Imperial College London
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Dr. Jia Li

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Reader in Biological Chemistry
 
 
 
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Contact

 

+44 (0)20 7594 3230jia.li

 
 
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Location

 

10.N2ACommonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Gratton:2016:10.1021/acs.analchem.5b04159,
author = {Gratton, J and Phetcharaburanin, J and Mullish, BH and Williams, HRT and Thursz, M and Nicholson, JK and Holmes, E and Marchesi, JR and Li, J},
doi = {10.1021/acs.analchem.5b04159},
journal = {Analytical Chemistry},
pages = {4661--4668},
title = {An optimized sample handling strategy for metabolic profiling of human feces},
url = {http://dx.doi.org/10.1021/acs.analchem.5b04159},
volume = {88},
year = {2016}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Fecal metabolites are being increasingly studied to unravel the host-gut microbial metabolic interactions. However, there are currently no guidelines for fecal sample collection and storage based on a systematic evaluation of the effect of time, storage temperature, storage duration and sampling strategy. Here we derive an optimized protocol for fecal sample handling with the aim of maximizing metabolic stability and minimizing sample degradation. Samples obtained from five healthy individuals were analyzed to assess topographical homogeneity of feces, and to evaluate storage duration-, temperature- and freeze-thaw cycle-induced metabolic changes in crude stool and fecal water using a 1H NMR spectroscopy-based metabolic profiling approach. Inter-individual variation was much greater than that attributable to storage conditions. Individual stool samples were found to be heterogeneous and spot sampling resulted in a high degree of metabolic variation. Crude fecal samples were remarkably unstable over time and exhibited distinct metabolic profiles at different storage temperatures. Microbial fermentation was the dominant driver in time-related changes observed in fecal samples stored at room temperature and this fermentative process was reduced when stored at 4°C. Crude fecal samples frozen at -20°C manifested elevated amino acids and nicotinate and depleted short chain fatty acids compared to crude fecal control samples. The relative concentrations of branched-chain and aromatic amino acids significantly increased in the freeze-thawed crude fecal samples, suggesting a release of microbial intracellular contents. The metabolic profiles of fecal water samples were more stable compared to crude samples. Our recommendation is that intact fecal samples should be collected, kept at 4°C or on ice during transportation, and extracted ideally within 1 h of collection, or a maximum of 24 h. Fecal water samples should be extracted from a representative amount (~15 g)
AU  - Gratton,J
AU  - Phetcharaburanin,J
AU  - Mullish,BH
AU  - Williams,HRT
AU  - Thursz,M
AU  - Nicholson,JK
AU  - Holmes,E
AU  - Marchesi,JR
AU  - Li,J
DO  - 10.1021/acs.analchem.5b04159
EP  - 4668
PY  - 2016///
SN  - 0003-2700
SP  - 4661
TI  - An optimized sample handling strategy for metabolic profiling of human feces
T2  - Analytical Chemistry
UR  - http://dx.doi.org/10.1021/acs.analchem.5b04159
UR  - http://hdl.handle.net/10044/1/30855
VL  - 88
ER  -
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