This ORIP shared instrumentation proposal requests funding to purchase an Applied Biosystems SCIEX QTRAP 6500(R) triple quadruple / linear ion trap mass spectrometer, a WATERS Acquity(R) ultra high pressure liquid chromatography system, and the BIOCRATES MetIDQ"""""""" metabolite analysis kit. This instrument will be housed in the Biomedical Mass Spectrometry (BioMS) Center at the University of Pittsburgh;a newly created campus-wide center for proteomics and metabolomics. This integrated LC-MS/MS system will be used almost exclusively for the quantification of metabolites from five compound classes that have been identified as members of key biochemical pathways. Furthermore, the unique ion trapping capabilities of the QTRAP trap will be used to support elementary metabolite identification experiments and new assay development. Overall, the acquisition of this instrument will greatly enhance 7 NIH funded projects by enabling nine investigators to optimize well-controlled in-house analyses on valuable samples that are being studied as part of a focused effort to enhance the current understanding of energy metabolism disorders, improve newborn screening, and measure metabolic . Triple quadrupole LC-MS/MS is an essential service for the center, and one five year-old Thermo- Fisher Quantum Ultra is the only instrument of this type in the facility. This sole instrument is configured for proteomic analyses that employ nano-flow separations and tiny capillary columns. As configured, this instrument is not suitable for metabolite assays that use micro-flow separations and narrow bore columns. Furthermore, it is not practical to operate this one system as a """"""""dual-purpose"""""""" instrument that is switched between protein and metabolite analysis without incurring a loss in productivity due to the significant switchover downtime. For these reasons, it has been difficult to meet the current and growing needed for metabolite quantification. Multiple NIH funded investigators have resorted to sending samples out to a patchwork of contract labs that provide limited assay capabilities and variable results. Therefore, the reasons for this application are to: 1) Provide a critically needed in-house metabolite quantitation capability;2) Enable the development and testing of customized metabolite assays that are not easily obtained from contract labs. Relevance The overall impact of this new instrumentation to human is significant, as 9 independent researchers at the University of Pittsburgh (UP) including members of Children's Hospital (CH), and the University of Pittsburgh Cancer Institute (UPCI) require in-house research tools that are needed to improve and advance NIH funded research into the basic biology of energy metabolism, fatty acid oxidation, mitochondrial metabolism, and phenylalanine toxicity. LC-MS/MS based metabolic profiling is not currently available at the University of Pittsburgh. The acquisition of an AB SCIEX QTRAP 6500(R) triple quadruple / linear ion trap mass spectrometer would, for the first time, give these investigators and trainees access to this type of instrumentation, and allow a full complement of metabolite profile data to be under well controlled conditions.
This ORIP shared instrumentation proposal requests funding to purchase an Applied Biosystems SCIEX QTRAP 6500(r) triple quadruple / linear ion trap mass spectrometer, a Waters Acquity(r) ultra high pressure liquid chromatography system, and the BIOCRATES MetIDQ(tm) metabolite analysis package. This instrument will be housed in the Biomedical Mass Spectrometry Center at the University of Pittsburgh;a newly created campus-wide center for proteomics and metabolomics. This integrated LC-MS/MS system will be used almost exclusively for the quantification of metabolites from five compound classes and provide essential data for 12 NIH funded grants awarded to 9 investigators who have significant and growing needs for mass spectrometry-based metabolite measurements. The primary reasons for this application are to: 1) Provide a critically needed in-house metabolite quantitation capability;2) Enable the development and testing of customized metabolite assays that are not easily obtained from contract labs.
| Jackson, Laura E; Kulkarni, Sucheta; Wang, Huabo et al. (2017) Genetic Dissociation of Glycolysis and the TCA Cycle Affects Neither Normal nor Neoplastic Proliferation. Cancer Res 77:5795-5807 |
