Project Summary The requested instrumentation in this proposal is an ultra-high resolution mass spectrometer coupled to an ultra-high performance liquid chromatography system that will be used for lipidomics, metabolomics, isotope tracing, and structural elucidation experiments. Specifically, we are requesting funds for a Thermo Scientific™ Orbitrap™ IQ-X™ Tribrid™ Mass Spectrometer coupled to a Vanquish dual column liquid chromatography system, to expand our technological capabilities and offerings to its user base. This instrument will be housed in the Translational Biomarker Core (TBC) in the Center of Excellence in Toxicology (CEET) at the Perelman School of Medicine of the University of Pennsylvania (Penn). The TBC currently serves over 80 investigators from Penn and beyond. These collaborations range from fee-for-service customers to extensive grant-based collaborations. Until 2016, the TBC only offered targeted quantification assays and proteomics methodologies. In 2016, the Core acquired a Dionex™ Ultimate™ HPG-3400RS ultra high-pressure liquid- chromatography (UPLC) that was interfaced with an Orbitrap QE-HF that was running proteomics using a nano- flow-LC in the Blair laboratory. With limited instrument time, the Core developed its lipidomic platform by combining the HRMS raw data with Lipids Search (Thermo) software for lipids identification. This assay is one of the most requested assays offered by the Core, and through collaborations, we have now more than 300 lipids standards used for calibration curves. During the University restrictions due to Covid-19 in spring 2020, we ran the 600 metabolomics standards commercially available, building a library for Compound Discoverer 3.2 (Thermo). The metabolomics workflow was used for several successful grant submissions during the last two years. The Core would like to expand its capabilities to run these types of highly multiplexed and untargeted omics routinely, to expand technological capabilities, and fit offerings to its user base needs. This proposal highlights the need of omics assays from 29 users (28 with NIH funding). Additionally, the core has established ongoing collaborations with institutes and centers at Penn including Children’s Hospital of Philadelphia (CHOP), the Institute for Translational Medicine and Therapeutics, and the Institute of Immunology. Given the focus of the users on the identification of novel small molecule biomarkers of inflammation and related chronic diseases such as cancer and diabetes, this mass spectrometer is urgent and vital for our research projects. Expertise in the Core includes staff that is responsible for instrument maintenance, sample preparation, method development, and data analysis, including large data sets that require the use of bioinformatics software for differential analysis. Furthermore, having a dedicated HRMS instrument will complement the recent expansion of our Core staff. It will allow method development time to expand core ...