Primary compounds were extracted from 100 mg of fresh leaves and then subjected to derivatization using methoxamine hydrochloride in pyridine to inhibit the cyclization of reducing sugars and then with N-methyl-N-trimethylsilyl-trifluoroacetamide with 1% trimethylchlorosilane for trimethylsilylation following the Chitarrini et al. (2017 (link)) procedure. The derivatized extract was then injected for GC/MS analysis using a Trace GC Ultra with a fused silica RXI-5-Sil MS w/Integra Guard (30 m × 0.25 mm × 0.25 μm) column, combined with mass spectrometer TSQ Quantum GC (Thermo Electron Corporation) following the Chitarrini et al. (2017 (link)) parameters.
Tsq quantum gc
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TSQ Quantum GC is a triple quadrupole gas chromatography-mass spectrometry (GC-MS/MS) system designed for high-sensitivity and high-selectivity analysis of complex samples. It delivers accurate, reliable, and reproducible results for a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Unity inova 500 ft nmr, by Agilent Technologies (3 mentions)
Trace gc ultra, by Thermo Fisher Scientific (2 mentions)
Trace gc, by Thermo Fisher Scientific (1 mentions)
Rtx200 column, by Restek (1 mentions)
Spectrum one fourier transform, by PerkinElmer (1 mentions)
Uv 2600 uv vis spectrophotometer, by Shimadzu (1 mentions)
Spectrum one ftir spectrometer, by PerkinElmer (1 mentions)
Hp 5ms capillary column, by Agilent Technologies (1 mentions)
Ultrasonic cleaner, by Emerson (1 mentions)
Balance 0.01 g, by Sartorius (1 mentions)
Factorfour capillary column vf 5 ms, by Agilent Technologies (1 mentions)
11 protocols using tsq quantum gc
1
GC/MS Analysis of Leaf Metabolites
2
GC-MS Analysis of Organic Compounds
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Analyses were performed using a Trace GC Ultra combined with a mass spectrometer TSQ Quantum GC and an autosampler Triplus (Thermo Electron Corporation, Waltham, MA, United States). A RXI-5-Sil MS w/Integra-Guard®(fused silica) (30 m × 0.25 mm × 0.25 μm) column was used for compound separation. Helium was used as the carrier gas at 1.2 mL/min and the injector split ratio was set to 1:10. The injector, transfer line and source temperature were set to 250°C. The initial oven temperature was kept at 65°C for 2 min, increased by 5.2°C/min to 270°C and held at 270°C for 4 min. These conditions were shown to represent a good compromise in order to obtain a not excessively long chromatographic run, a high number of compounds and good peak separation. The mass spectrometer was operated in electron ionization mode. Data acquisition was performed in full scan mode from 50 to 700 m/z. Data processing was performed using XCALIBURTM 2.2 SOFTWARE.
3
GC-MS Analysis of Non-Polar Metabolites
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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 ).
4
Analysis of FTOHs and FTACs
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The analysis of FTOHs and FTACs
was performed on a Trace GC coupled to a TSQ Quantum GC (both Thermo
Fisher) with a RTX200 column (30 m × 0.25 mm × 1.0 μm)
from Restek (Bellefonte, U.S.A.). Ionization was conducted with methane
as a reagent gas. The quantification of FTOHs was performed with the
corresponding internal standard while M10:2 FTOH was used for the
quantification of 6:2 FTAC.
was performed on a Trace GC coupled to a TSQ Quantum GC (both Thermo
Fisher) with a RTX200 column (30 m × 0.25 mm × 1.0 μm)
from Restek (Bellefonte, U.S.A.). Ionization was conducted with methane
as a reagent gas. The quantification of FTOHs was performed with the
corresponding internal standard while M10:2 FTOH was used for the
quantification of 6:2 FTAC.
5
Characterization of Novel Compounds
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All of the reagents and solvents were purchased
as reagent grade and used without further purification. Nuclear magnetic
resonance (NMR) spectra were recorded on a Varian Unity INOVA 500
FT-NMR. Chemical shifts for the samples were measured in DMSO-d6 and calibrated against the sodium salt of
3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP) as an external reference in a sealed capillary tube.
NMR data were processed and analyzed with MestReNova version 6.1.1-6384.
The IR spectra were recorded on a Perkin Elmer-Spectrum One Fourier
transform infrared spectrometer with KBr disks in the range of 4000–400
cm–1. The melting point was determined on a Mel-Temp
(Electrothermal 120 VAC 50/60 Hz) melting point apparatus and was
uncorrected. Mass spectral data were obtained at electrospray ionization
mass spectrometry (ESI-MS) positive mode on a TSQ Quantum GC (Thermo
Scientific). Elemental analysis was carried out using an ECS 4010
Analytical Platform (Costech Instrument) at Jackson State University.
as reagent grade and used without further purification. Nuclear magnetic
resonance (NMR) spectra were recorded on a Varian Unity INOVA 500
FT-NMR. Chemical shifts for the samples were measured in DMSO-d6 and calibrated against the sodium salt of
3-(trimethylsilyl)propionic-2,2,3,3-d4 acid (TSP) as an external reference in a sealed capillary tube.
NMR data were processed and analyzed with MestReNova version 6.1.1-6384.
The IR spectra were recorded on a Perkin Elmer-Spectrum One Fourier
transform infrared spectrometer with KBr disks in the range of 4000–400
cm–1. The melting point was determined on a Mel-Temp
(Electrothermal 120 VAC 50/60 Hz) melting point apparatus and was
uncorrected. Mass spectral data were obtained at electrospray ionization
mass spectrometry (ESI-MS) positive mode on a TSQ Quantum GC (Thermo
Scientific). Elemental analysis was carried out using an ECS 4010
Analytical Platform (Costech Instrument) at Jackson State University.
6
Spectroscopic Characterization of Compounds
7
Nuclear Magnetic Resonance Characterization Protocol
8
Metabolite Profiling via GC-MS
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For metabolite profiling GC-MS is mostly the method of choice. Here we used GC hyphenated to triple quadrupole (Thermo Scientific TSQ Quantum GC™, Bremen, Germany), as previously described in detail (40 ).
9
Polar Fraction Analysis of Metabolites
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Polar fraction analysis were carried out following Valledor, et al. [84 (link)] protocol with some minor changes on a triple quad instrument (TSQ Quantum GC; Thermo). In brief, 1 µL of sample was injected, and GC separated into a HP-5MS capillary column (30 m 0.25 mm) (Agilent Technologies). Oven temperature was increased from 80 °C to 200 °C at a 3 °C per min rate and then reduced to 25 °C at 10 °C per min and maintained at 25 °C for 3 min, followed by 4 min of post-run conditions at 30 °C. Mass spectrometer was operated in electron impact (EI) mode at 70 eV in a scan range of m/z 40–600. The identification of metabolites was based on the spectral characteristics and GC retention times of each individual metabolite through its comparison with the retention times and spectral characteristics of standards available in our in-house library and in Golm Metabolome Database [86 (link)].
10
Gas Chromatography-Mass Spectrometry Protocol
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Gas chromatography coupled to mass spectrometer (TSQ QUANTUM GC, Thermofisher Scientific) was used for GC-MS analysis. Balance (0.01 g, sartorius company, Germany), solid-phase extraction device (Supelco company), and ultrasonic cleaner (Branson) were also used during the experiment.
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