7200 accurate mass q tof gc ms

Metabolites from the same treatments described in the above transcriptomic section (2 Gy and 2.5 µM Cur-SLN) were extracted and analyzed by gas chromatography-mass spectrometry (GC-MS), as previously described^{26 (link)}. Derivatization was performed using automated sample prep WorkBench instrument (Agilent Technologies). Dried polar metabolites were dissolved in 60 μl of 2% methoxyamine hydrochloride in pyridine (Pierce) and held at 40 °C for 6 hrs. After dissolution and reaction, 90 μl of MSTFA (N-Methyl-N-(trimethylsilyl) trifluoroacetamid) were added and samples were incubated at 60 °C for 1 h. GC/MS analysis was performed using 7200 accurate-mass Q-TOF GC/MS (Agilent Technologies) equipped with a 40-m DB-5MS capillary column operating under electron impact (EI) ionization at 70 eV. Samples (2 μl) were injected in a splitless mode at 250 °C, using helium as the carrier gas at a flow rate of 1 ml/min. The GC oven temperature was held at 100 °C for 2 min and increased to 325 °C at 10 °C/min.
GC/MS data processing was performed using Agilent Muss Hunter software and statistical analyses were performed using Mass Profile Professional (MPP) software (Musharraf et al., 2016). Relative metabolites abundance was carried out after normalization to internal standard d27-myristic acid and cell number.

Tissue metabolite extraction and derivatization were performed as described in Gaglio et al. [50 (link)]. GC/MS analysis was performed using 7200 accurate-mass Q-TOF GC/MS (Agilent Technologies) equipped with a 40-m DB-35MS capillary column operating under electron impact (EI) ionization at 70eV. Samples (1 μl) were injected in a splitless mode at 250°C, using helium as the carrier gas at a flow rate of 1 ml/min. The GC oven temperature was held at 100°C for 2 min and increased to 325°C at 10°C/min. GC/MS data processing was performed using Agilent Muss Hunter software and statistical analyses were performed using Mass Profiler Professional (MPP) software.