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Biomolecules. 2019 Mar 28;9(4). pii: E121. doi: 10.3390/biom9040121.

Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry-Investigation of Pre-Analytic Stability.

Author information

1
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. gerhard.liebisch@ukr.de.
2
Nutritional Physiology, Technical University of Munich, 85354 Freising, Germany. josef.ecker@tum.de.
3
ZIEL-Institute for Food & Health, Technical University of Munich, 85354 Freising, Germany. josef.ecker@tum.de.
4
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. sebastian.roth@ukr.de.
5
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. Schweizer.sabine@gmx.net.
6
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. Veronika.Oettl@stud.uni-regensburg.de.
7
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. hans-frieder.schoett@isas.de.
8
ZIEL-Institute for Food & Health, Technical University of Munich, 85354 Freising, Germany. Hongsup.Yoon@tum.de.
9
Chair of Nutrition and Immunology, Technical University of Munich, 85354 Freising, Germany. Hongsup.Yoon@tum.de.
10
ZIEL-Institute for Food & Health, Technical University of Munich, 85354 Freising, Germany. dirk.haller@tum.de.
11
Chair of Nutrition and Immunology, Technical University of Munich, 85354 Freising, Germany. dirk.haller@tum.de.
12
Department of Hematology and Oncology, Internal Medicine III, University Hospital Regensburg, 93053 Regensburg, Germany. Ernst.Holler@ukr.de.
13
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. Ralph.Burkhardt@ukr.de.
14
Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany. silke.matysik@ukr.de.

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.

KEYWORDS:

LC-MS/MS; SCFA; feces; mass spectrometry; method validation

PMID:
30925749
DOI:
10.3390/biom9040121
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