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Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction
Analysis of enteric microbiota function indirectly through the fecal metabolome has the potential to be an informative diagnostic tool. However, metabolomic analysis of feces is hampered by high concentrations of macromolecules such as proteins, fats, and fiber in samples. Three methods—ultrafiltrat...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875708/ https://www.ncbi.nlm.nih.gov/pubmed/35208222 http://dx.doi.org/10.3390/metabo12020148 |
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author | Brown, Catherine L. J. Scott, Hannah Mulik, Crystal Freund, Amy S. Opyr, Michael P. Metz, Gerlinde A. S. Inglis, G. Douglas Montina, Tony |
author_facet | Brown, Catherine L. J. Scott, Hannah Mulik, Crystal Freund, Amy S. Opyr, Michael P. Metz, Gerlinde A. S. Inglis, G. Douglas Montina, Tony |
author_sort | Brown, Catherine L. J. |
collection | PubMed |
description | Analysis of enteric microbiota function indirectly through the fecal metabolome has the potential to be an informative diagnostic tool. However, metabolomic analysis of feces is hampered by high concentrations of macromolecules such as proteins, fats, and fiber in samples. Three methods—ultrafiltration (UF), Bligh–Dyer (BD), and no extraction (samples added directly to buffer, vortexed, and centrifuged)—were tested on multiple rat (n = 10) and chicken (n = 8) fecal samples to ascertain whether the methods worked equally well across species and individuals. An in silico baseline correction method was evaluated to determine if an algorithm could produce spectra similar to those obtained via UF. For both rat and chicken feces, UF removed all macromolecules and produced no baseline distortion among samples. By contrast, the BD and no extraction methods did not remove all the macromolecules and produced baseline distortions. The application of in silico baseline correction produced spectra comparable to UF spectra. In the case of no extraction, more intense peaks were produced. This suggests that baseline correction may be a cost-effective method for metabolomic analyses of fecal samples and an alternative to UF. UF was the most versatile and efficient extraction method; however, BD and no extraction followed by baseline correction can produce comparable results. |
format | Online Article Text |
id | pubmed-8875708 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-88757082022-02-26 Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction Brown, Catherine L. J. Scott, Hannah Mulik, Crystal Freund, Amy S. Opyr, Michael P. Metz, Gerlinde A. S. Inglis, G. Douglas Montina, Tony Metabolites Article Analysis of enteric microbiota function indirectly through the fecal metabolome has the potential to be an informative diagnostic tool. However, metabolomic analysis of feces is hampered by high concentrations of macromolecules such as proteins, fats, and fiber in samples. Three methods—ultrafiltration (UF), Bligh–Dyer (BD), and no extraction (samples added directly to buffer, vortexed, and centrifuged)—were tested on multiple rat (n = 10) and chicken (n = 8) fecal samples to ascertain whether the methods worked equally well across species and individuals. An in silico baseline correction method was evaluated to determine if an algorithm could produce spectra similar to those obtained via UF. For both rat and chicken feces, UF removed all macromolecules and produced no baseline distortion among samples. By contrast, the BD and no extraction methods did not remove all the macromolecules and produced baseline distortions. The application of in silico baseline correction produced spectra comparable to UF spectra. In the case of no extraction, more intense peaks were produced. This suggests that baseline correction may be a cost-effective method for metabolomic analyses of fecal samples and an alternative to UF. UF was the most versatile and efficient extraction method; however, BD and no extraction followed by baseline correction can produce comparable results. MDPI 2022-02-05 /pmc/articles/PMC8875708/ /pubmed/35208222 http://dx.doi.org/10.3390/metabo12020148 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Brown, Catherine L. J. Scott, Hannah Mulik, Crystal Freund, Amy S. Opyr, Michael P. Metz, Gerlinde A. S. Inglis, G. Douglas Montina, Tony Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
title | Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
title_full | Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
title_fullStr | Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
title_full_unstemmed | Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
title_short | Fecal (1)H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
title_sort | fecal (1)h-nmr metabolomics: a comparison of sample preparation methods for nmr and novel in silico baseline correction |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875708/ https://www.ncbi.nlm.nih.gov/pubmed/35208222 http://dx.doi.org/10.3390/metabo12020148 |
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