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

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...

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Autores principales: Liebisch, Gerhard, Ecker, Josef, Roth, Sebastian, Schweizer, Sabine, Öttl, Veronika, Schött, Hans-Frieder, Yoon, Hongsup, Haller, Dirk, Holler, Ernst, Burkhardt, Ralph, Matysik, Silke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6523859/
https://www.ncbi.nlm.nih.gov/pubmed/30925749
http://dx.doi.org/10.3390/biom9040121
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author Liebisch, Gerhard
Ecker, Josef
Roth, Sebastian
Schweizer, Sabine
Öttl, Veronika
Schött, Hans-Frieder
Yoon, Hongsup
Haller, Dirk
Holler, Ernst
Burkhardt, Ralph
Matysik, Silke
author_facet Liebisch, Gerhard
Ecker, Josef
Roth, Sebastian
Schweizer, Sabine
Öttl, Veronika
Schött, Hans-Frieder
Yoon, Hongsup
Haller, Dirk
Holler, Ernst
Burkhardt, Ralph
Matysik, Silke
author_sort Liebisch, Gerhard
collection PubMed
description 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.
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spelling pubmed-65238592019-06-03 Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability Liebisch, Gerhard Ecker, Josef Roth, Sebastian Schweizer, Sabine Öttl, Veronika Schött, Hans-Frieder Yoon, Hongsup Haller, Dirk Holler, Ernst Burkhardt, Ralph Matysik, Silke Biomolecules Article 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. MDPI 2019-03-28 /pmc/articles/PMC6523859/ /pubmed/30925749 http://dx.doi.org/10.3390/biom9040121 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Liebisch, Gerhard
Ecker, Josef
Roth, Sebastian
Schweizer, Sabine
Öttl, Veronika
Schött, Hans-Frieder
Yoon, Hongsup
Haller, Dirk
Holler, Ernst
Burkhardt, Ralph
Matysik, Silke
Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability
title Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability
title_full Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability
title_fullStr Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability
title_full_unstemmed Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability
title_short Quantification of Fecal Short Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry—Investigation of Pre-Analytic Stability
title_sort quantification of fecal short chain fatty acids by liquid chromatography tandem mass spectrometry—investigation of pre-analytic stability
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6523859/
https://www.ncbi.nlm.nih.gov/pubmed/30925749
http://dx.doi.org/10.3390/biom9040121
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