IMPROVED USE OF REVERSED-PHASE HPLC COLUMNS

改进反相 HPLC 色谱柱的使用

• 批准号: 2869898
• 负责人: LLOYD R SNYDER
• 金额: $ 10.71万
• 依托单位: LC RESOURCES, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2869898
• 关键词: analytical method; computer program /software; computer simulation; computer system design /evaluation; evaluation /testing; high performance liquid chromatography; mathematical model; physical property; technology /technique development

Alkyl-silica columns (e.g., C8, C18) are presently the most popular by far for use in analyzing samples by means of high-performance liquid chromatography (HPLC). Unfortunately, column manufacturers are currently unable to guarantee that different production batches of nominally equivalent column packing will provide the same separation of different samples, while user surveys indicate that column irreproducibility is a serious problem that can result in major delays and added costs in routine HPLC analysis. The present proposal is intended to solve this problem in two ways. First, a review and extension of previously published studies suggests that it should be possible to characterize different columns in terms of the fundamental column properties that determine sample retention and separation, by the use of a suitable standard sample (mixture of test compounds) and test procedure. It may also be possible to modify these column properties by suitable adjustments in other separation conditions (mobile phase, temperature, flow rate), so as to reduce (and hopefully eliminate) minor differences in column retention for any sample. Second, it is proposed to develop computer software that will allow the user to conveniently predict separation as a function of small, simultaneous variations in any number of experimental conditions (e.g., mobile phase pH, column temperature, etc.) The software would also automatically correct for column-to-column differences in retention by recommending suitable adjustments in separation conditions. This would minimize the impact of column irreproducibility, as well as allow unattended correction for any change in the column during use. This capability is currently needed for high-throughput applications of HPLC, as are found in combinatorial synthesis. Only uses of this new software are described in the proposal. PROPOSED COMMERCIAL APPLICATION: The proposed software and accompanying protocols can be used by (a) column manufacturers to better control column manufacture and testing and (b) chromatographers to develop HPLC methods, to troubleshoot assay procedures and to adapt procedures for column irreproducibility.

烷基-二氧化硅柱（例如，C8，C18）是目前最流行的用于通过高效液相色谱（HPLC）分析样品的方法。不幸的是，色谱柱制造商目前无法保证标称等效色谱柱填料的不同生产批次将提供不同样品的相同分离，而用户调查表明，色谱柱的不可重复性是一个严重的问题，可能导致常规HPLC分析的重大延迟和成本增加。本提案旨在从两个方面解决这一问题。首先，回顾和扩展以前发表的研究表明，它应该是可能的，以确定样品保留和分离的基本列属性，通过使用合适的标准样品（测试化合物的混合物）和测试程序来表征不同的列。也可以通过适当调整其他分离条件（移动的相、温度、流速）来改变这些色谱柱特性，从而减少（并有望消除）任何样品的色谱柱保留的微小差异。第二，建议开发计算机软件，该软件将允许用户方便地预测分离作为任何数量的实验条件（例如，移动的相pH、柱温等）该软件还将通过建议适当调整分离条件来自动校正柱与柱之间的保留差异。这将最大限度地减少色谱柱不重现性的影响，并允许在使用过程中对色谱柱的任何变化进行无人值守的校正。这种能力是目前所需要的高效液相色谱的高通量应用，如在组合合成中发现。提案中仅描述了这种新软件的用途。拟定商业应用：拟议的软件和随附方案可供（a）色谱柱制造商用于更好地控制色谱柱制造和检测，以及（B）色谱分析师用于开发HPLC方法、对测定程序进行故障排除以及调整色谱柱不重现性程序。