Gc fid

Manufactured by Agilent Technologies

Sourced in United States

The GC-FID is a gas chromatography instrument equipped with a flame ionization detector (FID). It is designed to separate and detect a wide range of organic compounds in complex mixtures. The GC-FID provides reliable and sensitive analysis for various applications.

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44 protocols using gc fid

Fatty Acid Profiling of Yeast Cells

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Lipids were extracted from freeze-dried cells (around 20 mg) by transmethylation described by Browse et al. [18 (link)]. The methylated FA (FAMEs) were then analyzed using gas chromatography equipped with a flame ionization detector (GC-FID, Varian 3900, Walnut Creek, CA, USA) and a Varian FactorFour vf-23 ms column where the bleed specification at 260 °C is 3 pA (30 m, 0.25 mm, 0.25 μm).
The FAMEs were identified via comparisons with commercial standards (FAME32, Supelco, Bellefonte, PA, USA) and quantified using the internal standard, 100 μg of commercial dodecanoic acid (Sigma-Aldrich, St. Louis, MO, USA). Commercial standards of OCFAs (Odd Carbon Straight Chains Kit containing 9 FAs, OC9, Supelco, Bellefonte, PA, USA,) were converted into their FAMEs using the same method employed with the yeast samples. They were then analyzed by GC to identify the OCFAs from the yeast samples. For each data point, we used three biological replicates and calculated average and standard deviation values.

Abreu S., Park Y.K., Pires de Souza C., Vidal L., Chaminade P, & Nicaud J.M. (2022). Lipid Readjustment in Yarrowia lipolytica Odd-Chain Fatty Acids Producing Strains. Biomolecules, 12(8), 1026.

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Quantification of Higher Alcohols in Wines

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The fermentative component of wines was estimated by quantifying the major higher alcohols present in the finished wines (propan-1-ol, isopropanol, 2-methylpropanol, butan-1-ol, butan-2-ol, 2-methylbutanol, 3-methylbutanol). Individual alcohols were determined by GC-FID (Varian, Inc.), using a direct 1/1,000 split injection of 1 μL of the sample into a CP-WAX 50 × 32 capillary column in the presence of octan-2-ol as internal standard. A temperature programme from 25 to 240°C was used in the column oven and maintained at a constant temperature of 25°C for 5 min, followed by an increase of 5°C·min⁻¹ from 5 to 12 min and 15°C·min⁻¹ from 12 to 24 min. The injector temperature was fixed at 240°C and the temperature of the flame ionization detector was set at 250°C. The methodology had been described previously by the Office International de la Vigne et du Vin (32 ). All the higher alcohols were integrated based on their area in the chromatogram and transformed into concentrations expressed in mg·L⁻¹, before being summed to obtain the fermentative component.

Coelho C., Gougeon R.D., Perepelkine L., Alexandre H., Guzzo J, & Weidmann S. (2019). Chemical Transfers Occurring Through Oenococcus oeni Biofilm in Different Enological Conditions. Frontiers in Nutrition, 6, 95.

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Quantifying Carbon Tetrachloride by GC-FID

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CCl₄ was quantified using a CP 3800 gas chromatograph connected to a flame ionization detector (GC-FID; Varian, Palo Alto, CA, USA). Headspace aliquots (500 μL) were collected from sealed Hungate culture tubes with a gastight 1750 syringe (Hamilton, Franklin, MA, USA) and injected onto the GC column (CP-Sil 5 CB, length 15 m; Varian). Separation of volatile compounds was achieved by isothermal elution at 30 °C for 1 min, followed by a linear temperature gradient (20 °C min⁻¹) up to 220 °C. Injector and detector were maintained at 220 °C (splitless mode) and 300 °C, respectively, with nitrogen (N₂) as the make-up gas (Linde Gas). Peak areas were analyzed with Galaxie Workstation software (Varian) and expressed as percentage of CCl₄ compared to an abiotic reference tube prepared under the same conditions.

Penny C., Gruffaz C., Nadalig T., Cauchie H.M., Vuilleumier S, & Bringel F. (2015). Tetrachloromethane-Degrading Bacterial Enrichment Cultures and Isolates from a Contaminated Aquifer. Microorganisms, 3(3), 327-343.

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Fatty Acid Profiling of Infiltrated Leaves

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Total lipids extracted from the infiltrated leaves were esterified to determine fatty acid composition. To perform fatty acid methyl esterification (FAME), HCl methanolic acid (1 N) was added to the samples and the mixture was heated for 2 h at 85°C in a water bath. After cooling to room temperature, 1 mL KCl and 1 mL hexane were added for phase separation. The hexane organic phase containing FAMEs was collected and dried under nitrogen gas at 40°C. Each FAME sample was resuspended in 1 μL of hexane and analyzed by gas chromatography coupled with a flame ionization detector (GC-FID, Varian). A capillary column (DB-23; 30 m × 0.32 mm I.D., 0.25 μm) with helium as carrier gas (flow rate of 1.5 mL/min) was used. The injection temperature was set at 150°C, which was ramped after 3 min to 240°C at 6°C/min; the detection temperature was 300°C. The retention time for each fatty was determined based on comparison to the Supelco FAME mix standards, and quantified relative to the C17:0 internal standards. Each sample was analyzed in triplicate.

Behera J.R., Rahman M.M., Bhatia S., Shockey J, & Kilaru A. (2021). Functional and Predictive Structural Characterization of WRINKLED2, A Unique Oil Biosynthesis Regulator in Avocado. Frontiers in Plant Science, 12, 648494.

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Anaerobic Bacterial Cultivation and Diffusive Sampling

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All the tested LAB strains from A. mellifera were incubated anaerobically for 7 days in Pollen media (50 ml). Bacterial cultivations were performed separately in one anaerobic jar for each strain. The diffusive sampler was attached onto the inner side of the jar's lid. A diffusive sampler having uptake rates that fully agree with the theories behind diffusive sampling was used for sampling and analysis of formic and acetic acids (Ferm 2001, www.diffusivesampling.ivl.se). Analysis was made using ion chromatography with a gradient eluent generator (DIONEX ICS 2000). Diffusive sampling of other organic vapours was made with tube‐typed sorbent tubes (PerkinElmer, Waltham, MA). For sampling of benzene, toluene, n‐octane, ethylbenzene, m‐, p‐xylene, o‐xylene and n‐nonane, Carbopack B (Sigma‐Aldrich) was used as an adsorbent. The pollutants were analysed via thermal desorption (ATD‐400, PerkinElmer) and gas chromatography with a flame ionisation detector (GC‐FID, Varian3800). More samplers could be analysed when Tenax TA was used as sorbent, and analysis was made with thermal desorption (Markes, Frankfurt, Germany) and GC–MS (Agilent Technologies). Experimentally determined uptake rates were used for the thermally desorbed hydrocarbons.

Olofsson T.C., Butler È., Markowicz P., Lindholm C., Larsson L, & Vásquez A. (2014). Lactic acid bacterial symbionts in honeybees – an unknown key to honey's antimicrobial and therapeutic activities. International Wound Journal, 13(5), 668-679.

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Monosaccharide Composition Analysis of EPS

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Monosaccharide composition was determined according to the protocol described by Kamerling et al. [77 (link)] and modified by Montreuil et al. [78 ]. The EPS solution was firstly hydrolyzed for 4 h at 100 °C in methanol/HCl 3 N (Merck, Lyon, France). The obtained methyl glycosides were then converted to trimethylsilyl derivatives with the use of N,O-bis(trimethylsilyl)trifluoroacetamide and trimethylchlorosilane (BSTFA: TMCS) (Merck, Lyon, France) (99:1). Per-O-trimethylsilyl methyl glycosides formed were quantified using gas chromatography (GC-FID, Agilent Technologies, Les Ulis, France). The myo-inositol was used as the internal standard, as previously described [52 (link)].

Migaou M., Macé S., Maalej H., Marchand L., Bonnetot S., Noël C., Sinquin C., Jérôme M., Zykwinska A., Colliec-Jouault S., Maaroufi R.M, & Delbarre-Ladrat C. (2024). Exploring the Exopolysaccharide Production Potential of Bacterial Strains Isolated from Tunisian Blue Crab Portunus segnis Microbiota. Molecules, 29(4), 774.

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Fecal Biomarker Profiling Protocol

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Fecal pH was measured with an electrode-fitted pH meter LAQUAtwinB-712 (HORIBA, Kyoto, Japan) after suspending fecal samples in Milli-Q water and sterilizing at 85 °C for 15 min for sterilization of fecal samples.
Fecal SCFAs (acetate, propionate, n-butyrate, iso-butyrate, n-valerate, iso-valerate, n-caproic acid) in ethyl acetate extract were determined by gas chromatography system GC-FID (7890B, Agilent Technologies, Santa Clara, CA, USA) and a DB-WAXetr column (30 m, 0.25 mm id, 0.25 μm film thickness, 1.2 mL/min) as described previously [42 (link)].
Commercially available ELISA kits were used to determine fecal secretory immunoglobulin A (sIgA: Human IgA ELISA Core Kit, LABISKOMA, Seoul, Republic of Korea) and calprotectin (IDK^® Calprotectin MRP 8/14 ELISA kit, Immundiagnostik AG, Bensheim, Germany).

Hiraku A., Nakata S., Murata M., Xu C., Mutoh N., Arai S., Odamaki T., Iwabuchi N., Tanaka M., Tsuno T, & Nakamura M. (2023). Early Probiotic Supplementation of Healthy Term Infants with Bifidobacterium longum subsp. infantis M-63 Is Safe and Leads to the Development of Bifidobacterium-Predominant Gut Microbiota: A Double-Blind, Placebo-Controlled Trial. Nutrients, 15(6), 1402.

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Comprehensive Lipid Extraction and Analysis

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The total lipids from the whole body homogenates, liver samples, fillet samples, and diet samples were extracted according to Folch et al. [42 ] with modifications as in Torno et al. [22 (link)]. Samples were neutralized using potassium hydroxide (0.1 M) followed by the addition of the Folch reagent and subsequent centrifugation for 10 min at 2000× g to isolate the fatty acid methyl esters (FAMEs). The organic phase was collected to perform a second extraction with potassium hydroxide and the Folch reagent. After subsequent centrifugation (5 min at 2000× g) and drying of samples under a N₂ flux, the re-dissolved FAME samples were injected into a 7820A Agilent gas chromatograph with flame ionization detector (GC-FID; Agilent Technologies, Santa Clara, CA, USA). A FAME-standard was used for the identification of the retention times of the individual FAMEs. The fatty acid composition was calculated as percentage of single FAME relative to total FAMEs. The internal standard 13:0 methyl ester was used to calculate FAs as % DM of diet.

Torno C., Staats S., Fickler A., de Pascual-Teresa S., Soledad Izquierdo M., Rimbach G, & Schulz C. (2019). Combined effects of nutritional, biochemical and environmental stimuli on growth performance and fatty acid composition of gilthead sea bream (Sparus aurata). PLoS ONE, 14(5), e0216611.

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Quantification of Organic Acids and Gases

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To measure organic acids, samples were treated with BaCl₂ and OnGuard Ag cartridges (Dionex) to remove the high level of sulfate and chloride, and the pH raised prior to their measurement by ion chromatography.
S(-II) was determined with Cline method, by reaction with N,N-Dimethyl-p-phenylenediamine. Absorbance was measured at 664 nm using a spectrophotometer40 . Fe(II) were determined by reaction with ferrozine. Absorbance was measured at 562 nm using a spectrophotometer41 .
Dissolved gases (H₂ and CO₂) were measured with a GC-FID (Agilent Technologies), using a headspace equilibration method.

Bagnoud A., Chourey K., Hettich R.L., de Bruijn I., Andersson A.F., Leupin O.X., Schwyn B, & Bernier-Latmani R. (2016). Reconstructing a hydrogen-driven microbial metabolic network in Opalinus Clay rock. Nature Communications, 7, 12770.

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Quantitative Analysis of Leaf Wax Composition

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Forty-day-old plants of OE and WT grown in the same pot containing soil were treated with or without 200 mM NaCl for 30 days. Leaf discs with same area (6.872 cm²) were sampled from leaves with comparable age and position of OE and WT plants and waxes were extracted from leaf surface with chloroform for 30 s with two repeats. The resultant extracts were dried under a stream of nitrogen and were dissolved in 100 µL of chloroform. Wax constituents containing 10 mg tetracosane internal standard were converted to their trimethylsilyl ethers and esters by adding 20 µL of N, N-bis-trimethylsilyltrifluoroacetamide and 20 µL of pyridine to the extracts and incubating the mixture for 40 min at 70 °C. The fatty acid derivatives were quantitatively analyzed by GC-FID (Agilent Technologies, Santa Clara, CA, USA) and GC-MS (Agilent gas chromatograph coupled to an Agilent 5973 N quadruple mass selective detector) at the Instrumental Analysis Center of Shanghai Jiao Tong University.

Liu N., Chen J., Wang T., Li Q., Cui P., Jia C, & Hong Y. (2019). Overexpression of WAX INDUCER1/SHINE1 Gene Enhances Wax Accumulation under Osmotic Stress and Oil Synthesis in Brassica napus. International Journal of Molecular Sciences, 20(18), 4435.

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