Untargeted and Targeted Metabolomic Profiling

All samples were analyzed in random order on a Thermo Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer coupled with a Thermo DIONEX UltiMate 3000 HPLC system (Thermo Fisher Scientific, Waltham, MA, USA). For untargeted metabolic profiling, the UltiMate 3000 HPLC system was equipped with a parallel 2DLC (a reversed-phase chromatography (RPC) column and a hydrophilic interaction liquid chromatography (HILIC) column) [22 (link)]. For the targeted metabolomic study, the UltiMate 3000 HPLC system was equipped with an RPC column [21 (link)]. To obtain full MS data, every sample was analyzed by LC-MS in positive and/or negative modes depending on the chemical properties of the metabolites of interest. One pooled sample of each group was analyzed by LC-MS/MS to acquire MS/MS spectra at three collision energies (20, 40, and 60 eV) for metabolite identification.

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He L., Vatsalya V., Ma X., Klinge C.M., Cave M.C., Feng W., McClain C.J, & Zhang X. (2022). Metabolic Analysis of Nucleosides/Bases in the Urine and Serum of Patients with Alcohol-Associated Liver Disease. Metabolites, 12(12), 1187.

Publication 2022

Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer Thermo DIONEX UltiMate 3000 HPLC system

Hplc Hybrid Hydrophilic interaction Liquid chromatography Ms ms Properties of chemical Reversed phase chromatography

Corresponding Organization : Louisville VA Medical Center

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Variable analysis

independent variables

Chromatography column type (reversed-phase chromatography (RPC) column and hydrophilic interaction liquid chromatography (HILIC) column)
Ionization mode (positive and/or negative)

dependent variables

Metabolite profiles (untargeted metabolic profiling)
Targeted metabolite levels

control variables

Instrument (Thermo Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer)
HPLC system (Thermo DIONEX UltiMate 3000)
Collision energies (20, 40, and 60 eV) for MS/MS experiments

controls

Pooled sample of each group analyzed by LC-MS/MS to acquire MS/MS spectra at three collision energies for metabolite identification

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