Metabonomic Evaluation of Fecal Water Preparation Methods: The Effects of Ultracentrifugation
Objective: This work examines metabonomic effects of ultracentrifugation during fecal water preparation, and provides an easily adaptable method based on considerations to metabonomic impacts of the procedure.
Methods: Both fecal water (prepared from human stool samples) as well as a controlled surrogate for fecal water samples (prepared from in vitro fecal microbial communities) were used to perform systematic evaluations of various preparation steps, and used metabonomic profiles to assess the protocols’ impact on the sample.
Results: 52 metabolites were observed and found that ultracentrifugation speed decreased metabolite concentrations by approximately -2% per 10000 rpm increase, with one exception. P-cresol increased by approximately +50% per 10000 rpm. Upon investigation of the potential sources of p-cresol, we found evidence that these were tyrosine-rich cell proteins which broke down upon ultracentrifugation.
Conclusion: Based on these findings, we suggest that ultracentrifugation is an effective means of preparing fecal water samples, with negligible impact on metabonomic analyses, though measurement of p-cresol concentrations should be treated with discretion.
Document Type: Research Article
Publication date: 01 April 2018
Current Metabolomics publishes timely reviews, original research articles, thematic issues, and technical notes covering all aspects of the recent advancements and applications of metabolomics technology to systems biology, disease diagnosis, personalized medicine, and drug discovery. Original research articles and reviews in the following areas of metabolomics are of special interest to the readers of this journal:
Application to humans, plants, microbes, cell cultures, tissues, organs, and biofluids
Automation and high-throughput technology
Chemical and structural characterization of new metabolites
Chemometrics, statistical analysis, and other approaches to data analysis
Databases, including collections of reference spectra
Drug discovery, efficacy, toxicity, and personalized medicine
Integration of analytical techniques and/or various OMICS datasets
Metabolic profiling and directed metabolite analysis
New analytical experiments and techniques (NMR, MS, FTIR, etc)
Novel sensors and instrumentation for detecting metabolites
Sample preparation, extraction techniques and chromatography
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