Gas chromatography–mass spectrometry (GC–MS) was done by a TRACE GC Ultra Gas Chromatograph coupled to an ITQ 900 GC-Ion Trap MS (both Thermo ScientificTM). The instrument was equipped with a Rtx® -5MS column (30 m, iD 0.25, df 0.25 μm; Restek). Helium was used as carrier gas with a constant flow of 1 ml/min. 1 μl sample was injected (split less) for GC–MS analysis. The injector temperature was set to 250°C using a Trance & Focus GC-Liner. Before and after each injection the syringe was washed 10 times with 10 μl chloroform and hexane each. The oven program was: 3 min 80°C, ramp with 5°C per min up to 325°C, 2 min 325°C. The transfer line temperature was set at 250°C and the ion source at 220°C. Mass spectra were recorded at two scans per s from m/z 50 to 750. Electron energy was set to 70 eV. Raw data were converted to cdf-files by Xcalibur 2.0.7 software (Thermo) and uploaded to MeltDB (Neuweger et al., 2008 (link)). Ribitol was found to be absent in the samples and it was therefore possible to add it as an internal standard.
Itq 900 gc ion trap ms
Manufactured by Thermo Fisher Scientific
The ITQ 900 GC-Ion Trap MS is a gas chromatography-ion trap mass spectrometer (GC-Ion Trap MS) system manufactured by Thermo Fisher Scientific. The core function of this instrument is to separate, detect, and analyze complex chemical mixtures using gas chromatography coupled with ion trap mass spectrometry technology.
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2 protocols using itq 900 gc ion trap ms
1
GC-MS Analysis of Metabolite Samples
2
GC-MS Analysis of Metabolites
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GC-MS was done for high abundant compounds by a TRACE GC Ultra gas chromatograph coupled to an ITQ 900 GC-Ion Trap MS (both Thermo ScientificTM). The instrument was equipped with a Rtx ® -5MS column (30 m, iD 0.25, df 0.25 μm; Restek). One μl sample was injected (splitless) for GC-MS analysis. The oven program was: 3 min at 80 °C, ramp with 5 °C/min up to 325 °C, 2 min at 325 °C. Transfer line temperature was set to 250 °C and ion source on 220 °C. Mass spectra were recorded from m/z = 50 to 750. Replicate samples were derivatized and measured separately with intervals of at least three days. A blank to check carry over of metabolites was run every five to six samples. To evaluate constant chromatographic performance and sensitivity, a defined complex biological sample derived from germinating Medicago truncatula seeds was measured once a week. Samples were measured at least in technical triplicates on one biological replicate.
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