Polar metabolites were extracted and analyzed as described previously [45 (link)], employing a Q Exactive GC Orbitrap GC–tandem mass spectrometer and Trace 1300 Gas chromatograph (Thermo Fisher). Samples were analyzed in three biological replicates.
Q exactive gc orbitrap
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The Q Exactive GC Orbitrap is a high-resolution mass spectrometer designed for gas chromatography-mass spectrometry (GC-MS) applications. It features an Orbitrap mass analyzer that provides accurate mass measurement and high-resolution capabilities for qualitative and quantitative analysis of complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Tracefinder 4, by Thermo Fisher Scientific (2 mentions)
Trace 1300 gas chromatograph, by Thermo Fisher Scientific (1 mentions)
Tg 5silms gc column, by Thermo Fisher Scientific (1 mentions)
Tracetm1300, by Thermo Fisher Scientific (1 mentions)
Mzcloud, by Thermo Fisher Scientific (1 mentions)
Compound discoverer 2, by Thermo Fisher Scientific (1 mentions)
Tg 5silms, by Thermo Fisher Scientific (1 mentions)
Mass frontier 7, by Thermo Fisher Scientific (1 mentions)
Kinetex c18 column, by Phenomenex (1 mentions)
6 protocols using q exactive gc orbitrap
1
Polar Metabolites Analysis by GC-MS
2
GC-MS Analysis of Trimethylsilylated Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The trimethylsilylated samples were injected (2 μL injection volume) into a Q Exactive GC Orbitrap (Thermo, Bremen, Germany). Due to the different derivatization technique, the system was operated with modified chromatographic conditions adapted from routine protocols (Thevis et al., 2011 (link)). The GC was equipped with a HP-Ultra 1 column (17 m × 0.2 mm) of 0.11 mm film thickness. The temperature program started at 180°C and raised with 3°C/min to 240°C and with 40°C/min to 320°C, where it remained constant for 2 min. Samples were injected in split mode with a split flow of 5 mL/min. The mass range for full MS experiments was m/z 50–700 and a resolution of 60,000 FWHM was applied. Data were evaluated with Xcalibur software.
3
GC-MS Metabolite Profiling Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polar metabolites were extracted as previously described with few modifications (Cerna et al., 2017 (link)) and measured using a Q Exactive GC Orbitrap GC-tandem mass spectrometer and Trace 1300 Gas chromatograph (Thermo Fisher). Samples were injected using the split mode (inlet temperature 250°C, splitless time 0.8 min, purge flow 5.0 ml/min, split flow 6.0 ml/min) onto TG-5SILMS GC Column (Thermo Fisher, 30 m × 0.25 mm × 0.25 μm) with helium as a carrier gas at a constant flow of 1.2 ml/min. Metabolites were separated with a 28 min gradient (70°C for 5 min followed by 9°C per min gradient to 320°C and finally 10 min hold time) and ionized using the electron ionization mode (electron energy 70 eV, emission current 50 μA, transfer line and ion source temperature 250°C). The MS operated in the full scan mode, 60000 resolution, scan range 50–750 m/z, automatic maximum allowed injection time with automatic gain control set to 1e6, and lock mass [m/z]: 207.0323. Data were analyzed by TraceFinder 4.1 with Deconvolution Plugin 1.4 (Thermo) and searched against NIST2014, GC-Orbitrap Metabolomics library and inhouse library. Only metabolites fulfilling identification criteria (score ≥ 75 and ΔRI < 2%) were included in the final list.
4
High-Resolution Mass Spectrometry for Compound Identification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HRMS was conducted on a Q-Exactive GC Orbitrap platform coupled with a TraceTM 1300 gas chromatograph and equipped with a programmable temperature vaporizing injector, provided by Thermo Scientific (Bremen, Germany). The GC column was the same as that used for GC-MS/MS analysis. The inlet temperature was set at 90°C. The split flow was 5.0 mL/min with a split ratio at 5:1. The GC gradient was as follows: 45°C for 2 min; heat rate at 50°C/min up to 150°C followed by a rate of 70°C/min until 260°C (hold for 16 min). Total run time was 22 min. Acquisition was achieved in positive electron impact ionisation mode (EI+, 70 eV) with an ion source temperature set at 250°C. The transfer line was maintained at 280°C. Data were recorded in full scan mode (profile) with a resolving power set at 60000 FWHM at m/z 200 and the automatic gain control target defined at 10 6 . The m/z range of the scan was 30-100. Nitrogen gas was used for the C-Trap supply.
5
Multi-Omics Analysis of Organic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GC analyses were carried out on a Thermo Scientific Q Exactive GC Orbitrap. Data were acquired in SIM-SCAN mode from 35-450 AMU. The chromatographic separation was performed on a Thermo Scientific TG-5SILMS (30 m x 250 μm x 0.25 μm) column using Helium carrier as a carrier gas at a constant flow of 1.5 mL/min. Gradient was as follow: initial temperature of 50 °C was held for 5 minutes then ramped to 320 °C at a rate of 15 °C/min and held for 20 minutes, followed by return to 50 °C. Data were analysed with TraceFinder 4.1 and Mass Frontier 7.0 (Thermo Fisher, US) and identification was based on NIST library.
LC analyses were performed on Thermo Scientific Q Exactive LC Orbitrap using full scan mode from 70-1000 AMU. Chromatography was carried out using a Phenomenex Kinetex C18 column (3mm, 100 mm, 2.6 µm). Data were analysed using Mass Frontier 7.0, Compound Discoverer 2.1 (Thermo Fisher) and identification was based on library build with m/z Cloud (Thermo Fisher) and Chemspider.
LC analyses were performed on Thermo Scientific Q Exactive LC Orbitrap using full scan mode from 70-1000 AMU. Chromatography was carried out using a Phenomenex Kinetex C18 column (3mm, 100 mm, 2.6 µm). Data were analysed using Mass Frontier 7.0, Compound Discoverer 2.1 (Thermo Fisher) and identification was based on library build with m/z Cloud (Thermo Fisher) and Chemspider.
6
Volatile Compound Identification by GC-MS/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The volatile compounds were identified by GC-MS/MS (QExactive™ GC Orbitrap™, Thermo-Fisher, Villebon, France), using BP5 MS capillary column (SGE, 30 m x 0.25 mm, 0.25 µm). The carrier gas was helium at a flow rate of 1.2 mL/min. The oven temperature was initially held at 40 °C for 2 min after injection, then increased to 350 °C with a 3 °C/ min heating ramp and kept at 350 °C for 2 min. The PSEO (1 µL) was injected at 250 °C on mode splitless or with split ratio of 1/5. Mass spectrometer monitoring and spectral processing were released by Xcalibur software (Ver. 4.0.27) and the 2014 NIST database and an internally elaborated HRMS library.
Mass spectra of electronic ionization and chemical ionization were recorded. The HRMS system acquired data in the range of 50-550 m/z at 60,000-resolution mode with an energy of 70 eV for electronic ionization. Methane was used at a flow rate of 1.5 mL/min in chemical ionization. The spectrometer was calibrated externally.
Mass spectra of electronic ionization and chemical ionization were recorded. The HRMS system acquired data in the range of 50-550 m/z at 60,000-resolution mode with an energy of 70 eV for electronic ionization. Methane was used at a flow rate of 1.5 mL/min in chemical ionization. The spectrometer was calibrated externally.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!