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@ARTICLE{Erben:289354,
      author       = {V. Erben$^*$ and G. Poschet and P. Schrotz-King$^*$ and H.
                      Brenner$^*$},
      title        = {{E}valuation of different stool extraction methods for
                      metabolomics measurements in human faecal samples.},
      journal      = {BMJ nutrition, prevention $\&$ health},
      volume       = {4},
      number       = {2},
      issn         = {2516-5542},
      address      = {London},
      publisher    = {BMJ Publishing Group Limited},
      reportid     = {DKFZ-2024-00744},
      pages        = {374 - 384},
      year         = {2021},
      note         = {#EA:C120#LA:C120#LA:C070#},
      abstract     = {Metabolomics analysis of human stool samples is of great
                      interest for a broad range of applications in biomedical
                      research including early detection of colorectal neoplasms.
                      However, due to the complexity of metabolites there is no
                      consensus on how to process samples for stool metabolomics
                      measurements to obtain a broad coverage of hydrophilic and
                      hydrophobic substances.We used frozen stool samples (50 mg)
                      from healthy study participants. Stool samples were
                      processed after thawing using eight different processing
                      protocols and different solvents (solvents such as
                      phosphate-buffered saline, isopropanol, methanol, ethanol,
                      acetonitrile and solvent mixtures with or without following
                      evaporation and concentration steps). Metabolites were
                      measured afterwards using the MxP Quant 500 kit (Biocrates).
                      The best performing protocol was subsequently applied to
                      compare stool samples of participants with different dietary
                      habits.In this study, we were able to determine up to 340
                      metabolites of various chemical classes extracted from stool
                      samples of healthy study participants with eight different
                      protocols. Polar metabolites such as amino acids could be
                      measured with each method while other metabolite classes,
                      particular lipid species (better with isopropanol and
                      ethanol or methanol following a drying step), are more
                      dependent on the solvent or combination of solvents used.
                      Only a small number of triglycerides or acylcarnitines were
                      detected in human faeces. Extraction efficiency was higher
                      for protocols using isopropanol (131 metabolites>limit of
                      detection (LOD)) or those using ethanol or methanol and
                      methyl tert-butyl ether (MTBE) including an evaporation and
                      concentration step (303 and 342 metabolites>LOD,
                      respectively) than for other protocols. We detected
                      significant faecal metabolite differences between
                      vegetarians, semivegetarians and non-vegetarians.For the
                      evaluation of metabolites in faecal samples, we found
                      protocols using solvents like isopropanol and those using
                      ethanol or methanol, and MTBE including an evaporation and
                      concentration step to be superior regarding the number of
                      detected metabolites of different chemical classes over
                      others tested in this study.},
      keywords     = {dietary patterns (Other)},
      cin          = {C070 / HD01 / C120},
      ddc          = {610},
      cid          = {I:(DE-He78)C070-20160331 / I:(DE-He78)HD01-20160331 /
                      I:(DE-He78)C120-20160331},
      pnm          = {313 - Krebsrisikofaktoren und Prävention (POF4-313)},
      pid          = {G:(DE-HGF)POF4-313},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:35028509},
      pmc          = {pmc:PMC8718864},
      doi          = {10.1136/bmjnph-2020-000202},
      url          = {https://inrepo02.dkfz.de/record/289354},
}