SCFAs (acetate, propionate, and butyrate) in the colonic contents or fecal samples were quantified with gas chromatography. Briefly, 100 mg of samples were dissolved, homogenized, and centrifuged at 5400 rpm for 10 min. The supernatant was mixed with 0.2 mL deproteinized solution of 2-ethylbutyric acid in an ice-water bath for 30 min, then recentrifuged at 10,000 rpm for 10 min. The acquired supernatant was then kept in a 2-mL screw-cap vial and analyzed for the SCFA content using a gas chromatography system (Agilent Technologies, Wilmington, DE, USA).
Gas chromatography system
Manufactured by Agilent Technologies
Sourced in United States
The gas chromatography system from Agilent Technologies is an analytical instrument used for the separation, identification, and quantification of volatile and semi-volatile chemical compounds in a complex mixture. It functions by vaporizing the sample, separating the components using a specific column, and detecting the separated compounds through various detectors.
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Lab products found in correlation
5 protocols using gas chromatography system
1
Quantification of Gut Short-Chain Fatty Acids
2
Quantitative Analysis of Fecal SCFAs
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SCFAs were extracted and analyzed from each fecal samples, with some modifications as described previously (52 (link)). SCFAs, including acetate, propionate, and butyrate in fecal samples were quantified with gas chromatography. Briefly, 50 mg of fecal samples were weighed, dissolved, homogenized, and then centrifuged at 3, 000 × g for 5 min. The supernatant was adjusts pH to 2 to 3 with HCl, and then centrifuged at 12, 000 × g for 10 min, the last filtered through a 0.22 μm sterile membrane, kept in a 2 mL screw-cap vial, and then subjected for SCFAs analysis with an gas chromatography system (Agilent Technologies, USA).
3
Cecal VFA Analysis by GC
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Cecal VFAs were assayed by Gas Chromatography System (7890B, Agilent Technologies) according to previous studies (Yang et al., 2019 (link)). This analysis used a DB-FFAP, 30 m × 0.25 mm × 0.25 μm column (Agilent Technologies). Standards (acetic acid, propionic acid, butyrate, isobutyric acid, valerate, and isovalerate) and metaphosphoric acid were obtained from Aladdin Biochem Technology Co., Ltd. (Shanghai, China). The Gas Chromatography Analytical Conditions are listed as the follows: Carrier gas: N2. Sample injection volume:1 μL. Split ratio: 25:1. The initial column temperature was 90°C; this was held for 2 min then increased by 10°C min−1 to 200°C and was held for 5 min. In study, 2 g cecal contents were weighed into a 10 mL centrifuge tube and 2 mL of pre-cooled deionized water was added. The cecal contents were mixed, centrifuged at 4°C for 10 min (10,000 rpm) after stood at 4°C for 30 min. Took 1 mL of supernatant, added 0.2 mL 25% metaphosphoric acid (w/v), mixed well, centrifuged at 4°C for 10 min (15,000 rpm) after ice bath for 30 min. Centrifuge again at 4°C at 15,000 rpm for 10 min, then took 500 µL supernatant into a sample bottle for gas chromatography analysis.
4
Quantitative Analysis of Fecal SCFAs
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SCFAs were extracted and analyzed from each fecal samples, with some modifications as described previously (52 (link)). SCFAs, including acetate, propionate, and butyrate in fecal samples were quantified with gas chromatography. Briefly, 50 mg of fecal samples were weighed, dissolved, homogenized, and then centrifuged at 3, 000 × g for 5 min. The supernatant was adjusts pH to 2 to 3 with HCl, and then centrifuged at 12, 000 × g for 10 min, the last filtered through a 0.22 μm sterile membrane, kept in a 2 mL screw-cap vial, and then subjected for SCFAs analysis with an gas chromatography system (Agilent Technologies, USA).
5
Quantifying Short-Chain Fatty Acids in Fecal Samples
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Colonic contents or fecal samples were collected for SCFA (such as acetate, propionate, and butyrate) measurement via gas chromatography as previously described (26 (link)). In detail, 100 mg of samples from mice were dissolved, homogenized in ultrapure water, and suspension pH was adjusted to 2–3. The clear supernatant was collected post centrifugation at 5,400 rpm for 20 min. Zero-point-two milliliters deproteinized solution of 2-ethylbutyric acid was mixed with supernatant in an ice water bath for 30 min, followed by centrifugation at 10,000 rpm for 10 min. Acquired supernatant was transferred to a 2 ml screw-cap vial, and then subjected for SCFA analysis with Gas Chromatography System (Agilent Technologies, Wilmington, Delaware, USA).
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