Following treatment, metabolites were extracted by scraping cells in 80% methanol and vortexing. Extracts were spin-clarified, and supernatants were collected and dried by vacuum centrifugation at 4°C (Labconco). Dried supernatant pellets were resuspended in 15 ul of 10 mg/ml methoxylamine in pyridine (Sigma), incubated for 30 min at 37°C, and derivatized by silylation with 35 μl of N-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide (MTBSTFA, Sigma) for 1h at 70°C. Derivatized samples were spin clarified and 1 μl per sample was injected in splitless mode into an Agilent 5977B/7890B GC-MS with a DB-5ms capillary column with DuraGuard (Agilent Technologies), using chromatographic methods as previously described61 (link). Succinate was confirmed using 13C4-succinate (m+ 4) standard, and had a retention time of 14.1 min, quantifying ion of 289 + n, and qualifying ion 331 + n. Data were analyzed in batch format using Mass Hunter Quantitative Analysis software (Agilent Technologies). Data are shown as total ion counts normalized relative to total ion abundance and corrected for natural abundance and normalized to cell number upon plating.
Agilent 7890b 5977b
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 7890B-5977B is a gas chromatography-mass spectrometry (GC-MS) system. It is designed for the separation, identification, and quantification of complex mixtures of chemicals. The 7890B is a high-performance gas chromatograph, and the 5977B is a single quadrupole mass spectrometer. Together, they provide a powerful analytical tool for a wide range of applications, including environmental analysis, food and beverage testing, and pharmaceutical research.
Automatically generated - may contain errors
Lab products found in correlation
Mtbstfa, by Merck Group (3 mentions)
Duraguard, by Agilent Technologies (3 mentions)
Pyridine, by Merck Group (2 mentions)
Urea standard, by Merck Group (2 mentions)
Cp sil 88 for fame, by Agilent Technologies (2 mentions)
Db fatwax ui, by Agilent Technologies (2 mentions)
Masshunter quantitative analysis software, by Agilent Technologies (1 mentions)
Ms5977b, by Agilent Technologies (1 mentions)
Themis g2 300, by JEOL (1 mentions)
Fatty acid methylation kit, by Nacalai Tesque (1 mentions)
Db wax capillary column, by Agilent Technologies (1 mentions)
Masshunter software, by Agilent Technologies (1 mentions)
Hp 5ms capillary column, by Agilent Technologies (1 mentions)
10 protocols using agilent 7890b 5977b
1
GC-MS Analysis of Metabolite Extraction
2
Polar Metabolites Analysis in Islets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polar metabolites were extracted in 100 µl of 80% methanol (GC-grade, Thermo Optima) added to 100 µl of spent media from human or mouse islets (60 islets per 2ml) and lysed by sonication. Extracts were spin-clarified, and supernatants were collected and dried by vacuum centrifugation at 4°C (Labconco). Dried supernatant pellets were resuspended in 10 mg per ml of methoxylamine in pyridine (Sigma), incubated for 30 min at 37°C, and derivatized by silylation with 70 µl of N-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide (MTBSTFA, Sigma) for 1h at 70°C. After spin clarifying, 1 µl of derivatized sample was injected in splitless mode into an Agilent 5977B/7890B GC-MS with a DB-5ms capillary column with DuraGuard (Agilent Technologies), using chromatographic methods as previously described40 (link). Data are shown either as concentrations (ng/ml) using a urea standard (Sigma) calibration curve or as values relative to PBS-treated controls. Between sample differences were corrected for by a factor of median chromatographic peak area.
3
Polar Metabolites Analysis in Islets
4
Metabolic Profiling of C. beijerinckii
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The C. beijerinckii NCIMB 8052 strain was grown anaerobically at 37 °C in the modified MP2 medium containing 5 g/L of glucose and 1 g/L of 13C-labeled sodium formate as the carbon sources. The 2 mL grown cells were harvested when they reached OD600 of 2.5 and then hydrolyzed with 6 M HCl for 12 h at 105 °C. The samples were dried in a vacuum centrifuge and then derivatized with 100 μL of pyridine and 50 μL of N-methyl-N-[tert-butyldimethylsilyl]trifluoroacetamide (MTBSTFA, M − 108) for 1 h at 85 °C. Following filtration (0.22-μm pore size; Millipore), 1 μL of samples were injected into the GC-MS system (Agilent 7890B–5977B, Agilent, USA) equipped with a HP-5MS column (19091S–433UI, 30 m × 0.25 mm × 0.25 μm), the carrier gas was Helium. The mass spectrometer (Agilent MS-5977B) was operated in the electron impact (EI) mode at 70 eV. GC-MS data were analyzed as described previously [33 (link)].
5
Comprehensive Catalyst Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phase analysis of catalysts was performed by D8 Advance X-ray diffractometer using Cu-Kα radiation (λ = 1.5418 Å). JEOL-7100F scanning electron microscope (SEM) and double spherical aberration corrected transmission electron microscope (Titan Cubed Themis G2 300/Titan Cubed Themis G2 30) were adapted to perform the morphology, elemental distribution and three-dimensional visualization of tomographic reconstruction of catalysts. XPS measurements were performed on AXIS SUPRA, Kratos. Mass spectra was collected by GC-MS (Agilent 7890b/5977b).
6
Murine Adipose Fatty Acid Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The composition of fatty acids in murine epididymal adipose tissue and serum was measured by gas chromatography–mass spectrometry (GC/MS), Agilent 7890B/5977B (Agilent Technologies, Santa Clara, CA, USA). A total of 15 µg of epididymal adipose tissue and 25 µl of serum were methylated with a fatty acid methylation kit (nacalai tesque, Kyoto, Japan). The final product was loaded onto a Varian capillary column (DB-FATWAX UI; Agilent Technologies). The capillary column used for fatty acid separation was CP-Sil 88 for FAME (100 m × an inner diameter of 0.25 mm × membrane thickness of 0.20 μm, Agilent Technologies). The column temperature was maintained at 100°C for 4 min and then increased gradually by 3°C/min to 240°C and held there for 7 min. The sample was injected in split mode with split ratio 5:1. Each fatty acid methyl ester was detected in selected ion monitoring mode. All results were normalized to the peak height of the C17:0 internal standard (24 (link)).
7
Volatile Profiling of Yogurt Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flavor is one of the most important properties of yogurt products, and volatile components leading to off-flavors can cause the product to be unsatisfactory for the tastes of consumers [29 (link)]. The yogurt sample was added into a glass vial (10 mL). Volatile compounds were extracted using Head Space Solid Phase Micro Extraction (HS-SPME) technique, and separated using a DB-Wax column (30 m × 250 μm × 0.25 μm; J and W Scientific, Folsom, CA, USA). Desorption of the extracted volatiles was carried out using a GC-MS system (Agilent 7890B-5977B, Agilent Technologies, Santa Clara, CA, USA) by the splitless mode. Helium was used as carrier gas at a flow rate of 1.0 mL/min, and the MS was operated in scan mode. The oven temperature was held at 40 ℃ for 4 min (desorption period), increased to 250 ℃ with a rate of 5 ℃·min−1, and then held at 250 ℃ for 5 min. The total run time was 50 min. The NIST 2011 mass spectral library (Gaithersburg, MD, USA) was used to identify the volatile compounds.
8
GC-MS Analysis of Fecal SCFAs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fecal samples were sent to Shanghai Lu-Ming Biotech Co., Ltd. (Shanghai, China) for GC-MS analysis of SCFAs. Briefly, 20 mg freeze-dried samples were diluted with 400 μL ice-cold ethanol, incubated at −20 °C for 2 min, and ground with a grinder (60 Hz, 2 min). The samples were subject to ultrasonic treatment for 20 min in an ice-water bath. The impurities in supernatants were removed by centrifuging at 13,523 × g at 4 °C for 10 min. The obtained supernatants were then filtered through a 0.22 μm organic phase pinhole filter and transferred into a new 2 ml glass vial for GC (GAS chromatography) analysis on an Agilent 7890B-5977B gas chromatograph system (Agilent Technologies, CA, USA), in which a DB-WAX capillary column (30 m × 0.25 mm × 0.25 μm) (Agilent, Folsom, CA, USA) was used. The SCFAs in each sample were identified and quantified by a flame ionization detector. The data were analyzed using the Masshunter software (Agilent, USA). The concentration of each SCFA was evaluated with the linear regression equations from the corresponding standard curves.
9
GC-MS Analysis of Organic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The GC-MS data were obtained using an Agilent 7890B-5977B (Agilent, Santa Clara, CA, USA) system equipped with an HP-5 MS capillary column (30 m × 0.25 mm, 0.25 μm film thickness, Agilent J & W Scientific, Folsom, CA, USA). Helium was used as a carrier gas at a constant flow rate of 1.0 mL/min. Samples of 1 μL were injected in split mode at a ratio of 10:1 and at a constant temperature of 280 °C. The ion source and interface temperatures were set to 230 and 250 °C, respectively. The initial temperature of the column was 40 °C for 5 min, after which it increased to 280 °C at a rate of 10 °C/min and remained constant for 5 min. Mass spectra were taken at 70 eV, and the mass range was from 35 to 780 m/z. The solvent delay was 8.5 min.
10
GC/MS Analysis of Serum Fatty Acids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The composition of FAs in frozen serum samples was measured by GC/MS, Agilent 7890B/5977B (Agilent Technologies, Santa Clara, CA, USA). We metylated 25 μl of serum using a FA methylation kit (Nacalai Tesque, Kyoto, Japan), and loaded the final product onto a Varian capillary column (DB-FATWAX UI; Agilent Technologies). The capillary column used for FA separation was CP-Sil 88 for FAME (100 m × an inner diameter of 0.25 mm × membrane thickness of 0.20 μm, Agilent Technologies). We set the temperature in column at 100 °C for 4 min and then increased gradually by 3 °C/min to 240 °C and held there for 7 min. We injected the samples in split mode at a split ratio of 5:1. Each FA methyl ester was detected in the selected ion monitoring mode. All results were normalized to the peak height of the C17:0 internal standard [26 ].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!