A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Strategy for Nuclear-Magnetic-Resonance-Based Metabolomics of Human Feces
Tekijät: Lamichhane S, Yde CC, Schmedes MS, Jensen HM, Meier S, Bertram HC
Kustantaja: AMER CHEMICAL SOC
Julkaisuvuosi: 2015
Journal: Analytical Chemistry
Tietokannassa oleva lehden nimi: ANALYTICAL CHEMISTRY
Lehden akronyymi: ANAL CHEM
Vuosikerta: 87
Numero: 12
Aloitussivu: 5930
Lopetussivu: 5937
Sivujen määrä: 8
ISSN: 0003-2700
DOI: https://doi.org/10.1021/acs.analchem.5b00977
Tiivistelmä
Metabolomic analyses of fecal material are gaining increasing attention because the gut microbial ecology and activity have an impact on the human phenotype and regulate host metabolism. :Sample preparation is a crucial step, and in this study, we recommend a methodology for extraction and analysis of fresh feces by NMR-based metabolomics. The evaluation of extraction solvents showed that buffet extraction is a suitable approach to extract metabolic information in feces. Therefore, the effects of weight-to-buffer (Wf:Vb) combinations and the effect of sonication and freeze thaw cycles on the reproducibility, chemical shift variability, and signal-to-noise ratio (SNR) of the H-1 NMR spectra were evaluated: On the basis of our results, we suggest that fresh fecal extraction with a Wf:Vb ratio of 1:2 may be the optimum choice to determine the overall metabolite composition of feces. In fact, more than 60 metabolites have been assigned in the NMR spectra obtained from the fresh fecal buffer extract, and assignments of the lipophilic signals are also presented. TO our knowledge, some of the metabolites are reported here for the very first time employing H-1 NMR spectroscopy on human fecal extracts.
Metabolomic analyses of fecal material are gaining increasing attention because the gut microbial ecology and activity have an impact on the human phenotype and regulate host metabolism. :Sample preparation is a crucial step, and in this study, we recommend a methodology for extraction and analysis of fresh feces by NMR-based metabolomics. The evaluation of extraction solvents showed that buffet extraction is a suitable approach to extract metabolic information in feces. Therefore, the effects of weight-to-buffer (Wf:Vb) combinations and the effect of sonication and freeze thaw cycles on the reproducibility, chemical shift variability, and signal-to-noise ratio (SNR) of the H-1 NMR spectra were evaluated: On the basis of our results, we suggest that fresh fecal extraction with a Wf:Vb ratio of 1:2 may be the optimum choice to determine the overall metabolite composition of feces. In fact, more than 60 metabolites have been assigned in the NMR spectra obtained from the fresh fecal buffer extract, and assignments of the lipophilic signals are also presented. TO our knowledge, some of the metabolites are reported here for the very first time employing H-1 NMR spectroscopy on human fecal extracts.
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