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Optimized Sample Handling Strategy for Metabolic Profiling of Human Feces
Jasmine Gratton, Jutarop Phetcharaburanin, Benjamin H Mullish
1Center for Digestive and Gut Health, Institute of Global Health Innovation, Imperial College London , South Kensington, London SW7 2AZ, U.K.
Analytical Chemistry
|April 12, 2016
Summary
Optimized fecal sample handling is crucial for studying host-gut microbial interactions. This study recommends collecting intact fecal samples, storing them at 4°C, and processing within 24 hours to ensure metabolic stability.
Area of Science:
- Microbiology
- Metabolomics
- Analytical Chemistry
Background:
- Host-gut microbial metabolic interactions are vital for health.
- Lack of standardized fecal sample collection and storage protocols hinders research.
- Systematic evaluation is needed to optimize fecal sample handling for metabolic stability.
Purpose of the Study:
- To develop an optimized protocol for fecal sample collection and storage.
- To assess the impact of time, temperature, and freeze-thaw cycles on fecal metabolic profiles.
- To minimize metabolic degradation and maximize stability in fecal samples for analysis.
Main Methods:
- Metabolic profiling of fecal samples using (1)H NMR spectroscopy.
- Analysis of topographical homogeneity within individual stool samples.
- Evaluation of storage duration, temperature, and freeze-thaw effects on crude stool and fecal water.
Main Results:
- Interindividual variation in fecal metabolites significantly exceeded variations due to storage conditions.
- Crude fecal samples showed instability over time and temperature variations, with microbial fermentation being a key factor at room temperature.
- Fecal water samples exhibited greater metabolic stability compared to crude samples, with specific changes noted in amino acids and fatty acids upon freezing and thawing.
Conclusions:
- Intact fecal samples should be collected, transported at 4°C or on ice, and processed within 24 hours.
- Fecal water extraction requires homogenization of approximately 15g of stool, followed by aliquoting and storage below -20°C, avoiding freeze-thaw cycles.
- Standardized fecal sample handling protocols are essential for reliable host-gut microbial metabolic interaction studies.
