Research Abstracts Online
2008 - March 2009
University of Minnesota Twin Cities
Institute of Technology
Department of Chemistry
PI: Peter W. Carr
Advantages of High-performance Liquid Chromatography With Ultra-stable Phases
This group’s major goal is to increase the speed and effectiveness of prior work in developing high-speed, comprehensive two-dimensional liquid chromatography (2DLC) based on high-temperature, ultra-fast (21 sec. cycle time) gradient elution chromatography. To date, their fast 2DLC systems can produce peak capacities > 1000 in less than 30 minutes. To expand on these accomplishments, they will develop specifically designed, highly stable "hyper-crosslinked” stationary phases for use as the ultra-fast second dimension of 2DLC. They have initiated the development of dual reversed-phase, dual gradient 2DLC (2DLC which uses gradient elution and reverse phase liquid chromatography in both dimensions). They believe that the dual reverse-phase approach is the most powerful for a wide variety of biological mixtures. Major advances can be made through: the study of the fundamental processes controlling the speed and effective peak capacity of 2DLC; by improving the instrumentation needed to do 2DLC especially the "dynamic gradient elution” concept; by providing additional novel, ultra-stable stationary phases offering alternative selectivities and tuned retentivity; and by developing data analysis methods to deal with the insidious retention time (temporal) alignment problem and implement multi-variate analysis of the huge data sets (>100 Mbytes) collected during a single high-speed two-dimensional run with a multi-wavelength detector.
Studies obtained on real samples strongly suggest that the separating power of 2DLC becomes superior to that of one-dimensional LC in only 5-10 minutes. If this proves generally true, 2DLC becomes interesting not only for very complex samples but also for more typical analytical problems. If the speed of the second dimension can be increased then this will be so. The researchers’ chief interest is in developing and applying these methods to problems in plant, pharmaceutical, environmental, and medical metabolomics, which they believe is a bio-analytically very important challenge in many ways quite different from proteomics.
Dwight R. Stoll, Department of Chemistry, Gustavus Adolphus College, Saint Peter, Minnesota