Eighty μL of the supernatant were transferred to an auto-sampler vial for gas chromatography–mass spectrometry (GC–MS) analysis. GC separation was conducted by Agilent 7890B GC System (Agilent Technologies, CA, United States). Mass spectrometry was performed with an EI source with selected ion monitoring using an Agilent 5977A mass spectrometer (Agilent Technologies, CA, United States). Ion source capillary temperature was 230°C.
Agilent 5977a mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 5977A mass spectrometer is a high-performance analytical instrument used for the identification and quantification of chemical compounds. It utilizes electron ionization (EI) technology to generate and analyze ions from various sample types. The 5977A model provides accurate mass measurements and sensitive detection, making it a versatile tool for a wide range of applications.
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Lab products found in correlation
Agilent 7890b gc system, by Agilent Technologies (2 mentions)
Agilent 7890b, by Agilent Technologies (2 mentions)
Agilent 7890 gas chromatography system, by Agilent Technologies (1 mentions)
Agilent 7890b gas chromatograph, by Agilent Technologies (1 mentions)
Db waxms column, by Agilent Technologies (1 mentions)
Hp 5msi, by Agilent Technologies (1 mentions)
Db 5 column, by Agilent Technologies (1 mentions)
6 protocols using agilent 5977a mass spectrometer
1
GC-MS Analysis of Supernatant
2
Intracellular Metabolite Isotopic Labeling
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Isotopic labeling of extracted intracellular metabolites was determined by GC-MS. The dried intracellular metabolites were first dissolved in 50 μl of 2% methoxylamine hydrochloride in pyridine and incubated at 37°C for 90 min on a heating block. Next, 80 μl of N-(tert-Butyldimethylsilyl)-N-methyl-trifluoroacetamide with 1% tert-butyldimethylchlorosilane (Thermo Fisher Scientific) was added and the samples were incubated for 30 min at 60°C. After overnight incubation at room temperature, the derivatized samples were briefly centrifuged and the clear liquid was transferred into GC vials for GC-MS analysis. GC-MS analysis was performed on an Agilent 7890B GC system equipped with a DB-5MS capillary column (30 m, 0.25 mm i.d., 0.25-μm phase thickness; Agilent Scientific), connected to an Agilent 5977A mass spectrometer operating under ionization by electron impact at 70 eV.
3
Metabolic Analysis of Macaque Gut in SIV
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An untargeted primary metabolic analysis of macaque gut tissue was performed by gas chromatography-time of flight (GC-TOF) mass spectrometry (MS). Ten to 20 mg of cryopreserved gut tissue was obtained from SIV-infected animals during the early stage at 60 h postinfection (n = 3) or during chronic stage at 10 weeks postinfection (n = 4) and from SIV-negative controls (n = 4). Tissue samples were homogenized in an extraction mixture of acetonitrile, isopropanol, and water (3:3:2) and centrifuged, and supernatants were evaporated. Dried pellets were dissolved in 50% acetonitrile, evaporated, and subjected to a two-step derivatization using methoximation and trimethylsilylation. The GC-MS analysis was performed using an Agilent 7890 gas chromatography system coupled to an Agilent 5977A mass spectrometer by the West Coast Metabolomics Center at UC Davis, as previously described (45 (link)). Overrepresentation analysis (ORA) was performed in Metabolites Set Enrichment Analysis (MSEA) software (http://www.metaboanalyst.ca ) using the metabolic pathways library from MSEA for all metabolites showing a 1.3-fold change.
4
Cheese Compositional and Aroma Analysis
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NIR spectrometer, type MPA (Bruker Optik GmbH, USA) was applied for the determination of the basic chemical composition of cheeses. Selected chemical parameters (dry matter, fat, salt, pH, proteins) were measured at the AgroBioTech SPU research center. The cheese samples were homogenized into a fine powder, then transferred to a Petri´s dish (type Duraplan) and placed in the apparatus. The NIR instrument was calibrated by O.K. SERVIS BioPro, Ltd. for hard cheeses in the range as follows: fat (5 -43.6%), NaCl (0.04 -2.85%), proteins (13.6 -37.10%), dry matter (24.2 -72.6%) and pH (4. 93 -6.35) .
The aroma profile was analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Agilent 7890B gas chromatograph coupled with an Agilent 5977A mass spectrometer (Agilent Technologies Inc., Palo Alto, CA, USA) was used. Separation was performed by a DB-WAXms column (30 m × 0.32 mm × 0.25 μm; Agilent Technologies) working with the temperature
The aroma profile was analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Agilent 7890B gas chromatograph coupled with an Agilent 5977A mass spectrometer (Agilent Technologies Inc., Palo Alto, CA, USA) was used. Separation was performed by a DB-WAXms column (30 m × 0.32 mm × 0.25 μm; Agilent Technologies) working with the temperature
5
GC-MS Analysis of Chemical Compounds
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GC–MS analysis was carried out using a 5977A Agilent mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) and a gas chromatograph (Agilent 7890B, Agilent Technologies) apparatus equipped with an Agilent HP-5MSi (30 m long and 0.25 mm i.d. with 0.25 µm film thickness) capillary column (95% dimethylpolysiloxane/5% diphenyl). The column temperature program was 60 °C for 5 min, with 3 °C/min increases to 180 °C, then 20 °C/min increases to 280 °C, which was maintained for 10 min. The carrier gas was helium at a 1 mL/min flow rate. Split mode injection (ratio 1:30) was employed. Mass spectra were taken over the m/z range of 30–650 with an ionizing voltage of 70 eV. MS identified the resulting individual compounds, and their identity was confirmed by comparison of their mass spectra with data available in the NIST 11 mass spectral library and the literature.
6
GC-MS Analysis of Organic Compounds
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GC–MS analysis was performed on a 7890B Agilent gas chromatograph including a DB-5 column (60 cm, 0.25 µ) and a 5977A Agilent mass spectrometer. 1 μL of diluted samples (with ethyl acetate) was injected into the injection site. The temperature program was scheduled as follows: the initial temperature of the oven was 40˚C (held for 7 min) and programmed to reach 140˚C with a rate of 10 ˚C /min, eventually reached 250˚C with a rate of 3˚C /min and held for 7 min at this temperature. Helium with 99.99% purity was utilized as a carrier gas (flow rate: 1 mL/min). Also, the ionization voltage of the detector was set at 70 eV. To determine the components, normal alkanes (C7–C21) were injected in the same manner to compare calculated retention indices with those in authentic references. For more accurate identification, the mass spectra of each compound were reconciled with the NIST database [34 ] and Adams's book [35 ].
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