Targeted Metabolite Profiling with LC-MS

Metabolite extracts (10 μl) were loaded onto a Luna-amino column (Phenomenex) using the UltiMate 3000 TPLRS LC device as previously described (Xiao et al., 2020 (link)). Subsequent analysis was performed using a Q-Exactive HF-X mass spectrometer (Thermo Fisher Scientific) as previously described (Xiao et al., 2020 (link)). Targeted processing of a subset of known metabolites was conducted using TraceFinder software version 4.1 (Thermo Fisher Scientific). Compound identities were confirmed using reference standards. In all cases, metabolite abundance was normalized using internal standards, and relative changes were assessed by comparison with metabolite extracted from the same sample type (i.e., plasma). Metabolomics data were analyzed and principal component analysis plots were generated using MetaboAnalyst 5.0 software.

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Dyck L., Prendeville H., Raverdeau M., Wilk M.M., Loftus R.M., Douglas A., McCormack J., Moran B., Wilkinson M., Mills E.L., Doughty M., Fabre A., Heneghan H., LeRoux C., Hogan A., Chouchani E.T., O’Shea D., Brennan D, & Lynch L. (2022). Suppressive effects of the obese tumor microenvironment on CD8 T cell infiltration and effector function. The Journal of Experimental Medicine, 219(3), e20210042.

Publication 2022

Luna-amino column Q-Exactive HF-X mass spectrometer TraceFinder software version 4.1

Device Plasma

Corresponding Organization : Brigham and Women's Hospital

Other organizations : University College Dublin, St. Vincent's University Hospital, Mater Misericordiae University Hospital, Dana-Farber Cancer Institute, National University of Ireland, Maynooth, National Children’s Research Centre

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Variable analysis

independent variables

Metabolite extracts (10 μl)

dependent variables

Metabolite abundance

control variables

Metabolite extracted from the same sample type (i.e., plasma)
Internal standards

controls

Reference standards used to confirm compound identities

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