The procedure was performed according to Lehnhardt et al. [4 ], with minor changes. The cooled wort sample (5 mL) was transferred with 50 µL internal standard (2 mg/L p-fluorobenzaldehyde in ethanol) to a 20 mL headspace vial and stored in a cooled autosampler tray (17 °C). Extraction was performed using a CAR-PDMS-DVB fiber. First, the fiber was loaded with o-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) for 10 min at 40 °C. Afterward, the headspace of the sample was extracted for 30 min at 40 °C. Next, the fiber was injected with a 1/5-split at 270 °C into a GC (GC-Ultra 1300, Thermo Scientific Inc., Waltham, MA, USA) coupled to a single quad mass spectrometer (ISQ 7000, Thermo Scientific Inc., Waltham, MA, USA). The GC was equipped with a DB-5 column (length, 60 m; inner diameter, 0.25 mm; and film thickness, 0.25 µm; Thermo Scientific Inc., Waltham, MA, USA). The carrier gas used was helium (flow rate 1.85 mL/min). The starting temperature was held at 60 °C for 4 min, followed by heating at 5 K/min to a final temperature of 250 °C, which was maintained for 3 min. A full scan mode (m/z 35–350), with a dwell time of 0.02 s, was applied to the analysis. Each sample was analyzed in triplicate. Peak detection was performed in Xcalibur 3.1.66.10 (Thermo Scientific Inc., Waltham, MA, USA).
Isq 7000
Manufactured by Thermo Fisher Scientific
Sourced in United States, Italy
The ISQ 7000 is a single quadrupole mass spectrometer designed for routine quantitative and qualitative analysis. It features a robust vacuum system, high-performance ion optics, and a sensitive detector to provide reliable and reproducible results. The core function of the ISQ 7000 is to perform mass spectrometry analysis of various samples.
Automatically generated - may contain errors
Lab products found in correlation
Trace 1300, by Thermo Fisher Scientific (9 mentions)
Trace 1310, by Thermo Fisher Scientific (6 mentions)
Xcalibur 3, by Thermo Fisher Scientific (2 mentions)
Db 5 column, by Thermo Fisher Scientific (2 mentions)
Gc ultra 1300, by Thermo Fisher Scientific (2 mentions)
Hp innowax, by Agilent Technologies (2 mentions)
Butyric acid, by Merck Group (1 mentions)
Isovaleric acid, by Merck Group (1 mentions)
Valeric acid, by Merck Group (1 mentions)
Isobutyric acid, by Merck Group (1 mentions)
Propionic acid, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Trace 1300 gc, by Thermo Fisher Scientific (1 mentions)
Tracegold tg 5silms column, by Thermo Fisher Scientific (1 mentions)
Tg 5ms gc column, by Thermo Fisher Scientific (1 mentions)
Tg fame capillary column, by Thermo Fisher Scientific (1 mentions)
26 protocols using isq 7000
1
GC-MS Analysis of Volatile Compounds
2
GC-MS Analysis of Cecal SCFAs
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The concentrations of SFCAs in cecal content were detected using a gas chromatography-mass spectrometry (GC-MS) system (Trace 1300 and ISQ 7000, Thermo Scientific) (BioNovoGene, Suzhou, China). SCFA standards including acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid were purchased from Sigma-Aldrich (St. Louis, USA).
3
GC-MS Analysis of Rosemary Essential Oil
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The GC-MS analysis of the essential oil samples was carried out at the Department of Analytical Chemistry, Faculty of Science, Assiut University, to determine the volatile and semi-volatile chemicals. The extract of rosemary was made with isopropanol and ethyl acetate. The GC-MS analysis was performed with a Thermo Scientific TM TRACE 1300 coupled to an ISQ-7000 and a Thermo Scientific TM TG-6MB 5 ms (30 m×0.250 mm×1.00 m) column from Thermo Scientific. The temperature in the GC oven was kept at 100 °C for 15 min, then increased to 150 °C at a rate of 10 °C/min, and then to 200 °C at a rate of 5 °C/min. Furthermore, the temperature was raised to 250 °C at a rate of 5 °C per minute, and then to 280 °C at a rate of 5 °C per minute. Helium gas was used as the carrier, with a flow rate of 0.5 mL/min. The mass spectrometer was set to electron ionization mode, with a temperature of 320 °C for the ion source and 280 °C for the MS transfer line. The NIST 17 mass spectrum library (mainlib) was used to identify volatiles, and the results are represented as a percentage of the total GC area.
4
Gas Chromatography-Mass Spectrometry Analysis
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A Thermo Scientific Gas Chromatographer (Trace 1310) with electronic pressure control was used for the gas chromatography (GC) analysis, this apparatus was coupled to a simple quadrupole mass spectrometry detector ISQ 7000 (Thermo Fisher Scientific, Wal-than, MA, USA). Two columns were used: a DB-5ms column (5% phenylmethylpolysiloxane, 30 m × 0.25 mm i.d., 0.25 μm film thickness) and a HP-INNOWax column (polyethylene glycol, 30 m × 0.25 mm i.d., 0.25 μm film thickness). The column temperature was programmed from an initial temperature of 60 °C for 5 min, followed by a gradient of 2 °C/min to 100 °C, followed by a gradient of 3 °C/min to 150 °C, followed by a gradient of 5 °C/min to 200 °C. Finally, the temperature was increased at 15 °C/min to 250 °C and held for 5 minutes. Ultrapure helium at a flow rate of 1 mL/min was used as a carrier gas and 1 μL of the EO solution in CH2Cl2 (1% v/v) was injected (split 40:1) [20 (link),46 (link),49 (link)].
5
GC-MS Analysis of Derivatized Samples
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MOX + TMS derivatized samples were analyzed by GC-MS. 1 μL of derivatized sample was injected by autosampler into a Trace 1300 GC (ThermoFisher Scientific) fitted with a TraceGold TG-5SilMS column (ThermoFisher Scientific) operating under the following conditions: split ratio = 20:1, split flow = 24 μL/min, purge flow = 5 mL/min, carrier mode = Constant Flow, and carrier flow rate = 1.2 ml/min. The GC oven temperature gradient was as follows: 80 °C for 3 min, increasing at a rate of 20 °C/min to 280 °C, and holding at a temperature at 280 °C for 8 min. Between sample runs, the autosampler injection syringe was washed 3 times with methanol and 3 times with pyridine. Ion detection was performed by an ISQ 7000 mass spectrometer (ThermoFisher Scientific) operated from 3.90 to 21.00 min in EI mode (-70eV) using select ion monitoring (SIM). The mass spectrometer was tuned and calibrated daily. Samples were analyzed in a randomized order, and the pooled sample detailed above was analyzed at the beginning of, at a set interval (about every eight injections) during, and the end of the analytical run.
6
SCFA Quantification in Fecal Samples
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The concentrations of SCFAs were determined in the fecal supernatant using GC-MS (Thermo Trace 1300-Thermo ISQ 7000). Samples were homogenized for 1 min with 500 μL of water and 100 mg of glass beads, and then centrifuged at 4 °C for 10 min at 12,000 rpm. Then, 200 μL supernatant was extracted with 100 μL of 15% phosphoric acid and 20 μL of 375 μg/mL 4-methylvaleric acid solution as IS and 280 μL ether. Subsequently, the samples were centrifuged at 4 °C for 10 min at 12,000 rpm after vortexing for 1 min, and the supernatant was transferred into the vial prior to GC-MS analysis.
7
Determination of Fecal SCFA Concentrations
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The concentrations of fecal SCFAs were determined using a GC-MS system (TRACE 1300 Series GC System, Thermo Fisher Scientific Inc., Waltham, MA, USA) which was fitted with a capillary column Agilent HP-INNOWAX (30 m, 0.25 mm, 0.25 μm). Mass spectrometric detection of metabolites was performed on a Thermo Scientific™ ISQ™ 7000 GC-MS system. Single ion monitoring mode was used, with an electron energy of 70 eV. Fecal samples (0.1 g) were thawed, added in 2 ml of distilled water, and ultrasonically mixed for 20 min. Samples were extracted in 50 μL of 15% phosphoric acid with 10 μL of 75 μg/mL 4-methylvaleric acid solution and 140 μL ether, before centrifugation at 4 °C for 10 min at 12,000 × g. The supernatants were transferred into fresh sample vials before GC-MS analysis.
8
Enantiomeric Analysis by GC-MS
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For enantiomeric analysis, a gas chromatography (Trace 1310, Thermo Fisher Scientific, Waltham, MA, USA) coupled to a mass spectrometer (quadrupole) (ISQ 7000, Thermo Fisher Scientific, Waltham, MA, USA) was used. Analyses were performed on an enantioselective GC column (MEGA-DEX DMT-Beta, Mega, Legnano, MI, Italy) with 30 m of length, 0.25 m of internal diameter, and 0.25 μm of thick stationary phase (2.3 -diethyl-6-tert-butyldimethylsilyl-β-cyclodextrin). Sample preparation, amount injected, injection temperature, and partition radius were similar to those described for GC-MS. The carrier gas used was helium with a flow of 1.0 mL/min and a speed of 40 cm/s. The chromatographic run began maintaining the oven at 60 °C for 5 min, then the temperature was increased with a ramp of 2 °C/min up to 230 °C. finally, this temperature was maintained for 5 min. The calculation of the enantiomeric excess and elution order was carried out according to the procedures previously described by Morocho et al. [38 (link)].
9
Profiling Drought and Salt Stress Effects on Poplar Leaf Fatty Acids
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The leaves of poplar treated by two weeks of drought and salt stresses were freeze-dried, and gas chromatography-mass spectrometry (GC-MS) was used to analyze the LCFA and VLCFA contents. All above samples were immersed in 3 mL of reaction solvent containing 1 mL of n-hexane and 2 mL of 10% acetyl chloride-methanol for 2 h at 95 °C. Next, 6 mL of 6% potassium carbonate solution was added to the mixture. The n-hexane obtained from the above mixture was then removed by vacuum concentration. The isolated solvents were injected into a GC-flame ionization detector (Thermo Trace1300, Waltham, MA, USA) or GC-MS (Thermo ISQ7000, Waltham, MA, USA).
The column temperature was kept at 140 °C for 5min and then increased to 180 °C at 10 °C/min. Subsequently, it was raised to 210 °C at 4 °C/min and then increased to 310 °C for 30min at 10 °C/min. The flame ionization detector was applied to analyze the LCFA and VLCFA contents. Chromeleon7.0 software was applied to analyze individual LCFA or VLCFA, and the NIST 17 (https://www.sisweb.com/software/ms/nist.htm database (accessed on 10 May 2022)) was used to classify LCFA or VLCFA data.
The column temperature was kept at 140 °C for 5min and then increased to 180 °C at 10 °C/min. Subsequently, it was raised to 210 °C at 4 °C/min and then increased to 310 °C for 30min at 10 °C/min. The flame ionization detector was applied to analyze the LCFA and VLCFA contents. Chromeleon7.0 software was applied to analyze individual LCFA or VLCFA, and the NIST 17 (
10
Rumen Fluid Fatty Acid Analysis
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The extraction, methylation, and determination of medium and long-chain fatty acids in rumen fluid using the GC system (Trace1300, Thermo Fisher Scientific, Waltham, MA, USA) were determined using a GC hydrogen flame ionization detector (GC–MS), and automatic sampler (ISQ7000, Thermo Fisher Scientific, Waltham, MA, USA) as described by Guo et al. [21 (link)]. The fatty acid methyl ester was determined using a capillary column, and the concentration of long-chain fatty acids was detected using C19:0 as an internal standard. The chromatographic column was a DB-5ms capillary column, with specifications of 60 m × 0.25 mm × 0.25 μm. The conditions were as follows: injection volume: 1 μL; injection temperature: 260 °C; split ratio: 5:1; carrier gas: helium (99.999%); flow: 1.5 mL/min; column temperature: 140 °C for 5 min, to 180 °C at 10 °C/min, 3 min up to 210 °C at 2 °C/min and up to 280 °C at 5 °C/min, held for 15 min.
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