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Metabolites 2018, 8(4), 75; https://doi.org/10.3390/metabo8040075

Gas Chromatography Mass Spectrometry (GC-MS) Quantification of Metabolites in Stool Using 13C Labelled Compounds

1
Institute of Biosensor Technology, University of the West of England, Bristol BS16 1QY, UK
2
Indigo Science Ltd., Bristol BS7 9JS, UK
3
Givaudan UK Ltd., Ashford TN24 0LT, UK
*
Authors to whom correspondence should be addressed.
Received: 21 September 2018 / Revised: 24 October 2018 / Accepted: 29 October 2018 / Published: 31 October 2018
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Abstract

It has become increasingly important to qualitatively and quantitatively assess the volatile metabolites in a range of bodily fluids for use in monitoring health. There has been relatively little work on the quantitative analysis of compounds, particularly with respect to the effects of ethnicity or geographic location. A novel method for the quantification of compounds in stool using 13C labelled compounds as internal standards is presented. Using thermal desorption gas chromatography mass spectrometry, stool samples from 38 healthy volunteers were analysed. The 13C labelled compounds, acetone, ethyl butanoate, ethanoic acid, butanoic acid, 3-methylbutanoic acid, and indole, were added as internal standards. This process mimics the solubility characteristics of the compounds and thus the method was able to quantify the compounds within the solid stool. In total, 15 compounds were quantified: Dimethyl sulphide (26–25,626 ng/g), acetone (442–3006 ng/g), ethyl butanoate (39–2468 ng/g), ethyl 2-methylbutanoate (0.3–180 ng/g), dimethyl disulphide (35–1303 ng/g), 1-octen-3-one (12 ng/g), dimethyl trisulphide (10–410 ng/g), 1-octen-3-ol (0.4–58 ng/g), ethanoic acid (672–12,963 ng/g), butanoic acid (2493–11,553 ng/g), 3-methylbutanoic acid (64–8262 ng/g), pentanoic acid (88–21,886 ng/g), indole (290–5477 ng/g), and 3-methyl indole (37–3483 ng/g). Moreover, by altering the pH of the stool to pH 13 in conjunction with the addition of 13C trimethylamine, the method was successful in detecting and quantifying trimethylamine for the first time in stool samples (range 40–5312 ng/g). Statistical analysis revealed that samples from U.K. origin had five significantly different compounds (ethyl butanoate, 1-octen-3-ol, ethanoic acid, butanoic acid, pentanoic acid, and indole) from those of South American origin. However, there were no significant differences between vegetarian and omnivore samples. These findings are supported by pre-existing literature evidence. Moreover, we have tentatively identified 12 compounds previously not reported as having been found in stool. View Full-Text
Keywords: gas-chromatography mass-spectrometry; volatile compounds; quantification; carbon isotopes; stool gas-chromatography mass-spectrometry; volatile compounds; quantification; carbon isotopes; stool
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Gould, O.; De Lacy Costello, B.; Smart, A.; Jones, P.; Macmaster, A.; Ransley, K.; Ratcliffe, N. Gas Chromatography Mass Spectrometry (GC-MS) Quantification of Metabolites in Stool Using 13C Labelled Compounds. Metabolites 2018, 8, 75.

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