An Agilent Technologies 7890A gas chromatography system coupled to an Agilent Technologies 5975c inert MSD quadrupole mass spectrometer (Agilent Technologies, Germany) and equipped with an Agilent Technologies 7697A Headspace automatic injector (Agilent Technologies, Germany) and an DB-FFAP capillary column (30 m × 0.25 mm i.d., 0.25 μm film thickness, Agilent Technologies, Germany) was used to perform an analysis of SCFAs from fecal samples. The headspace was maintained at 80°C with an incubation time of 30 min. The samples (1 mL) were injected in a splitless mode into the column at a temperature of 80°C. Helium was used as the carrier gas at a constant flow rate of 1 mL/min through the column. The initial oven temperature was 50°C, which was maintained for 1 min and then raised to 200°C at a rate of 10°C/min. The temperatures of the ion source and injector were 250°C. The mass detector system was operated in an electron impact (EI) mode with an ionization energy of 70 eV. The data of ions monitored were collected from m/z 33 to 200. A qualitative analysis was performed by the National Institute of Standards and Technology (NIST 11) MS library.
7890a gas chromatography system
Manufactured by Agilent Technologies
Sourced in United States, Germany
The 7890A gas chromatography system is a laboratory instrument designed for the separation and analysis of complex chemical mixtures. It is capable of efficiently separating and detecting a wide range of volatile and semi-volatile organic compounds. The system features advanced technology and precision engineering to provide reliable and accurate results.
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Lab products found in correlation
5975c inert msd quadrupole mass spectrometer, by Agilent Technologies (2 mentions)
7697a headspace automatic injector, by Agilent Technologies (2 mentions)
Db ffap capillary column, by Agilent Technologies (2 mentions)
Hp 5ms capillary column, by Agilent Technologies (2 mentions)
K alpha, by Thermo Fisher Scientific (2 mentions)
Fiehn library, by Agilent Technologies (1 mentions)
5975c msd, by Agilent Technologies (1 mentions)
Microfuge, by Eppendorf (1 mentions)
5975c msd single quadrupole mass spectrometer, by Agilent Technologies (1 mentions)
Masshunter workstation qualitative analysis software, by Agilent Technologies (1 mentions)
Dsc 214 polyma, by Netzsch (1 mentions)
Avance 2 400 mhz spectrometer, by Bruker (1 mentions)
7683b automatic liquid sampler, by Agilent Technologies (1 mentions)
Sp 2380, by Merck Group (1 mentions)
5975c mass spectrometer, by Agilent Technologies (1 mentions)
Hp 5ms column, by Agilent Technologies (1 mentions)
Ph meter, by Thermo Fisher Scientific (1 mentions)
Agilent 5975c gc msd, by Agilent Technologies (1 mentions)
11 protocols using 7890a gas chromatography system
1
GC-MS Analysis of Short-Chain Fatty Acids
2
GC-MS Analysis of Metabolites
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GC-MS analysis was performed using an Agilent 7890A Gas Chromatography system coupled with a 5975C MSD (Agilent Technologies, Santa Clara, CA, USA), which was also equipped with a PTV injector and a CTC autosampler. Separation was performed with a 30 m HP-5 ms UI (Agilent J&W) capillary column, with a film thickness of 0.25 μm and an i.d. of 0.25 μm. Back-flush elution was carried out in a 1.5 m deactivated column with a film thickness of 0.18 mm. The total run time was 30 min, followed by a 12 min back-flush run.
The initial oven temperature was set at 60 °C for 1 min and then increased to 300 °C at a 10 °C/min rate. It was then maintained at 300 °C for 6 min. The injection volume was set at 1 μL. The solvent delay was 6 min, while helium (99.999%) was used as the carrier gas at a flow rate of 3 mL/min. The injection system was performed in splitless mode, and the PTV injector temperature was increased from 270 °C to 350 °C.
MS was operated at electron impact ionization mode (EI; 70 eV), where the ion source temperature and transfer line temperature were set at 230 °C and 250 °C, respectively. All mass spectra were acquired in full scan mode between 50 and 600 amu. GAVIN was used to perform peak integration complementary to AMDIS for peak deconvolution and identification. Metabolite identification was based on Agilent Fiehn Library.
The initial oven temperature was set at 60 °C for 1 min and then increased to 300 °C at a 10 °C/min rate. It was then maintained at 300 °C for 6 min. The injection volume was set at 1 μL. The solvent delay was 6 min, while helium (99.999%) was used as the carrier gas at a flow rate of 3 mL/min. The injection system was performed in splitless mode, and the PTV injector temperature was increased from 270 °C to 350 °C.
MS was operated at electron impact ionization mode (EI; 70 eV), where the ion source temperature and transfer line temperature were set at 230 °C and 250 °C, respectively. All mass spectra were acquired in full scan mode between 50 and 600 amu. GAVIN was used to perform peak integration complementary to AMDIS for peak deconvolution and identification. Metabolite identification was based on Agilent Fiehn Library.
3
Galacturonate Metabolism in L. suebicus
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Cell extract of galacturonate-grown L. suebicus LCV1 (5.1 ± 0.4 g L–1) was incubated for 30 or 180 min at 30°C in a reaction mixture containing triethanolamine (TEA) buffer (100 mM, pH 7.6), 2.5 mM MgCl2.6H2O, 5 mM ATP, and 5 mM mannonate or no ATP and with 5 mM 6-phosphogluconate, unless stated otherwise. Samples were centrifuged (13,000 × g, Microfuge, ThermoFisher Scientific, Waltham, NC, United States) after incubation and the supernatant was collected; 100 μL of the supernatant was frozen at −80°C (U101 Innova freezer, Eppendorf, Hamburg, Germany) and freeze-dried over-night (Mini Lyotrap freeze-dryer, LTE Scientific Ltd., Greenfield, Oldham, United Kingdom). The supernatant was derivatized according to Niedenführ et al. (2016) (link) without addition of AAL-mix and analyzed using a 7890A gas chromatography system (Agilent, Santa Clara, CA, United States) coupled to a 5975C MSD single quadrupole mass spectrometer (Agilent, Santa Clara, CA, United States) according to de Jonge et al. (2011) (link); split ratio of 1: 50 for standards, split ratio of 1: 10 for samples. Identification of the peaks was done via MassHunter Workstation Qualitative Analysis software (Agilent, version B06.00) and comparison to the NIST Standard Reference Database (version 2.0).
4
Quantifying Short-Chain Fatty Acids in Fecal Samples
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The short-chain fatty acid (SCFA) analysis was used by a 7890A gas chromatography system (Agilent Technologies, Germany) coupled to a 5975c inert MSD quadrupole mass spectrometer, a 7697A headspace automatic injector, and a DB-FFAP capillary column (30 mm × 0.25 mm i.d., 0.25 μm film thickness). A portion of the 0.1 g homogenized fecal sample was mixed in 1 mL of 6% H3PO4 solution by ultrasound for 3 min and was then placed in an automatic headspace sampler at 80°C for 30 min. Subsequently, the samples (1 ml) were injected into the column at 80°C. Helium gas was used as a carrier for samples to pass through the column at a constant flow rate of 1 mL/min. The oven was preheated at 50°C for 1 min; then heated up to 200°C at the rate of 10°C/min. The temperature of the ion source and injector was set to 250°C. The mass detector system was operated in an electron impact (EI) mode with ionization energy of 70 eV. The data of ions monitored were collected from m/z 33 to 200. The short-chain fatty acids were quantified by the MS library of the National Institute of Standards and Technology (NIST 11).
5
Thermal and Physicochemical Analysis of Deep Eutectic Solvents
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The thermal properties of the prepared DESs were analyzed by means of ultralow temperature differential scanning calorimetry (DSC 214 Polyma, NETZSCH) with scan rates of 10 °C min−1 in a mixed gas condition (air/N2 = 4). The viscosity of each DES was measured with a RM 100 CP2000 PLUS viscometer (Lamy Rheology). Every measurement was conducted with a shear rate of 5 s−1 at 25 °C. X‐ray photoelectron spectroscopy (XPS; K‐alpha, Thermo VG Scientific) was used to investigate the core‐level electron states of the samples. The UV–vis spectrum of the charged DES electrolyte was ascertained with GENESYS 10s. For the analysis, the electrolytes were diluted with pure H2O. The Raman spectra were collected using an ARAMIS dispersive Raman spectrometer (Horiba Jobin Yvon) with an excitation laser wavelength of 514 nm. Zn deposition morphologies were characterized by scanning electron microscopy (SEM; Sirion, FEI Company). Solid‐state 13C NMR analyses were conducted using a Bruker AVANCE II 400 MHz spectrometer with a 9.4 T magnetic field. Hydrogen evolution was analyzed using an Agilent Technologies 7890A gas chromatography system.
6
Plasma Phospholipid Fatty Acid Analysis
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An analysis of plasma phospholipid fatty acids was performed at a commercial laboratory in Oslo, Norway (AS Vitas). Plasma stored at −80°C were thawed overnight at 4°C and vortexed for 5 sec. Dichloromethane/methanol was added to 100-µL plasma and 100-µL internal standard (1,2 diheptadecaonyl-sn-glycero-3-phosphatidylcholin). After shaking and centrifugation, the supernatants were transferred to new glasses and washed in a 0.9% NaCl solution. The lower phase was transferred to solid-phase extraction columns. Neutral lipids were washed out with dichloromethane/isopropanol and methyl tertiary butyl ether/formic acid. Phospholipids were eluted with methanol. After evaporation to dryness in a vacuum centrifuge, phospholipids were transmethylated with sodium methoxide and fatty acid methyl esters were extracted to hexane before gas chromatography analysis. Analysis was performed on a 7890A Gas Chromatography system with a split/split less injector, a 7683B automatic liquid sampler, and flame ionization detection (Agilent Technologies, Palo Alto, CA). Separation was performed on an SP 2380 (30 m×0.22 mm i.d.×0.25-µm film thickness) column (Supelco, Inc., Bellefonte, PA). The coefficient of variation was 4% for the main fatty acids and 6% for EPA and DHA (AS Vitas, Oslo). Plasma phospholipid fatty acids were expressed as µg/mL and in weight percentage.
7
Lipid Extraction and Fatty Acid Profiling
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Total lipids were extracted with methanol:chloroform:88% formic acid (2:1:0.1, v/v/v); the organic phase was separated by the addition of 0.5 volume of extraction buffer (1 M [w/v] KCl and 0.2 M [w/v] H 3 PO 4 ) followed by centrifugation for 3 min at 3000g. The organic phase was dried to completion under N gas, resuspended in 125 mL of chloroform, and 25 mL of lipid extracts was separated by thin layer chromatography on Silica Gel 60 plates (EMD Chemicals) using the following solvent system for neutral lipids: petroleum ether:diethyl ether:acetic acid (80:20:1 [v/v/v]). The identity of TAG was confirmed based on the separation of known lipid standards. Following the visualization of lipids by brief iodine staining, fatty acid methyl esters of each lipid as well as total cellular lipids were prepared in 1 M (v/v) methanolic HCl by heating the samples at 80°C for 20 min. The internatal standard was 5 mg of pentadecanoic acid. Following the phase separation with hexane and 0.9% (w/v) NaCl, the organic phase containing fatty acid methyl esters was completely dried under N gas, resuspended in 60 mL of hexane, and quantified by gas chromatography with flame ionization detection (7890A gas chromatography system, Agilent Technologies) using a temperature and run profile previously described by Zäuner et al. (2012) , with minor modifications.
8
GC-MS Analysis of Metabolite Derivatives
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Gas chromatography–mass spectrometry analysis was performed using a 7890A Gas Chromatography System (Agilent) with an HP-5 ms capillary column (30 m × 0.25 mm × 0.25 μm; Agilent J&W Scientific) interfaced with a 5975C mass spectrometer (Agilent). One microliter of di-O-isopropylidene, aldonitrile, or methyloxime derivatives was injected into a 270°C injection port in splitless mode. Helium flow was maintained at 0.88 mL/min. The column temperature was held at 80°C for 1 min, ramped at 20°C/min to 280°C and held for 4 min, then ramped at 40°C/min to 325°C. After a 5-min solvent delay, the mass spectrometer collected data in scan mode from mass/charge ratio (m/z) values of 100 to 500. Each derivative peak was integrated using a custom MATLAB function (34 (link)) to obtain mass isotopomer distributions (MIDs) for specific ion ranges.
9
Carbazole Quantification in Fungal Cultures
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After incubation, the filamentous fungi cultures were disintegrated for 5 min using Mixer Mill MM400 (Retsch, Germany) and 1 M HCl was added to pH 3. Carbazole and its metabolites were extracted with ethyl acetate (1:1, v/v). Ethyl acetate extracts were dehydrated using anhydrous sodium sulfate and then evaporated to dryness at 43 °C. After evaporation, each sample was resuspended in 2 mL ethyl acetate and analyzed by GC-MS. Phenylcarbazole was used as an internal standard in the quantitative analysis. The Agilent Technologies 7890A Gas Chromatography system equipped with a methyl polysiloxane HP 5MS column (30 m × 0.25 mm) and a 5975C Triple-Axis Detector (Agilent Technologies, USA) was used. The analysis was carried out using helium as a gas carrier at 1.2 mL min−1. In quantitative and qualitative methods, the oven temperature was set in the ranges of 100–300 and 80–300 °C, respectively. For the quantitative analysis, the ion at 167 m/z was used. The qualitative analyses were carried out at the mass range 100-400 m/z.
10
Fermentation Process Composition Analysis
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The pH and alcohol, total acid, reducing sugar, organic acid, and free sugar contents of the samples were monitored by periodic sampling at 24-h intervals throughout the fermentation process. Alcohol concentration was measured using a 7890A Gas Chromatography System (Agilent Technologies, Santa Clara, CA, USA) with a headspace sampler, a flame ionization detector (FID), and an HP-FFAP gas chromatography column (30 m; i.d. 0.32 mm; film thickness 0.50 μm). Ultra-pure nitrogen was used as the carrier gas at a flow rate of 16.41 mL/min. The injector temperature was 230°C. The column conditions were 10 min at 60°C with the FID at 230°C. pH was measured using a pH meter (Fisher Scientific, Pittsburgh, PA, USA) and the total acid content was estimated (as titratable acidity) after titration to pH 7.0 with 0.1 N NaOH. Acetic acid content (%) was then calculated based on the total acid content. The reducing sugar content was determined by the 3,5-dinitrosalicylic acid (DNS) method (25 ) using glucose as the standard. Detection was performed at 575 nm using an ultraspectrophotometer (Ultrospec 3300 pro, Biochrom Ltd., Cambridge, England).
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