Schott, Hans -Frieder and Krautbauer, Sabrina and Hoering, Marcus and Liebisch, Gerhard and Matysik, Silke (2018) A Validated, Fast Method for Quantification of Sterols and Gut Microbiome Derived 5 alpha/beta-Stanols in Human Feces by Isotope Dilution LC-High-Resolution MS. ANALYTICAL CHEMISTRY, 90 (14). pp. 8487-8494. ISSN 0003-2700, 1520-6882
Full text not available from this repository. (Request a copy)Abstract
There has been an increasing interest during recent years in the role of the gut microbiome on health and disease. Therefore, metabolites in human feces related to microbial activity are attractive surrogate marker to track changes of microbiota induced by diet or disease. Such markers include 5 alpha/beta-stanols as microbiome-derived metabolites of sterols. Currently, reliable, robust, and fast methods to quantify fecal sterols and their related metabolites are missing. We developed a liquid chromatography-high-resolution mass spectrometry (LC-MS/HRMS) method for the quantification of sterols and their 5 alpha/beta-stanols in human fecal samples. Fecal sterols were extracted and derivatized to N, N-dimethylglycine esters. The method includes cholesterol, coprostanol, cholestanol and sitosterol, 5 alpha/beta-sitostanol, campesterol and 5 alpha/beta-campestanol. Application of a biphenyl column permits separation of isomeric 5 alpha- and 5 beta-stanols. Sterols are detected in parallel reaction monitoring (PRM) mode and stanols in full scan mode. HRMS allows differentiation of isobaric beta-stanols and the [M + 2] isotope peak of the coeluting sterol. Performance characteristics meet the criteria recommended by Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines. Analysis of fecal samples from healthy volunteers revealed high interindividual variability of sterol and stanol fractions. Interestingly, cholesterol and sitosterol showed similar fractions of mainly 5 beta-stanols. In contrast, campesterol is substantially converted to 5 alpha-campestanol and might be a poorer substrate for bacterial metabolism. Robust and fast quantification of fecal sterols and their related stanols by LC-MS/HRMS offers great potential to find novel microbiome-related biomarker in large-scale studies.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CHROMATOGRAPHY-MASS SPECTROMETRY; GAS-LIQUID-CHROMATOGRAPHY; BILE-ACIDS; NEUTRAL STEROLS; COLON-CANCER; CHOLESTEROL; EPIDEMIOLOGY; QUANTITATION; ANTIBIOTICS; CONVERSION; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 06 Mar 2020 13:01 |
| Last Modified: | 06 Mar 2020 13:01 |
| URI: | https://pred.uni-regensburg.de/id/eprint/14227 |
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