Fitness for purpose of stabilized stool samples for bile acid metabolite analyses

Abstract Biobanks and cohort studies are increasingly utilizing chemical stabilizers to collect and store stool samples for downstream DNA-based microbiome analyses. While stabilizers permit ambient-temperature collection and storage of samples for gut microbiome studies, the use of the same sample...

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Autores principales:	Lorie Neuberger-Castillo, Wim Ammerlaan, Fay Betsou
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3fff349302dd433181cf71b1d95e5511
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spelling	oai:doaj.org-article:3fff349302dd433181cf71b1d95e55112021-12-02T15:51:12ZFitness for purpose of stabilized stool samples for bile acid metabolite analyses10.1038/s41598-021-86784-02045-2322https://doaj.org/article/3fff349302dd433181cf71b1d95e55112021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86784-0https://doaj.org/toc/2045-2322Abstract Biobanks and cohort studies are increasingly utilizing chemical stabilizers to collect and store stool samples for downstream DNA-based microbiome analyses. While stabilizers permit ambient-temperature collection and storage of samples for gut microbiome studies, the use of the same sample type for downstream metabolomics assays has not been explored. Microbiome-metabolomics analysis of fecal samples is increasingly getting attention to further elucidate the mechanisms by which the gut microbiota influences the host. In this study, we evaluated fitness-for-purpose of OMNIgene-GUT-collected stool samples for downstream metabolomics assays in the scope of fecal bile acids (BA) quantification. Biocrates Bile Acids Kit was used for the quantification of BA from eight healthy donors’ samples collected in (1) OMNIgene-GUT kit and (2) snap frozen in −80 °C in duplicates. A highly selective reversed phase LC–MS/MS analysis method in negative ion multiple reaction monitoring (MRM) detection mode was applied to determine the BA concentrations in each sample.Total fecal BA levels were detectable in OMNIgene-GUT-collected samples (range: 29.9–903.7 pmol/mg). Paired t-test confirmed that there was a significant difference in the total BAs between the OMNIgene-GUT and snap frozen samples (p < 0.05). Extractions from snap frozen samples resulted in higher concentrations of total BAs (range: 243.7–1136.2 pmol/mg). Qualitative differences between individual donors’ BA profiles were detectable using the two sample collection methods. No significant difference was found in the relative concentrations of primary (CA, CDCA) or secondary (DCA, LCA, UDCA) unconjugated BAs to the total BA concentrations in OMNIgene-GUT-collected samples as compared with the snap frozen samples (Wilcoxon-Mann–Whitney test, p > 0.05). Passing-Bablok method comparison and correlation analyis showed a high degree of correlation in the relative concentrations of CA, CDCA, DCA and LCA between OMNIgene-GUT and snap frozen samples. For these four bile acids, the two methods are comparable at an acceptability bias of 30%. We conclude that the OMNIgene-GUT-collected stool samples are fit-for-purpose for downstream fecal bile acids analysis.Lorie Neuberger-CastilloWim AmmerlaanFay BetsouNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lorie Neuberger-Castillo Wim Ammerlaan Fay Betsou Fitness for purpose of stabilized stool samples for bile acid metabolite analyses
description	Abstract Biobanks and cohort studies are increasingly utilizing chemical stabilizers to collect and store stool samples for downstream DNA-based microbiome analyses. While stabilizers permit ambient-temperature collection and storage of samples for gut microbiome studies, the use of the same sample type for downstream metabolomics assays has not been explored. Microbiome-metabolomics analysis of fecal samples is increasingly getting attention to further elucidate the mechanisms by which the gut microbiota influences the host. In this study, we evaluated fitness-for-purpose of OMNIgene-GUT-collected stool samples for downstream metabolomics assays in the scope of fecal bile acids (BA) quantification. Biocrates Bile Acids Kit was used for the quantification of BA from eight healthy donors’ samples collected in (1) OMNIgene-GUT kit and (2) snap frozen in −80 °C in duplicates. A highly selective reversed phase LC–MS/MS analysis method in negative ion multiple reaction monitoring (MRM) detection mode was applied to determine the BA concentrations in each sample.Total fecal BA levels were detectable in OMNIgene-GUT-collected samples (range: 29.9–903.7 pmol/mg). Paired t-test confirmed that there was a significant difference in the total BAs between the OMNIgene-GUT and snap frozen samples (p < 0.05). Extractions from snap frozen samples resulted in higher concentrations of total BAs (range: 243.7–1136.2 pmol/mg). Qualitative differences between individual donors’ BA profiles were detectable using the two sample collection methods. No significant difference was found in the relative concentrations of primary (CA, CDCA) or secondary (DCA, LCA, UDCA) unconjugated BAs to the total BA concentrations in OMNIgene-GUT-collected samples as compared with the snap frozen samples (Wilcoxon-Mann–Whitney test, p > 0.05). Passing-Bablok method comparison and correlation analyis showed a high degree of correlation in the relative concentrations of CA, CDCA, DCA and LCA between OMNIgene-GUT and snap frozen samples. For these four bile acids, the two methods are comparable at an acceptability bias of 30%. We conclude that the OMNIgene-GUT-collected stool samples are fit-for-purpose for downstream fecal bile acids analysis.
format	article
author	Lorie Neuberger-Castillo Wim Ammerlaan Fay Betsou
author_facet	Lorie Neuberger-Castillo Wim Ammerlaan Fay Betsou
author_sort	Lorie Neuberger-Castillo
title	Fitness for purpose of stabilized stool samples for bile acid metabolite analyses
title_short	Fitness for purpose of stabilized stool samples for bile acid metabolite analyses
title_full	Fitness for purpose of stabilized stool samples for bile acid metabolite analyses
title_fullStr	Fitness for purpose of stabilized stool samples for bile acid metabolite analyses
title_full_unstemmed	Fitness for purpose of stabilized stool samples for bile acid metabolite analyses
title_sort	fitness for purpose of stabilized stool samples for bile acid metabolite analyses
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3fff349302dd433181cf71b1d95e5511
work_keys_str_mv	AT lorieneubergercastillo fitnessforpurposeofstabilizedstoolsamplesforbileacidmetaboliteanalyses AT wimammerlaan fitnessforpurposeofstabilizedstoolsamplesforbileacidmetaboliteanalyses AT faybetsou fitnessforpurposeofstabilizedstoolsamplesforbileacidmetaboliteanalyses
_version_	1718385628710699008

Fitness for purpose of stabilized stool samples for bile acid metabolite analyses

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