Abstract
Fecal metabolomics-based analysis indisputably constitutes a very useful tool for elucidating the biochemistry of digestion and absorption of the gastrointestinal system. Fecal samples represent the most suitable, non-invasive, specimen for the study of the symbiotic relationship between the host and the intestinal microbiota.
It is well established that the balance of the intestinal microbiota changes in response to some stimuli, physiological such as gender, age, diet, exercise and pathological such as gastrointestinal and hepatic disease. Fecal samples have been analyzed using the most widespread analytical techniques, namely, NMR spectroscopy, GC-MS, and LC-MS/MS. Rat fecal sample is a frequently used and particularly useful substrate for metabolomics-based studies in related fields. The complexity and diversity of the nature of fecal samples require careful and skillful handling for the effective quantitative extraction of the metabolites while avoiding their deterioration. Parameters such as the fecal sample weight to extraction solvent volume, the nature and the pH value of the extraction solvent, and the homogenization process are some important factors for the optimal extraction of samples, in order to obtain high-quality metabolic fingerprints, using either untargeted or targeted metabolomics.
Key words
- Metabolomics
- Sample preparation
- Fecal samples
- Rats
- NMR
- GC-MS
- LC-MS/MS
- Fecal extract
This is a preview of subscription content, access via your institution.
Buying options
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Guinane CM, Cotter PD (2013) Role of the gut microbiota in health and chronic gastrointestinal disease: understanding a hidden metabolic organ. Ther Adv Gastroenterol 6(4):295–308
Holmes E, Li JV, Athanasiou T, Ashrafian H, Nicholson JK (2011) Understanding the role of gut microbiome-host metabolic signal disruption in health and disease. Trends Microbiol 19(7):349–359
Deda O, Gika HG, Wilson ID, Theodoridis GA (2015) An overview of fecal sample preparation for global metabolic profiling. J Pharm Biomed Anal 113:137–150
Theodoridis G, Gika H, Franceschi P et al (2012) LC-MS based global metabolite profiling of grapes: solvent extraction protocol optimisation. Metabolomics 8(2):175–185
Gika H, Theodoridis G (2011) Sample preparation prior to the LC-MS-based metabolomics/metabonomics of blood-derived samples. Bioanalysis 3(14):1647–1661
Gika HG, Wilson ID, Theodoridis GA (2014) LC–MS-based holistic metabolic profiling. Problems, limitations, advantages, and future perspectives. J Chromatogr B 966:1–6
Bollard ME, Stanley EG, Lindon JC et al (2005) NMR-based metabonomic approaches for evaluating physiological influences on biofluid composition. NMR Biomed 18(3):143–162
Deda O, Chatziioannou AC, Fasoula S et al (2017) Sample preparation optimization in fecal metabolic profiling. J Chromatogr B 1047:115–123
Hooper LV, Midtvedt T, Gordon JI (2002) How host-microbial interactions shape the nutrient environment of the mammalian intestine. Annu Rev Nutr 22:283–307
Lamichhane S, Yde CC, Schmedes MS et al (2015) Strategy for nuclear-magnetic-resonance-based metabolomics of human feces. Anal Chem 87(12):5930–5937
Wu J, An Y, Yao J, Wang Y, Tang H (2010) An optimised sample preparation method for NMR-based faecal metabonomic analysis. Analyst 135(5):1023–1030
Saric J, Wang Y, Li J et al (2008) Species variation in the fecal metabolome gives insight into differential gastrointestinal function. J Proteome Res 7(1):352–360
Deda O, Gika H, Panagoulis T et al (2017) Impact of exercise on fecal and cecal metabolome over aging: a longitudinal study in rats. Bioanalysis 9(1):21–36
Bezabeh T, Somorjai RL, Smith IC (2009) ICP MR metabolomics of fecal extracts: applications in the study of bowel diseases. Magn Reson Chem 47(S1):S54–S61
Monleon D, Garcia-Valles R, Morales JM et al (2014) Metabolomic analysis of long-term spontaneous exercise in mice suggests increased lipolysis and altered glucose metabolism when animals are at rest. J Appl Physiol 117(10):1110–1119
Gao X, Pujos-Guillot E, Martin J-F et al (2009) Metabolite analysis of human fecal water by gas chromatography/mass spectrometry with ethyl chloroformate derivatization. Anal Biochem 393(2):163–175
Gao X, Pujos-Guillot E, Sébédio J-L (2010) Development of a quantitative metabolomic approach to study clinical human fecal water metabolome based on trimethylsilylation derivatization and GC/MS analysis. Anal Chem 82(15):6447–6456
Virgiliou C, Sampsonidis I, Gika HG, Raikos N, Theodoridis GA (2015) Development and validation of a HILIC-MS/MS multitargeted method for metabolomics applications. Electrophoresis 36(18):2215–2225
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Deda, O., Gika, H.G., Theodoridis, G.A. (2018). Rat Fecal Metabolomics-Based Analysis. In: Theodoridis, G., Gika, H., Wilson, I. (eds) Metabolic Profiling. Methods in Molecular Biology, vol 1738. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7643-0_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7643-0_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7642-3
Online ISBN: 978-1-4939-7643-0
eBook Packages: Springer Protocols