TY - JOUR

T1 - High-Throughput UHPLC-MS to Screen Metabolites in Feces for Gut Metabolic Health

AU - de Zawadzki, Andressa

AU - Thiele, Maja

AU - Suvitaival, Tommi

AU - Wretlind, Asger

AU - Kim, Min

AU - Ali, Mina

AU - Bjerre, Annette F.

AU - Stahr, Karin

AU - Mattila, Ismo

AU - Hansen, Torben

AU - Krag, Aleksander

AU - Legido-Quigley, Cristina

N1 - Publisher Copyright: © 2022 by the authors.Licensee MDPI, Basel, Switzerland

PY - 2022

Y1 - 2022

N2 - 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. 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 nonpolar 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. 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 metabolitewise relative standard deviation (RSD) in quality control samples, inter-day repeatability, LOD, LOQ, range of linearity and method recovery. The average concentrations for 135 healthy participants are reported here for clinical applications. Our high-throughput method provides a novel tool for investigating gut-liver axis metabolism in liver-related diseases using a noninvasive collected sample.

AB - 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. 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 nonpolar 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. 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 metabolitewise relative standard deviation (RSD) in quality control samples, inter-day repeatability, LOD, LOQ, range of linearity and method recovery. The average concentrations for 135 healthy participants are reported here for clinical applications. Our high-throughput method provides a novel tool for investigating gut-liver axis metabolism in liver-related diseases using a noninvasive collected sample.

KW - Bile acids

KW - Fecal metabolomics

KW - Gut-liver axis

KW - Sample preparation

KW - Targeted metabolomics

U2 - 10.3390/metabo12030211

DO - 10.3390/metabo12030211

M3 - Journal article

C2 - 35323654

AN - SCOPUS:85128521896

VL - 12

JO - Metabolites

JF - Metabolites

SN - 2218-1989

IS - 3

M1 - 211

ER -