Metabolite analysis of human fecal water by gas chromatography/mass spectrometry with ethyl chloroformate derivatization
Section snippets
Chemicals and materials
ECF, pyridine, and sodium hydroxide (NaOH) from Fluka (Saint Quentin en Yvelines, France) and anhydrous ethanol and n-hexane from Riedel-de Haën (Seelze, Germany) were used for derivatization reagents. l-2-Chloro-phenylalanine (a Sigma product) was used as an internal standard (IS) for batch quality control. Sodium bicarbonate and sodium sulfate were analytical reagent grade. All standards (Table 1), which were commercially obtained from Sigma–Aldrich (St. Louis, MO, USA), were prepared in
Chromatographic separation
Fecal water is a complex mixture of various metabolites, including fatty acids, amino acids, amines, and phenolic compounds, with a wide range of boiling points. The application of ECF derivatization in fecal water is beneficial not only to decrease the boiling points of metabolites but also to narrow the boiling point window of the derivatives. Consequently, capillary gas chromatography separation can cover as many compounds as possible. When the initial oven temperature was decreased from 80
Conclusions
Fecal water is a complex mixture of varieties of metabolites with a wide range of physicochemical properties and boiling points. The analytical method developed here provides a qualitative and quantitative GC/MS, with high sensitivity and efficiency, coupled with derivatization of ECF in aqueous medium. The parameters affecting derivatization efficiency and GC chromatographic separation were discussed and optimized. The pH of extraction solvent and storage conditions of fecal samples were
Acknowledgment
Xianfu Gao is thankful to INRA, France, for providing a bursary award.
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