Enhanced fluidity liquid chromatography for biological applications
enhanced fluidity liquid
hydrophilic interaction liquid chromatography
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Publisher:The Ohio State University
Series/Report no.:The Ohio State University. Department of Chemistry Honors Theses; 2010
The following work describes the successful application of enhanced fluidity liquid (EFL) mobile phases to improving isocratic chromatographic separation of biological molecules in hydrophilic interaction liquid chromatography (HILIC) mode. The mobile phase was buffered methanol/water with carbon dioxide added to create the enhanced fluidity liquid. Nucleosides (adenosine, uridine, cytidine, guanosine) were employed as the test sample for this method comparison. Using UV detection at 262 nm, the separation of the sample molecules was studied under each mobile phase condition. Increases in peak resolution between all four were observed as a function of increasing additions of carbon dioxide to create the enhanced fluidity liquid. This increase in resolution was achieved by a combination of improvements in method selectivity and separation efficiency. Plate height was seen to decrease by up to 27%, retention increased for all compounds, and the separation factor for the originally co-eluted cytidine-uridine peak pair increased from 1.03 to 1.24. This resulted in the final baseline resolution of cytidine and uridine. This study marks the first report of EFL being used in conjunction with HILIC.
Ohio State University College of Arts and Sciences Undergraduate Research Scholarship
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