The fatty acid composition was determined by conversion of oil to fatty acid methyl esters, prepared following the standard IUPAC method 2.301. Fatty acid compositions of sesame seed oils were analyzed by an Agilent Technologies 6890 N gas chromatograph (GC) equipped with a 30.0 m × 250 µm × 0.25 µm BPX-70 capillary column and detected using a flame ionization detector (FID). Samples (1 µL) were injected under the following conditions: the nitrogen flow rate was 1.0 mL/min, the oven was programmed from the set temperature of 170 to 210 °C at 2 °C/min, the split ratio was 50:1, the GC injection temperature was 250 °C, and the detector temperature was 300 °C.
6890n gas chromatograph
Manufactured by Agilent Technologies
Sourced in United States, Germany, Japan
The Agilent 6890N Gas Chromatograph is an analytical instrument designed for separating and identifying the individual components of a complex chemical mixture. It operates by heating the sample and using an inert carrier gas to transport the vaporized components through a column, where they are separated based on their different boiling points and interactions with the column material. The separated components are then detected and identified using various detectors, such as flame ionization or mass spectrometry.
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Lab products found in correlation
5973n mass spectrometer, by Agilent Technologies (7 mentions)
5975 mass selective detector, by Agilent Technologies (6 mentions)
Hp 5ms, by Agilent Technologies (6 mentions)
Hp 5ms capillary column, by Agilent Technologies (4 mentions)
Dxr raman microscope, by Thermo Fisher Scientific (4 mentions)
Hp 5ms column, by Agilent Technologies (3 mentions)
5975 mass spectrometer, by Agilent Technologies (3 mentions)
Db 5 column, by Hewlett-Packard (2 mentions)
Chromatof software, by Leco (2 mentions)
Agilent masshunter work station software, by Agilent Technologies (2 mentions)
5973 mass selective detector, by Agilent Technologies (2 mentions)
Zb 5ms, by Phenomenex (2 mentions)
Topspin software, by Bruker (2 mentions)
7683 autosampler, by Agilent Technologies (2 mentions)
5973b mass spectrometer, by Hewlett-Packard (2 mentions)
Sp 2560 column, by Merck Group (2 mentions)
Db 5ms column, by Agilent Technologies (2 mentions)
5973 mass spectrometer, by Agilent Technologies (2 mentions)
5975b mass selective detector, by Agilent Technologies (2 mentions)
5975c mass spectrometer, by Agilent Technologies (2 mentions)
110 protocols using 6890n gas chromatograph
1
Sesame Seed Oil Fatty Acid Composition
2
Quantitative Analysis of Baijiu Flavors
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As described previously [19 (link)], an Agilent (Santa Clara, CA, USA) 6890N gas chromatograph, fitted with a flame ionization detector, was employed to identify and quantify the major sweet compounds isolated (Tables S4 and S5 ). Baijiu samples (1 μL) with internal standard (IS5: amyl acetate, 174.24 mg·L−1) added were directly injected into the gas chromatograph in split mode (split ratio = 10:1). Nitrogen was used as the carrier gas at a constant flow rate of 2 mL/min, and a DB-Wax column (30 m × 0.25 mm i.d., 0.25 μm film thickness, J&W Scientific, Folsom, CA, USA) was used for separation. The column temperature was programmed as follows: initially 35 °C for 2 min; increased to 70 °C at 3.5 °C/min; increased to 180 °C at 5 °C/min; increased to 10 °C/min to 200 °C and held for 5 min. The injector and detector temperatures were set at 250 °C. A calibration curve was established for each standard compound, by injecting a dilution series, prepared in 50% (v/v) aqueous ethanol and diluted stepwise in a 1:1 ratio. All samples were tested in triplicate.
3
Quantifying Short Chain Fatty Acids
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We examined short chain fatty acids between C2:0 and C7:0. We were especially interested in the differences and changes of butyrate (C4:0) levels of the samples.
Stool samples from patients and healthy individuals were stored in small containers in -80°C. The patient samples were collected before and after FMT. Furthermore, native donor samples were compared to lyophilised samples.
Due to major differences between the concentrations of the samples, a qualitative analysis of the SCFA levels was carried out. 100 mg of the sample was diluted in 1 mL distilled water, followed by 10 minutes of vortexing. The samples were then incubated in room temperature for 10 minutes, then centrifuged with 10000 RPM (15000 g) for 10 minutes. The supernatant was collected, and mixed with the internal standard solution. The samples were then filtered (Low Protein Binding Hydrophilic LCR (PTFE) Membrane 0.2µm, Millex®, Tullagreen, Ireland) into GC tubes for the gas chromatography.
An Agilent Technologies 6890N gas chromatograph with a 5975 mass selective detector (Agilent, Waldbronn, Germany) was used for the analysis. Data analysis was performed using the GC/MSD CHEMSTATION (Version D.03.01, Agilent) software. Components were identified with the help of the NIST MS Search 2.0 library ( , ) and by spiking the samples with standards. C7:0 in its methyl ester form was used as an internal standard.
Stool samples from patients and healthy individuals were stored in small containers in -80°C. The patient samples were collected before and after FMT. Furthermore, native donor samples were compared to lyophilised samples.
Due to major differences between the concentrations of the samples, a qualitative analysis of the SCFA levels was carried out. 100 mg of the sample was diluted in 1 mL distilled water, followed by 10 minutes of vortexing. The samples were then incubated in room temperature for 10 minutes, then centrifuged with 10000 RPM (15000 g) for 10 minutes. The supernatant was collected, and mixed with the internal standard solution. The samples were then filtered (Low Protein Binding Hydrophilic LCR (PTFE) Membrane 0.2µm, Millex®, Tullagreen, Ireland) into GC tubes for the gas chromatography.
An Agilent Technologies 6890N gas chromatograph with a 5975 mass selective detector (Agilent, Waldbronn, Germany) was used for the analysis. Data analysis was performed using the GC/MSD CHEMSTATION (Version D.03.01, Agilent) software. Components were identified with the help of the NIST MS Search 2.0 library ( , ) and by spiking the samples with standards. C7:0 in its methyl ester form was used as an internal standard.
4
Headspace GC-TOF-MS Analysis
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After sampling for 10 min the fiber was desorbed for 1 min in an Optic injector port (ATAS GL Int., Zoeterwoude, NL) which was constantly kept at 250°C. Compounds were separated on a DB-5 column (10 m × 180 μm, 0.18 μm film thickness; Hewlett Packard) in a 6890N gas chromatograph (Agilent, Amstelveen, NL) with a temperature program set to 40°C for 1.5 min, increasing to 250°C at 30°C per min and 250°C for an additional 2.5 min. Helium was used as carrier gas, with the transfer column flow set to 3 mL per minute for 2 min, and to 1.5 mL per minute thereafter. Mass spectra were generated by electron ionization with 70 eV electrons at 200°C and collected with a Time-of-Flight MS (Leco, Pegasus III, St. Joseph, MI, United States), with an acquisition rate of 20 scans per second.
5
Detailed Analytical Techniques for Chemical Characterization
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Gas Liquid Chromatography (GLC) analyses were performed using a Dani GC 1000 instrument (Dani Instruments S.p.A., Cologno Monzese, Italy) equipped with a programmed temperature vaporizer injector and recorded with a Dani DDS 1000 data station. The following capillary columns were used: (i) Agilent J&W HP-5ms column (30 m × 0.25 mm i.d. × 0.25 μm); (ii) Agilent J&W DB-5 column (30 m × 0.25 mm i.d. × 1 μm); (iii) Alltech AT-35 FSOT column (30 m × 0.25 mm i.d. × 0.25 μm). Chiral-GLC analyses were carried out through a CyclodexB (30 m × 0.25 mm × 0.25 μm) or a Chiraldex G-TA (20 m × 0.25 mm × 0.25 μm) column. EI-MS spectra were recorded at 70 eV by GLC-MS and performed on an Agilent 6890 N gas-chromatograph interfaced with an Agilent 5973 N mass detector. Elemental analyses were acquired with an Elementar Vario Micro Cube in CHNS mode.
1H NMR spectra were recorded on a Varian Gemini 200 or a Bruker 400 MHz spectrometer using tetramethylsilane as an internal standard. 1H NMR spectral data of the synthetized compounds are reported inSupplementary Materials section .
1H NMR spectra were recorded on a Varian Gemini 200 or a Bruker 400 MHz spectrometer using tetramethylsilane as an internal standard. 1H NMR spectral data of the synthetized compounds are reported in
6
Urine Metabolite Profiling by GC-MS
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Urine samples require no preparation prior to freezing. Neat urine samples were lyophilized without further pretreatment. To the dried samples, we added 20 μl of 40 mg/ml methoxylamine hydrochloride in pyridine, and samples were agitated at 30°C for 30 min. Subsequently, 180 μl of trimethylsilylating agent N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) was added, and samples were agitated at 37°C for 30 min. GC–MS analysis was performed using a Agilent 6890 N gas chromatograph (Atlanta, GA, USA) interfaced to a time-of-flight (TOF) Pegasus III mass spectrometer (Leco, St. Joseph, MI, USA).8 (link)–10 (link) Automated injections were performed with a programmable robotic Gerstel MPS2 multipurpose sampler (Mülheim an der Ruhr, Germany). We performed initial peak detection and mass spectrum deconvolution with ChromaTOF software (version 2.25, Leco), and later exported samples to the netCDF format for further data evaluation with MZmine and XCMS. We identified samples with >50% certainty. If this degree of certainty was not met they were given the name (unknown) followed by a numeric number.
7
Quantifying Metabolic Profiles in Ferments
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To evaluate metabolic output, the relative concentrations of two varietal thiols, fifteen esters, six higher alcohols, four C6 compounds, six terpenes, and five fatty acids of all ferments were quantified following the method described in Knight et al. (2015 (link)). Varietal thiols (3MH, 3MHA) were quantified using an ethyl propiolate derivatization and analyzed on an Agilent 6890N gas chromatograph (Santa Clara, CA, USA) equipped with a 7683B automatic liquid sampler, a G2614A autosampler and a 593 mass selective detector as outlined in Herbst-Johnstone et al. (2013b (link)). Esters, alcohols, C6 compounds, terpenes, and fatty acids were quantified simultaneously using a HS-SPME/GC-MS method outlined in Herbst-Johnstone et al. (2013a (link)). Raw data was transformed with GCMSD Translator and peak integration was performed using MS Quantitative Analysis, both part of the Agilent MassHunter Workstation Software (Version B.04.00, Agilent Technologies).
8
Chemical Composition Analysis of Biopesticides
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A sample of all biopesticides was used to analyze the chemical composition using gas chromatography coupled to spectrometry.
The chemical composition of the oil was determined using an Agilent Technologies 7890 Gas Chromatograph with an Agilent Technologies 5975C mass spectrometer detector with a J&W 122-1545.67659 DB-5ms Column, at 325 °C and 60 m × 250 µm × 0.25 µm. Temperature ramp with start cycle at 40 °C and ramp up at 5 °C per min to 180 °C, then ramp up at 1 °C per min to 200 °C for 2 min and finally 25 °C per min to 300 °C held for 3 min, for a run time of 45 min. The injection volume was 1 µL and a split of 30:1, with He carrier gas 1 mL min−1, for which 20 µL of the sample was taken and diluted in 1 mL of dichloromethane, to inject 1 µL of the solution to the equipment.
To determine the chemical composition of the Hydrolate, Macerated, Infusion and Biol this was performed using the Agilent 6890 N Gas Chromatograph, with Agilent 5975B mass detector, with Agilent HP-5MS 5% Phenyl Methyl Siloxane Column 30 m × 0.25id × 0.5 µm film, initial oven temperature 40 °C for 5 min, 1.5 °C increment per minute up to 80 °C, 5 °C per minute up to 200 °C, 1 min at 200 °C. The analysis time was 59.67 min, a splitless mode injection port was used, and the initial temperature was 250 °C. The type of gas used was helium, and its flow rate was 1.0 mL min−1 with SPME injection.
The chemical composition of the oil was determined using an Agilent Technologies 7890 Gas Chromatograph with an Agilent Technologies 5975C mass spectrometer detector with a J&W 122-1545.67659 DB-5ms Column, at 325 °C and 60 m × 250 µm × 0.25 µm. Temperature ramp with start cycle at 40 °C and ramp up at 5 °C per min to 180 °C, then ramp up at 1 °C per min to 200 °C for 2 min and finally 25 °C per min to 300 °C held for 3 min, for a run time of 45 min. The injection volume was 1 µL and a split of 30:1, with He carrier gas 1 mL min−1, for which 20 µL of the sample was taken and diluted in 1 mL of dichloromethane, to inject 1 µL of the solution to the equipment.
To determine the chemical composition of the Hydrolate, Macerated, Infusion and Biol this was performed using the Agilent 6890 N Gas Chromatograph, with Agilent 5975B mass detector, with Agilent HP-5MS 5% Phenyl Methyl Siloxane Column 30 m × 0.25id × 0.5 µm film, initial oven temperature 40 °C for 5 min, 1.5 °C increment per minute up to 80 °C, 5 °C per minute up to 200 °C, 1 min at 200 °C. The analysis time was 59.67 min, a splitless mode injection port was used, and the initial temperature was 250 °C. The type of gas used was helium, and its flow rate was 1.0 mL min−1 with SPME injection.
9
Lipid Extraction and Fatty Acid Analysis
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Total lipids from synaptic membrane fractions were extracted following the method of Bligh and Dyer [40 (link)] with modifications [41 (link)]. Fatty acid methyl esters (FAMES) were prepared, identified, and quantified as previously described [4 (link)]. Briefly, FAMES were prepared from total lipid extracts by subjecting them to strong acid hydrolysis (16.6% HCl in methanol at 85 °C overnight). Total FAMES were quantified using an Agilent Technologies 6890N gas chromatograph (GC) with a flame ionization detector (FID), using 15:0 and 17:0 as internal standards. VLC-SFA were quantified using an Agilent Technologies 7890 GC [41 (link)] with a 5975C inert XL mass spectrometer (MS) detector (Agilent Technologies). The GC-MS was operated in the electron impact (EI) single ion monitoring (SIM) mode. The 28:0 and 30:0 response values were obtained by using the m/z ratios 438.4 and 466.5, respectively, along with m/z 74.1 and 87.1. Sample concentrations were determined by comparison to external standards, using 25:0 and 27:0 as internal standards. Multivariate ANOVA with Tukey’s post hoc was used to determine statistical significance.
10
Carbohydrate Derivatization and GC-MS Analysis
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The eluent for carbohydrate analysis needs a simple derivatization prior to GC-MS analysis. Briefly, 2% 0.2 mL sodium borohydride solution (dissolved in aqueous ammonia) was added into the eluent. After reaction for 20 min at 40 °C, 0.4 mL acetic acid, 0.3 mL methylimidazole and 1 mL acetic anhydride was added to the solution followed by another 10 min reaction. Finally, the carbohydrate derivatives were dissolved in 500 μL dichloromethane. The carbohydrate detection was performed on an Agilent 6890N gas chromatograph equipped with a MSD 5975 mass spectrometer (GC-MS) and electron-impact ionization (EI). A split/splitless-type injector was used for sample introduction. Chromatographic separation was carried out with a HP-5MS capillary column (30 m × 250 μm × 0.25 μm, Agilent Technology, CA, USA). The inlet temperature was 240 °C, and the oven temperature programs were as follows: the initial oven temperature was 140 °C (held for 0.5 min), ramped at 30 °C min–1 up to 190 °C (held for 5 min), and ramped at 2 °C min–1 up to 210 °C (held for 2 min). Helium was used as the carrier gas at a constant flow rate of 1.2 mL min–1. The MSD was operated in the electron impact ion (EI) mode with a source temperature of 230 °C. The electron energy was 70 eV and the filament current was 200 A.
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