Extraction and quantitative determination of bile acids in feces
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 metho...
| Main Authors: | , , , , , |
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| Format: | Journal Article |
| Language: | English |
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ELSEVIER
2021
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.11937/86513 |
| _version_ | 1848764842241949696 |
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| author | Shafaei, A. Rees, J. Christophersen, Claus Devine, A. Broadhurst, D. Boyce, M.C. |
| author_facet | Shafaei, A. Rees, J. Christophersen, Claus Devine, A. Broadhurst, D. Boyce, M.C. |
| author_sort | Shafaei, A. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T11:25:47Z |
| format | Journal Article |
| id | curtin-20.500.11937-86513 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T11:25:47Z |
| publishDate | 2021 |
| publisher | ELSEVIER |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-865132021-11-29T06:42:44Z Extraction and quantitative determination of bile acids in feces Shafaei, A. Rees, J. Christophersen, Claus Devine, A. Broadhurst, D. Boyce, M.C. Science & Technology Physical Sciences Chemistry, Analytical Chemistry Bile acids Fecal LC-MS MS Extraction Stability 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. 2021 Journal Article http://hdl.handle.net/20.500.11937/86513 10.1016/j.aca.2021.338224 English ELSEVIER restricted |
| spellingShingle | Science & Technology Physical Sciences Chemistry, Analytical Chemistry Bile acids Fecal LC-MS MS Extraction Stability Shafaei, A. Rees, J. Christophersen, Claus Devine, A. Broadhurst, D. Boyce, M.C. Extraction and quantitative determination of bile acids in feces |
| title | Extraction and quantitative determination of bile acids in feces |
| title_full | Extraction and quantitative determination of bile acids in feces |
| title_fullStr | Extraction and quantitative determination of bile acids in feces |
| title_full_unstemmed | Extraction and quantitative determination of bile acids in feces |
| title_short | Extraction and quantitative determination of bile acids in feces |
| title_sort | extraction and quantitative determination of bile acids in feces |
| topic | Science & Technology Physical Sciences Chemistry, Analytical Chemistry Bile acids Fecal LC-MS MS Extraction Stability |
| url | http://hdl.handle.net/20.500.11937/86513 |