RT Journal Article
SR Electronic
T1 High-throughput UHPLC-MS to screen metabolites in feces for gut metabolic health
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.12.22.473790
DO 10.1101/2021.12.22.473790
A1 Andressa de Zawadzki
A1 Maja Thiele
A1 Tommi Suvitaival
A1 Asger Wretlind
A1 Min Kim
A1 Mina Ali
A1 Annette F. Bjerre
A1 Karin Stahr
A1 Ismo Matilla
A1 Torben Hansen
A1 Aleksander Krag
A1 Cristina Legido-Quigley
YR 2021
UL http://biorxiv.org/content/early/2021/12/23/2021.12.22.473790.abstract
AB Background Feces are the product of our diets and have been linked to diseases of the gut, including Chron’s disease and metabolic diseases such as diabetes. For screening metabolites in heterogeneous samples such as feces, it is necessary to use fast and reproducible analytical methods that maximize metabolite detection.Methods As sample preparation is crucial to obtain high quality data in MS-based clinical metabolomics, we developed a novel, efficient and robust method for preparing fecal samples for analysis with a focus in reducing aliquoting and detecting both polar and non-polar metabolites. Fecal samples (n= 475) from patients with alcohol-related liver disease and healthy controls were prepared according to the proposed method and analyzed in an UHPLC-QQQ targeted platform in order to obtain a quantitative profile of compounds that impact liver-gut axis metabolism.Results MS analyses of the prepared fecal samples have shown reproducibility and coverage of n=28 metabolites, mostly comprising bile acids and amino acids. We report metabolite-wise relative standard deviation (RSD) in quality control samples, inter-day repeatability, LOD, LOQ and range of linearity. The average concen-trations for 135 healthy participants are reported here for clinical applications.Conclusions our high-throughput method provides an efficient tool for investigating gut-liver axis metabolism in liver-related diseases using a noninvasive collected sample.Competing Interest StatementThe authors have declared no competing interest.