265b Pressurized Planar Electrochromatography: a New Tool for Proteomics?

David Nurok1, Allyson L. Novotny1, Robert E. Santini2, Randall W. Replogle2, and Gregory L. Hawkins2. (1) Department of Chemistry and Chemical Biology, Indiana University - Purdue University Indianapolis, 402 North Blackford Street, Indianapolis, IN 46202, (2) Chemistry, Purdue University, 560 Oval Drive, West Lafayette, 47907

Pressurized Planar Electrochromatography (PPEC) is a new separation technique that, with respect to both separation speed and chromatographic efficiency, is more similar to High Performance Liquid Chromatography (HPLC) than to classical Thin Layer Chromatography (TLC). The mobile phase is driven by electroosmotic flow, while the sorbent layer is pressurized in such a manner that the separation temperature is controlled. PPEC retains the attractive features of TLC, and this includes the ability to simultaneously separate multiple samples and to perform true two-dimensional separations. For this reason the technique appears to have promise in the field of proteomics.

Our presentation will discuss the apparatus for PPEC, and the dependence of separation quality on variables that include the applied electric field, the applied pressure, the temperature of separation, the buffer concentration and the nature of the sorbent layer. The separation of nine compounds in two minutes illustrates the speed of analysis. Under optimum conditions an extrapolated efficiency of over 100,000 plates per meter is obtained. The presentation will conclude with the discussion of the separation of individual peptides as well as the separation of mixtures of peptides obtained by hydrolysis of proteins.