Using Mass Spectrometry to Quantitate and Analyze Bioactive Small Molecules

Metabolomics is the study of small molecules in a biological system. Metabolomics encompasses a wide range of research questions, and therefore requires the use of varied workflows. This thesis contains introduction to targeted and untargeted metabolomics, kinds of mass spectrometers used for each, as well as what kinds of experiments fall under each category. The first project applies fundamental targeted metabolomics techniques to a multiple reaction monitoring assay of catecholamines in cell media to investigate the role of catecholamines as chemical signals in the primary metastasis of epithelial high grade serous ovarian cancer. The majority of the work done focused on optimizing sample preparation, involving the extraction of catecholamines of a DMEM agarose well and matrix removal. The second project details the process of cultivating a unique mammalian-sourced microbial library through untargeted metabolomics coupled with bioinformatics tools.The project focuses on the initial steps involved with developing a microbial library for drug discovery, starting with environmental isolation. This resulted in selecting 37 strains for a drug discovery library from 273 bacterial and 50 fungal isolates through pseudotaxonomic diversity and metabolite diversity.