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    Extraction and quantitative determination of bile acids in feces

    Access Status
    Fulltext not available
    Authors
    Shafaei, A.
    Rees, J.
    Christophersen, Claus
    Devine, A.
    Broadhurst, D.
    Boyce, M.C.
    Date
    2021
    Type
    Journal Article
    
    Metadata
    Show full item record
    Citation
    Shafaei, A. and Rees, J. and Christophersen, C.T. and Devine, A. and Broadhurst, D. and Boyce, M.C. 2021. Extraction and quantitative determination of bile acids in feces. Analytica Chimica Acta. 1150: Article No. 338224.
    Source Title
    Analytica Chimica Acta
    DOI
    10.1016/j.aca.2021.338224
    ISSN
    0003-2670
    Faculty
    Faculty of Science and Engineering
    School
    School of Molecular and Life Sciences (MLS)
    URI
    http://hdl.handle.net/20.500.11937/86513
    Collection
    • Curtin Research Publications
    Abstract

    With rapid advances in gut microbiome research, fecal bile acids are increasingly being monitored as potential biomarkers of diet related disease susceptibility. As such, rapid, robust and reliable methods for their analysis are of increasing importance. Herein is described a simple extraction method for the analysis of bile acids in feces suitable for subsequent quantification by liquid chromatography and tandem mass spectrometry. A C18 column separated the analytes with excellent peak shape and retention time repeatability maintained across 800 injections. The intra-day and inter-day precision and accuracy was greater than 80%. Recoveries ranged from 83.58 to 122.41%. The limit of detection and limit of quantification were in the range 2.5–15 nM, respectively. The optimized method involved extracting bile acids from wet feces with minimal clean up. A second aliquot of fecal material was dried and weighed to correct for water content. Extracting from dried feces showed reduced recovery that could be corrected for by spiking the feces with deuterated standards prior to drying. Storage of the extracts and standards in a refrigerated autosampler prior to analysis on the LC-MS is necessary. Multiple freeze-thaws of both extracts and standards lead to poor recoveries for some bile acids. The method was successfully applied to 100 human fecal samples.

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