Colon contents were collected after 8 weeks of FMT. The SCFAs were extracted from the mouse feces using 1:1 acetonitrile/water, derivatized using 3-nitrophenylhdyrazones, and analyzed using a Jasper HPLC coupled to a Sciex 4500 MD system (LipidALL Technologies Co., Ltd, Changzhou, China). Briefly, a Phenomenex Kinetex C18 column (100×2.1 mm, 2.6 µm) was used to separate the individual SCFAs, with a mobile phase A consisting of 0.1% formic acid aqueous solution and a mobile phase B consisting 0.1% formic acid/acetonitrile. Octanoic acid-1-13C1 (Sigma-Aldrich) and butyric-2,2-d2 (CDN Isotopes) were used as internal standards for the quantification (Li et al., 2019a (link)).
Octanoic acid 1 13c1
Manufactured by Merck Group
Sourced in United States
Octanoic acid-1-13C1 is a chemical compound that contains a carboxyl group (-COOH) and a carbon-13 isotope at the first carbon position. It is a type of organic acid that can be used as a labeling agent in various research and analytical applications.
Automatically generated - may contain errors
4 protocols using octanoic acid 1 13c1
1
Quantification of Gut Short-Chain Fatty Acids
2
Quantitative SCFA Analysis in Human Milk
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SCFAs were extracted from human milk using acetonitrile and derivatized using 3-nitrophenylhdyrazones. SCFAs were analyzed on a Jasper HPLC coupled to a Sciex 4500 MD system. Individual SCFAs were separated on a Phenomenex Kinetex C18 column (100 × 2.1 mm, 2.6 μm) using 0.1% formic acid in water as mobile phase A and 0.1% formic acid in acetonitrile as mobile phase B. Octanoic acid-1-13C1 purchased from Sigma-Aldrich and Butyric-2,2-d2 from CDN Isotopes were used as internal standards for quantitation.
3
Quantitative Analysis of SCFAs and MCFAs
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SCFAs and medium-chain fatty acids (MCFAs) were extracted from serum using acetonitrile:water (1:1) and derivatized using 3-nitrophenylhdyrazones. SCFAs and MCFAs were analyzed using a Thermo Fisher U3000 DGLC (Waltham, MA, USA) coupled to a Sciex QTRAP 6500 Plus (Framingham, MA, USA). In brief, individual SCFAs and MCFAs were separated on a Phenomenex Kinetex C18 column (100 × 2.1 mm, 2.6 µm; Torrance, CA, USA) using 0.1% formic acid in water as mobile phase A and 0.1% formic acid in acetonitrile as mobile phase B. Octanoic acid-1–13C1 purchased from Sigma-Aldrich (St. Louis, MO, USA) was used as an internal standard for quantitation.
4
SCFA Extraction and Quantitation in Mouse Feces
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Short-chain fatty acids (SCFAs) were extracted from mouse feces using acetonitrile: water (1:1) and derivatized using 3-nitrophenylhdyrazones. SCFAs were analyzed on a Jasper HPLC coupled to Sciex 4500 MD system. In brief, individual SCFAs were separated on a Phenomenex Kinetex C18 column (100 × 2.1 mm, 2.6 μm) using 0.1% formic acid in water as mobile phase A and 0.1% formic acid in acetonitrile as mobile phase B. Octanoic acid-1-13C1 purchased from Sigma-Aldrich and Butyric-2, 2-d2 from CDN Isotopes were used as internal standards for quantitation (Li et al., 2019 (link)).
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