AMINO ACID ANALYSIS USING MICELLAR ELECTROKINETIC CAPILLARY CHROMATOGRAPHY

使用胶束电动毛细管色谱法进行氨基酸分析

• 批准号: 6107002
• 负责人: JOEL T SMITH
• 金额: $ 7.69万
• 依托单位: SOUTHEASTERN OKLAHOMA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6107002
• 关键词: aminoacid; aminoacid analyzer; capillary electrophoresis; method development; trace elements

The trace analysis of biological components has been a critical key towards the development of modern biochemistry and medicine. One particularly powerful contribution has been the development of separations techniques capable of performing rapid and quantitative analysis on small amounts of biogenic materials. High performance liquid chromatography (HPLC) has proven capable of resolving all naturally occurring amino acids in a relatively short time frame providing a robust method for the determination of amino acid composition of proteins and peptides. One significant drawback to HPLC, as well as many other currently available analysis methods, is the quantity of biological sample required to perform the analysis since heroic efforts are often required to isolate the components(s) of interest. As a conseuence, more sensitive microanalysis separation schemes are being developed to provide increased mass sensitivities and improved resolving capabilities. Among these new techniques, capillary electrophoresis (CE) has proved to be an exceptionally capable and cost effective tool. The overall objective of this project is to extend the utility of micellar electrokinetic capillary chromatography (MECC), a mode of CE, towards the microanalysis of biological materials. The proposed studies will evaluate a novel series of organic additives used to promote changes in the electrochromatographic selectivity towards biogenic molecules in micellar systems at low concentrations. More specifically, the aim is to provide a rapid and reproducible separation scheme that produces complete resolution of all naturally occurring amino-acids with femto- mole mass sensitivities. This objective will be approached by evaluating a novel series of organic additives combined with traditional micellar phases to determine their effects on electrochromatographic retention and selectivity through a series of systematic studies altering each chromatographic parameter independently. Upon a firm understanding of the behavior of these additives in micellar systems, optimization of a MECC method for the quantitative determination of derivatized amino acids will be addressed. This project will provide a better understanding of the utility of modified micellar phases in MECC towards biogenic species and if successful provide a cost effective alternative to microanalysis of amino acids.

生物成分的痕量分析已成为关键 促进现代生物化学和医学的发展。 一 特别有力的贡献是开发 能够进行快速定量的分离技术 少量生物材料的分析。 高性能液体 色谱法 (HPLC) 已被证明能够解决所有自然问题 在相对较短的时间内出现的氨基酸提供了强大的 蛋白质氨基酸组成的测定方法 肽。 HPLC 的一个显着缺点以及许多其他缺点 目前可用的分析方法，是生物量 执行分析所需的样本，因为英雄的努力往往是 需要隔离感兴趣的组件。 结果，更多 正在开发灵敏的微量分析分离方案，以提供 提高质量灵敏度并提高分辨能力。 之中 这些新技术，毛细管电泳（CE）已被证明是一种 功能异常且具有成本效益的工具。 该项目的总体目标是扩大 胶束电动毛细管色谱 (MECC)，CE 模式， 致力于生物材料的微量分析。 拟议的研究 将评估一系列用于促进变革的新型有机添加剂 电色谱对生物分子的选择性 低浓度的胶束系统。 更具体地说，目标是 提供快速且可重复的分离方案，产生 用 femto- 完全解析所有天然存在的氨基酸 摩尔质量敏感性。 这一目标将通过评估来实现 与传统胶束相结合的一系列新型有机添加剂 确定它们对电色谱保留的影响 通过一系列系统研究的选择性，改变每一个 独立的色谱参数。 在深刻理解后 这些添加剂在胶束系统中的行为，优化 MECC 方法定量测定衍生氨基酸 将得到解决。 该项目将让人们更好地了解 MECC 中改性胶束相对生物物种的效用 如果成功的话，可以提供一种具有成本效益的微量分析替代方案 氨基酸。