Fecal <sup>1</sup>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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/12/2/148 |
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| author | Catherine L. J. Brown Hannah Scott Crystal Mulik Amy S. Freund Michael P. Opyr Gerlinde A. S. Metz G. Douglas Inglis Tony Montina |
| author_facet | Catherine L. J. Brown Hannah Scott Crystal Mulik Amy S. Freund Michael P. Opyr Gerlinde A. S. Metz G. Douglas Inglis Tony Montina |
| author_sort | Catherine L. J. Brown |
| collection | DOAJ |
| 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 (<i>n</i> = 10) and chicken (<i>n</i> = 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. |
| first_indexed | 2024-03-09T21:27:51Z |
| format | Article |
| id | doaj.art-b12d1dbc98d742acbd7bdf503295b91a |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-09T21:27:51Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-b12d1dbc98d742acbd7bdf503295b91a2023-11-23T21:05:09ZengMDPI AGMetabolites2218-19892022-02-0112214810.3390/metabo12020148Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline CorrectionCatherine L. J. Brown0Hannah Scott1Crystal Mulik2Amy S. Freund3Michael P. Opyr4Gerlinde A. S. Metz5G. Douglas Inglis6Tony Montina7Agriculture and Agri-Food Canada, Lethbridge Research and Development Centre, 5403-1st Avenue S, Lethbridge, AB T1J 4B1, CanadaSouthern Alberta Genome Sciences Centre, University of Lethbridge, Lethbridge, AB T1K 3M4, CanadaSouthern Alberta Genome Sciences Centre, University of Lethbridge, Lethbridge, AB T1K 3M4, CanadaBruker BioSpin Corporation, 19 Fortune Drive, Billerica, MA 01821, USADepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB T1K 3M4, CanadaSouthern Alberta Genome Sciences Centre, University of Lethbridge, Lethbridge, AB T1K 3M4, CanadaAgriculture and Agri-Food Canada, Lethbridge Research and Development Centre, 5403-1st Avenue S, Lethbridge, AB T1J 4B1, CanadaSouthern Alberta Genome Sciences Centre, University of Lethbridge, Lethbridge, AB T1K 3M4, CanadaAnalysis 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 (<i>n</i> = 10) and chicken (<i>n</i> = 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.https://www.mdpi.com/2218-1989/12/2/148<sup>1</sup>H-NMRmetabolomicsfecesintestineentericmicrobiome |
| spellingShingle | Catherine L. J. Brown Hannah Scott Crystal Mulik Amy S. Freund Michael P. Opyr Gerlinde A. S. Metz G. Douglas Inglis Tony Montina Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction Metabolites <sup>1</sup>H-NMR metabolomics feces intestine enteric microbiome |
| title | Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
| title_full | Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
| title_fullStr | Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
| title_full_unstemmed | Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
| title_short | Fecal <sup>1</sup>H-NMR Metabolomics: A Comparison of Sample Preparation Methods for NMR and Novel in Silico Baseline Correction |
| title_sort | fecal sup 1 sup h nmr metabolomics a comparison of sample preparation methods for nmr and novel in silico baseline correction |
| topic | <sup>1</sup>H-NMR metabolomics feces intestine enteric microbiome |
| url | https://www.mdpi.com/2218-1989/12/2/148 |
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