GC-MS Analysis of Non-Polar Metabolites

Non-polar metabolites were derivatized with 295 μL tert-methyl-Butyl-Ether (MTBE) and 5 μL of trimethylsulfonium hydroxide (TMSH) for 30 min at room temperature. The tubes were centrifuged for 3 min at 20,000 g to remove insoluble particles before transferring the supernatants to GC-microvials. GC-MS measurements were carried out following a previously developed procedure (Valledor et al., 2014b (link)) on a triple quad instrument (TSQ Quantum GC; Thermo, United States). The mass spectrometer was operated in electron-impact (EI) mode at 70 eV in a scan range of m/z 40–600. Metabolites were identified based on their mass spectral characteristics and GC retention times through comparison with the retention times of reference compounds in an in-house reference library and the current version of Golm Metabolome Database (Hummel et al., 2007 (link)) using LC-Quant software (Supplementary Table S1b).

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Escandón M., Meijón M., Valledor L., Pascual J., Pinto G, & Cañal M.J. (2018). Metabolome Integrated Analysis of High-Temperature Response in Pinus radiata. Frontiers in Plant Science, 9, 485.

Publication 2018

TSQ Quantum GC

Electron Gc ms Library Metabolome Methyl tert butyl ether Retention Scan Trimethylsulfonium hydroxide

Corresponding Organization : Universidad de Oviedo

Other organizations : University of Turku, University of Aveiro

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Variable analysis

independent variables

Derivatization of non-polar metabolites with 295 μL tert-methyl-Butyl-Ether (MTBE) and 5 μL of trimethylsulfonium hydroxide (TMSH) for 30 min at room temperature

dependent variables

Metabolite identification based on mass spectral characteristics and GC retention times

control variables

Centrifugation of the samples for 3 min at 20,000 g to remove insoluble particles before transferring the supernatants to GC-microvials
GC-MS measurements carried out following a previously developed procedure (Valledor et al., 2014b)
Mass spectrometer operated in electron-impact (EI) mode at 70 eV in a scan range of m/z 40–600

controls

Comparison of metabolite retention times with reference compounds in an in-house reference library and the Golm Metabolome Database (Hummel et al., 2007)

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