Project Summary The Health Sciences Metabolomics and Lipidomics Core (HSMLC) at the University of Pittsburgh is requesting funds for a Thermo Fisher Exploris 240 mass spectrometer coupled to a Vanquish UHPLC to expand technological capabilities and offerings to its user base. The HSMLC was founded in 2017 and currently serves over 100 investigators at Pitt and beyond. These collaborations range from fee-for-service customers to extensive grant-based collaborations. Since its inception, the core has contributed to 23 peer reviewed publications and 115 grant submissions with a 21% funding rate. The Exploris 240 coupled to the Vanquish UPLC will complement an existing Thermo Fisher ID-X tribrid that was purchased at the end of 2018 with an S10 award. The Vanquish-ID-X system has been used successfully and continuously for both lipidomics and metabolomics since its installation, with its only major downtime attributed to COVID19. Data obtained on the ID-X contributed to 8 of the 23 publications associated with the HSMLC. While many investigators utilize the HSMLC for metabolomics and lipidomics, this proposal highlights investigators whose primary interest is in metabolomics. Installation of the Exploris 240 will complement the ID-X and allow the separation of metabolomic and lipidomics platforms with the Exploris 240 being solely used for metabolomics analysis that includes targeted profiling, differential analysis and 13C tracer studies. Several advancements in instrumentation justify the need for this request over other high resolution mass spectrometers and include (1) Easy IC—internal calibration (2) expanded mass to charge range, down to 40 m/z (3) polarity switching with high mass accuracy and (4) Acquire X functionality for untargeted metabolomics. These advancements will aid in metabolite identification using MS2 fragments in the lower mass range. MS2 fragments between 40-50 m/z will also be critical in the implementation of isotopomers analysis when not only the number of carbons enriched, but the position of enrichment will be required to provide supporting data for metabolic dysregulation. Furthermore, separation of lipidomics and metabolomics onto individual platforms will complement the recent expansion of our core staff and allow for method development time to expand core capabilities and the inception of a more extensive education and training arm of our mission, to provide our expertise in LC-MS analysis, experimental planning and training to collaborators who are interested in better understanding mass spectrometry applications.