GC/MS data from the initial analyses[13] were used. Extracts were analyzed by GC/MS using an Agilent 5977 mass‐selective detector connected to an Agilent 7890B gas chromatograph and equipped with an Agilent ALS 7693 autosampler. HP‐5MS fused silica capillary columns (Agilent, 30 m×0.25 mm, 0.25 μm) were used. Injection was performed in splitless mode (250 °C injector temperature) with helium as the carrier gas (constant flow of 1.2 ml/min).[13] The additional samples were analyzed using an Agilent model 5975 mass‐selective detector connected to an Agilent 7890A gas chromatograph and equipped with an Agilent ALS 7683B autosampler equipped with an identical column. The temperature program started in all analyses at 50 °C, was held for 5 min, and then rose at a rate of 5 °C/min to 320 °C, before being held at 320 °C for 5 min.[13] GC/DD‐IR spectra were measured with a Dani Instruments DiscovIR IR‐detector coupled to an Agilent Technologies 7890B gas chromatograph equipped with an identical capillary column as used for GC/MS analyses. Components were identified by comparison of mass spectra and gas chromatographic retention index of commercial and in‐house databases, the latter obtained from authentic reference samples. Enantiomer determinations were performed using a column with a Hydrodex‐6‐TBDM (heptakis‐(2,3‐di‐O‐methyl‐6‐O‐t‐butyldimethylsilyl)‐β‐cyclodextrin) phase.
7890b gas chromatograph
Manufactured by Agilent Technologies
Sourced in United States
The 7890B gas chromatograph from Agilent Technologies is a laboratory instrument designed for the separation and analysis of complex chemical mixtures. It features a temperature-controlled oven, a sample injection system, and a detector to identify and quantify the components within a sample.
Automatically generated - may contain errors
Lab products found in correlation
5977a mass spectrometer, by Agilent Technologies (8 mentions)
5977b mass spectrometer, by Agilent Technologies (3 mentions)
Db 5ms column, by Agilent Technologies (3 mentions)
Hp 5ms capillary column, by Agilent Technologies (2 mentions)
7200 quadrupole time of flight mass spectrometer, by Gerstel (2 mentions)
Mps robotic autosampler, by Gerstel (2 mentions)
Db 5ms ui column, by Agilent Technologies (2 mentions)
Msd chemstation, by Agilent Technologies (2 mentions)
Db wax capillary column, by Agilent Technologies (2 mentions)
Hp 5ms, by Agilent Technologies (2 mentions)
Hp 5ms ultra inert column, by Agilent Technologies (2 mentions)
Hp 5ms column, by Agilent Technologies (2 mentions)
5977a mass selective detector, by Agilent Technologies (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Rt 2560 column, by Restek (1 mentions)
H2so4, by Honeywell (1 mentions)
Hydrochloric acid (hcl), by Honeywell (1 mentions)
Sodium hydroxide (naoh), by Honeywell (1 mentions)
Sodium chloride (nacl), by Honeywell (1 mentions)
R limonene, by Merck Group (1 mentions)
81 protocols using 7890b gas chromatograph
1
GC/MS and GC/DD-IR Analysis of Compounds
2
GC-MS Analysis of Organic Compounds
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An Agilent Technologies 7890B gas chromatograph coupled to a 7000C tandem mass selective detector operating in EI ionization mode was used for the analyses. The GC separation was achieved by a Phenomenex Zebron 30 m × 250 µm × 0.25-µm column with a (5%-phenyl)-methylpolysiloxane stationary phase. The injection temperature was set to 220°C and the injection volume was 2 µL. A pulsed-splitless injection was used at 20 psi for 0.75 min. The solvent delay was about 3 mins. The oven temperature was programmed as follows: isothermal at 100°C for 1 min, then ramped at 30°C/min up to 200°C, held for 0 min, ramped again at 15°C/min to 290°C, and final isothermal at 290°C for 3 min. The total chromatographic run adds up to 13.3 min. The transfer line was held at 280°C and the EI source at 230°C.
3
Fatty Acid Methyl Ester Derivatization and Analysis
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Extraction was carried out with chloroform, hexane (Honeywell, analysis grade, ≥99%), and methanol (Honeywell, HPLC grade). The solvents used for derivatization were H2SO4 (Honeywell, Puriss, 95–97%), BF3 (Sigma Aldrich, 14% in methanol), HCl (Honeywell, Puriss, ≥37%), NaCl (Carlo Erba, reagent grade), sodium hydroxide (NaOH, Honeywell, reagent grade, anhydrous pellets, ≥98%), and potassium hydroxide (KOH, Honeywell, reagent grade).
Qualification of the FAMEs methyl erucate, methyl cetalaicate and methyl brassidate were performed with the marine source analytical standard, polyunsaturated fatty acid mix no1 (Pufa n°1) and quantification standard for transfats (Restek-35629).
The derivatized fats were measured on an Agilent 7890B gas chromatograph (GC) coupled to an Agilent 5977B mass spectrometer (MS). The instrument was equipped with a Gerstel autosampler. A Restek Rt-2560 column (100 m × 0.25 mm ID with 0.20 µm film thickness) was used.
Qualification of the FAMEs methyl erucate, methyl cetalaicate and methyl brassidate were performed with the marine source analytical standard, polyunsaturated fatty acid mix no1 (Pufa n°1) and quantification standard for transfats (Restek-35629).
The derivatized fats were measured on an Agilent 7890B gas chromatograph (GC) coupled to an Agilent 5977B mass spectrometer (MS). The instrument was equipped with a Gerstel autosampler. A Restek Rt-2560 column (100 m × 0.25 mm ID with 0.20 µm film thickness) was used.
4
Terpenoid Profiling in Plant Tissues
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Approximately 0.3 g of fresh or dry materials was ground frozen in liquid nitrogen, vortexed and then extracted with 1.5 mL hexane using an ultrasonic cleaner for 0.5 h followed by incubation at 40°C for 1 h. Samples were then centrifuged at 12,000 rpm for 5 min, after which the resulting supernatants were pipetted into new 2 mL tubes. Next, 1 mL of hexane extract was pipetted into 1.5 mL vials for GC-MS analysis. The extracts were then analyzed using an Agilent 7890B Gas Chromatograph with a 5977B inert Mass Selective Detector (Agilent, United States). Helium was used as the carrier gas (1 mL/min), then separated on an HP-5MS column (30 m × 250 μm × 0.25 μm film thickness). For separation, the GC oven temperature was programmed for an initial temperature of 35°C for 5 min followed by an increase of 12°C/min to 300°C, which was held for 5 min. A NIST17/demo1 Mass Spectral Library was used for metabolite identification. The terpenoid compounds were identified by the mass spectral library, after which the main monoterpenoids investigated in this study, including α-pinene, camphene, myrcene, limonene, linalool, camphor, borneol and bornyl acetate, were further identified using their authentic standards. The contents were quantified based on bornyl acetate standard curves, and there were three biological replicates for each tissue.
5
Microwave-Assisted FAME Extraction and Quantification
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Fatty acid methyl esters (FAMEs) were prepared using the MARS 6 express 40 position microwave reaction system and quantified by gas chromatography (GC) as described previously.[27 ] Briefly, fatty acids were extracted from 50 mg flash frozen hepatic tissue using a microwave‐assisted preparation of FAMEs for GC analysis. Potassium hydroxide (10 mL, 2.5% w/v) in methanol and internal standard (ISTD) (tricasonaoic acid, 100 µL, 10 mg mL−1 in chloroform) were added for saponification, microwaved and heated to 130 °C, and held for 4 min. Methanolic acetyl chloride (15 mL, 5% v/v) was added for esterification, microwaved, heated to 120 °C in 4 min and held for 2 min. Pentane (10 mL) was added for fatty acid extraction and saturated sodium chloride (20 mL) was added to induce phase separation. FAMEs were measured using an Agilent 7890B gas chromatograph fitted with a flame ionization detector (FID). Analytes were separated using a CP‐Sil 88 capillary with a 100 m × 0.25 mm internal diameter × 0.2 µm film thickness column. Compounds were identified by comparing their retention times with FAME Supelco standards. Peak area analysis was conducted using Agilent OpenLAB CDS 2.1 Workstation. The content of each fatty acid (mg g−1 of tissue) was calculated according to the following equation:
6
Enzymatic Conversion of Terpenes and Aldehydes
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3-Carene, R-(+)-limonene, (+)-α-pinene and (Z)-dec-7-enal were obtained from Sigma-Aldrich. cis-3-Nonene was obtained from MP Biomedicals (Santa Ana, CA). Reaction mixtures consisted of 200 μL of the CYP6DE3 microsomal fraction, 40 μL of HF-CPR microsomal fraction, 100 mM sodium phosphate buffer pH 7.6, 1.5 mM NADPH (Sigma-Aldrich) and 21 mM of substrate in a total volume of 602 μL. Control reactions containing only HF-CPR were identical to the experimental reactions except that the reaction buffer was substituted for the CYP6DE3 microsomal fraction. Reactions were incubated in a capped 5 mL vial and rotated lengthwise at 30 °C in a FisherBiotech Hybridization Incubator (Thermo Fisher Scientific, Waltham, MA) for three hours. The reactions were terminated and extracted using pentane:ether (1:1). The extracts were analyzed by GC-MS on a HP-5 ms capillary column (Agilent) using an Agilent (Santa Clara, CA) 7890B gas chromatograph coupled to a 5977A mass spectrum detector. The instrument running parameters were: initial temperature of 40 °C with a one min hold, 5 °C/min to 240 °C and 15 °C/min to 300 °C with a 5 min hold. The MS detector was a single quadrapole with an electron ionization source and a molecular weight scanning range of 40 to 700 atomic mass units and an ionization potential of 70 eV.
7
Volatile Flavor Analysis of Highland Barley Flour
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Different samples of highland barley flour (300 mg) were placed in 20 mL headspace bottles. Ten microliters of 2-octanol was added as an internal standard. The volatile flavor compounds were analyzed using a 7890B gas chromatograph, 5977B mass spectrometer, and DB-Wax chromatographic column (30 m × 250 μm × 0.25 μm; Agilent Technologies Inc., Santa Clara, CA, USA). The GC conditions were: extraction temperature, 60 °C; preheating time, 15 min; extraction time, 30 min; resolution time, 4 min; splitless mode; and carrier gas, helium. The column flow rate was 1 mL/min, the sample inlet temperature was 250 °C, and the quadrupole temperature was 150 °C. The temperature rise program of the column box was 40 °C for 4 min, followed by 5 °C/min at 245 °C for 5 min. The mass spectrometry conditions included: ionization voltage, −70 eV; ion source temperature, 230 °C; transmission line temperature, 250 °C; mass scan range, m/z 20–400; and solvent delay, 0 min.
8
GC-MS Metabolomics Profiling Protocol
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The analysis was performed on an Agilent 7890B gas chromatograph connected to an Agilent 5977A MSD. Samples were run according to methods recommended for metabolome databases [5 (link)]. Briefly, 1 µL sample was injected in splitless mode with an Agilent 7693 autosampler injector into deactivated splitless liner (Agilent 5190-3167, splitless, single taper, glass wool). Separation was done on an Agilent 30 m DB5-MS column with a 10 m DuraGuard column. Oven temperatures were set to 60 °C for 2 min, and then headed with 10 °C min−1 up to 325 °C and set to hold for 5 min at 325 °C. Metabolites were assigned using the NIST 2.0 Library and verified by injection of pure compounds under the same chromatographic conditions.
9
GC-MS Derivatization and Analysis of Polar Metabolites
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Derivatization was performed using automated sample prep WorkBench instrument (Agilent Technologies, Santa Clara, CA, USA). Dried polar metabolites were dissolved in 60 μL of 2% methoxyamine hydrochloride in pyridine (ThermoFisher) and held at 40 °C for 6 h. After the reaction, 90 μL of MSTFA (N-Methyl-N-(trimethylsilyl) trifluoroacetamid) was added, and samples were incubated at 60 °C for 1 h. Derivatized samples were analyzed by GC-MS using a DB-35MS column (30 m × 0.25 mm i.d. × 0.25 µm) installed in an Agilent 7890B gas chromatograph (GC) interfaced with an Agilent 7200 Accurate-Mass Quadrupole Time-of-Flight (QTOF) mass spectrometer (MS), operating under electron impact (EI) ionization at 70 eV. Samples (1 μL) were injected in a splitless mode at 250 °C, using helium as the carrier gas at a flow rate of 1 mL/min. The GC oven temperature was held at 100 °C for 2 min and increased to 325 °C at 10 °C/min. GC/MS data processing was performed using Agilent Muss Hunter software. Relative metabolites abundance was carried out after normalization to internal standard d27 Myristic acid and cell number and statistical analyses were performed using MetaboAnalyst 4.0 [32 (link)].
10
GC/QTOF-MS Analysis of Metabolites
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GC/QTOF-MS data was collected using an Agilent 7890B gas chromatograph (GC) and an Agilent 7200 quadrupole time-of-flight (QTOF) mass spectrometer (MS), fitted with a Gerstel MPS Robotic Autosampler. 1 μL of the sample volume was injected in splitless mode onto a DB5ms column (30 m × 250 μm, 0.25 μm film thickness; Agilent Technologies, Catalog #:19091S-433). GC conditions were as follows: initial oven temperature 60°C (hold 1 minute), ramp at 25°C/minute to 300°C (hold 2.5 minutes), ramp at 120°C/minute to 60°C (hold 1 minute); the total run time was 16.1 minutes. The inlet, transfer line, chemical ionization (CI) source, and quadrupole temperatures were set to 280°C, 300°C, 150°C, and 150°C respectively. The emission current was set to 4.2 μA and data collection was in 2 GHz mode, mass range m/z 50–650. For all GC/QTOF-MS experiments, the sample injection order was randomized using the “RAND” function in Microsoft Excel. The same GC column was used for data acquisition shown in Figure 5 and Figure 6 , column trimming as part of routine maintenance was responsible for slightly shorter retention times shown in Figure 6 . The QTOF mass spectrometer was operated in negative chemical ionization (nCI) mode with methane as the reagent gas (1 mL/minute); in nCI mode, derivatized molecules undergo electron capture dissociation (ECD) and are detected as deprotonated (M-H) ions.
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