Method Development for Measuring Central Metabolites

Mentors: Clary Clish and Amanda Souza

Metabolite profiling is the systematic analysis of metabolites to study metabolism, and elucidate metabolic signatures of physiologic cellular processes, in both healthy and diseased cells. Metabolites in biological samples are quantified through the liquid chromatography tandem mass spectrometry (LC-MS) technology. Comparative analyses of data obtained from LC-MS can then be used to associate levels of certain metabolites with disease states.

Sarah’s project goal was to address some of the known limitations of two current metabolite profiling LC-MS methods: ion pairing chromatography (IPC) and hydrophilic interaction chromatography (HILIC). Sarah tested combinations of three different chromatography types and 5 different mobile phase conditions, by profiling metabolites from reference samples, cell extract samples, and pooled plasma samples. Data was then assessed and scored for peak quality. Sarah found a modification in IPC that yielded better peak quality scores and coefficients of variation. In HILIC, Sarah found that using a different column for chromatography allowed for more complete resolution and greater stability in some samples; however, the original column produced better peak qualities and coefficients of variation in other samples. 



Sarah, a senior at Malden High School, found ways to improve to the LC-MS technologies that are used to analyze the levels of small metabolites in cell extract and pooled plasma samples.