Frozen left ventricular tissues from the young-WT-sham, aged-WT-sham, Sesn2-KO-sham, young-WT-I/R, aged-WT-I/R, Sesn2-KO-I/R mice were homogenized to extract polar metabolites for 2DLC-MS/MS analysis with Thermo Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer coupled with a Thermo DIONEX UltiMate 3000 HPLC system (Thermo Fisher Scientific, Waltham, MA, USA) [22 (link)]. Each sample was respectively analyzed by 2DLC-MS in positive mode (+) and negative mode (−) to obtain the full MS data. For 2DLC-MS data analysis, XCMS software was used for spectrum deconvolution, and MetSign software was used for metabolite identification, cross-sample peak list alignment, normalization, and statistical analysis [[23] (link), [24] (link), [25] (link)]. Enrichment analysis of significantly altered metabolites were performed with MBROLE 2.0 by searching against KEGG, UniPathway, and BioCyc SMPDB database to obtain significantly regulated pathways [26 (link)].
Thermo q exactive hf hybrid quadrupole orbitrap mass spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Thermo Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer is a high-performance mass spectrometry instrument that combines a quadrupole mass filter and an Orbitrap mass analyzer. It provides high-resolution, accurate-mass measurements for a wide range of analytical applications.
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Lab products found in correlation
2 protocols using thermo q exactive hf hybrid quadrupole orbitrap mass spectrometer
1
Metabolomic Analysis of Cardiac Ischemia-Reperfusion
2
Comprehensive 2DLC-MS/MS Metabolomics
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2DLC‐MS/MS analysis was completed by first collecting left ventricular tissue samples at 4°C to retain metabolites. Polar metabolites were mixed with methanol and centrifuged to acquire a metabolite supernatant. The supernatant was analyzed by the Thermo Q Exactive HF Hybrid Quadrupole‐Orbitrap Mass Spectrometer coupled to the Thermo DIONEX UltiMate 3000 HPLC system (Thermo‐Fisher Scientific). Utilizing reverse‐phase chromatography (RPC) and hydrophilic interaction chromatography (HILIC) in the HPLC, each sample was analyzed by 2DLC‐MS. Each sample group were split into six pooled samples and analyzed in both positive and negative modes to obtain the full MS/MS spectra at 20, 40, and 60 eV collision energies.
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