(741a) Novel Approach to Employing SFC for Metabolomics Research

Metabolites, the end products of cellular regulatory processes, can be regarded as the ultimate response of biological systems to genetic, pathophysiological or environmental stressors. Human serum target analysis, metabolic profiling, metabolic fingerprinting, or broadband metabolomic analysis can therefore reflect the pathological state of various organs and aid in the early detection of disease. Whereas proteomic and genomic technologies are more mature, metabolome analysis tools, which rely heavily on chemical separations, are still constantly evolving.

A substantial challenge is the need to identify and analyze accurately potential metabolic biomarkers. Supercritical Fluid Chromatography (SFC) has the capability to separate rapidly a mixture of small molecules based on their specific volatility in supercritical fluids. More important, however, are the added advantages of higher efficiency and better resolution at ambient conditions than traditional HPLC and/or GC. SFC also significantly reduces the amount of liquid solvent, minimizing potential undesirable interferences with the subsequent mass spectrometry analysis. The limited solubility of many common metabolites in the supercritical fluid phase (usually scCO2), however, is an obstacle keeping researchers from using SFC for metabolomics.

The project thus has two main thrusts. We have first focused on enhancing metabolite solubility by chemical modification using a silylation reagent to derivatize amino acids and small peptides. Eventually this will lead to derivatizing serum metabolites, with the ultimate goal of targeting complete metabolomes in physiological samples. The second part of the project involves the SFC separation and subsequent analysis via UV of model metabolites.