Liebisch, Gerhard and Ecker, Josef and Roth, Sebastian and Schweizer, Sabine and Oettl, Veronika and Schoett, Hans-Frieder and Yoon, Hongsup and Haller, Dirk and Holler, Ernst and Burkhardt, Ralph and Matysik, Silke (2019) Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass SpectrometryInvestigation of Pre-Analytic Stability. BIOMOLECULES, 9 (4): 121. ISSN 2218-273X,
Full text not available from this repository. (Request a copy)Abstract
Short chain fatty acids (SCFAs) are generated by the degradation and fermentation of complex carbohydrates, (i.e., dietary fiber) by the gut microbiota relevant for microbe-host communication. Here, we present a method for the quantification of SCFAs in fecal samples by liquid chromatography tandem mass spectrometry (LC-MS/MS) upon derivatization to 3-nitrophenylhydrazones (3NPH). The method includes acetate, propionate, butyrate, and isobutyrate with a run time of 4 min. The reproducible (coefficients of variation (CV) below 10%) quantification of SCFAs in human fecal samples was achieved by the application of stable isotope labelled internal standards. The specificity was demonstrated by the introduction of a quantifier and qualifier ions. The method was applied to investigate the pre-analytic stability of SCFAs in human feces. Concentrations of SCFA may change substantially within hours; the degree and kinetics of these changes revealed huge differences between the donors. The fecal SCFA level could be preserved by the addition of organic solvents like isopropanol. An analysis of the colon content of mice either treated with antibiotics or fed with a diet containing a non-degradable and -fermentable fiber source showed decreased SCFA concentrations. In summary, this fast and reproducible method for the quantification of SCFA in fecal samples provides a valuable tool for both basic research and large-scale studies.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HUMAN FECES; SPECTROMETRY; VALIDATION; SAMPLES; SCFA; mass spectrometry; feces; LC-MS; MS; method validation |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 15 Apr 2020 09:05 |
| Last Modified: | 15 Apr 2020 09:05 |
| URI: | https://pred.uni-regensburg.de/id/eprint/27209 |
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