MONOLITIC C18 HPLC AND HILIC/(+)ESI-MS METABOLOMIC PROFILING OF PLASMA SAMPLES

SOLEDAD CERUTTI, JODIE V. JOHNSON, PEGGY R. BORUM, RICHARD A. YOST, DAVID H. POWELL

Tampa, Florida, Estados Unidos

Simposio; 2nd Moffitt Symposium on Mass Spectrometry; 2009

MOFFITT Cancer Center

Small molecule profiling, i.e. Metabolomics, uses mass spectrometry to detect, quantify, and identify the small molecules that are participants in general metabolic reactions and that are required for the maintenance, growth and normal function of cells. The metabolome more closely reflects the activities of cells at a functional level than the genome or proteome as metabolite fluxes are a product of both genetic and environmental influences. Thus, changes in the metabolome are expected to provide insight into the nutritional, metabolic, and general health status during different stages of the life cycle and during pathological conditions.We are testing the hypotheses that the plasma metabolome will change in piglets between days 2 through 8 of life. The putative biomarkers were determined by the analysis of plasma via both monolithic C18 reverse phase chromatography and hydrophilic interaction chromatography (HILIC) approaches coupled to a high resolution time-of-flight mass spectrometer operated in the positive electrospray ionization [(+)ESI] mode. Performance/contribution of each separation strategy, identification of unique and changing m/z features by processing the raw data with the MassHunter and GeneSpring-MS software platforms are presented. From our findings, we are suggesting a general protocol to obtain a reliable and more global view of the metabolome.