The SCFA-concentrations were determined by means of diethyl ether extraction and capillary gas chromatography coupled to a flame ionization detector as described by De Paepe et al. (2017) (link) and Anderson et al. (2017) (link). Briefly, 1 ml aliquots were diluted 2× with 1 ml milli-Q water, and SCFA were extracted by adding approximately 400 mg NaCl, 0.5 ml concentrated H2SO4, 400 μl of 2-methyl hexanoic acid internal standard and 2 ml of diethyl ether before mixing for 2 min in a rotator and centrifuging at 3,000 g for 3 min. Upper layers were collected and measured using a GC-2014 capillary gas chromatograph (Shimadzu, Hertogenbosch, Netherlands), equipped with a capillary fatty acid-free EC-1000 Econo-Cap column (Alltech, Lexington, KY, United States), 25 m × 0.53 mm; film thickness 1.2 μm, and coupled to a flame ionization detector and split injector. One sample [donor 9, timepoint 2–0.05% (m/v) CMC] returned only zero values, presumably due to a technical error. This sample was therefore omitted prior to computational analyses.
Gc 2014 capillary gas chromatograph
Manufactured by Shimadzu
Sourced in Netherlands
The GC-2014 is a capillary gas chromatograph manufactured by Shimadzu. It is designed for the separation, identification, and quantification of volatile and semi-volatile compounds in a variety of samples. The GC-2014 features a compact design and advanced technology to provide efficient and reliable performance.
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SCFA Quantification by GC-FID
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Accurate SCFA Quantification by GC-FID
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The SCFA-concentrations were determined by means of diethylether extraction and capillary gas chromatography coupled to a flame ionization detector [25, (link)26] (link). Briefly, 1 mL aliquots were were diluted 2x with 1 mL mili-Q water and SCFA were extracted by adding approximately 400 mg NaCl, 0,5 mL concentrated H2SO4, 400 µL of 2-methyl hexanoid acid internal standard and 2 mL of diethyl ether before mixing for 2 min in a rotator and centrifuging at 3000 g for 3 minutes. Upper layers were collected and analysed using a GC-2014 capillary gas chromatograph (Shimadzu), equipped with a capillary fatty acid-free EC-1000 Econo-Cap column (Alltech), 25 m × 0,53 mm; film thickness 1,2 μm, and coupled to a flame ionization detector and split injector. The resulting SCFA-levels were visualized in R (version 4.1.0) by use of lineplots created using ggplot2 (v3.3.3).
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