“基于片段的药物发现”，它涉及用小的积木合成更复杂的结构，每个积木包含最终化合物的一些理想性质。这些构件中的许多都是小的极性分子，应该非常适合HILIC技术。所有药物都需要仔细检查是否存在杂质，这些杂质可能会引起不良的生理副作用。例如，HILIC可用于评估这些极性起始物质在最终产品中的残留量。HILIC似乎不太容易受到“过载”的影响，这是一种现象，当分析更多的材料时，分离会发生扭曲。过载会导致有效药物成分的大峰和少量杂质之间的干扰。拟议的工作将需要开发不同结构的化合物的混合物，可以用来探索在HILIC中保留的各种贡献。虽然HILIC技术在实践中非常简单，但似乎有许多复杂的因素导致了分离。例如，许多重要的药物、滥用药物和具有生物医学意义的化合物都是带电物种，这为分离机制增加了额外的复杂性。了解这些不同贡献的重要性将有助于设计能够充分利用HILIC和传统分离方法之间的有益差异的分离。