Protocol detail

Bile Acids and SCFA Quantification

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The analysis of bile acids and short chain fatty acids was performed as published previously.^{69 (link),70 (link)} Cecal content was isolated, frozen in liquid nitrogen and stored at −80°C. To avoid bacterial metabolism, further steps were performed on dry ice, including cutting for weighting. Cold Methanol was added prior homogenization and centrifugation. The Acquity UHPLC system (Waters) and a UHPLC column (Acquity™ UPLC BEH™ C8, Waters) were used for UHPLC. Mass spectrometry was performed with the amaZon ETD Ion Trap (Bruker Daltonics GmbH) in negative ionization mod and the maXis (Bruker Daltonics GmbH) in positive electorspray ionization mode for SCFAs.

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Fischer F., Romero R., Hellhund A., Linne U., Bertrams W., Pinkenburg O., Eldin H.S., Binder K., Jacob R., Walker A., Stecher B., Basic M., Luu M., Mahdavi R., Heintz-Buschart A., Visekruna A, & Steinhoff U. (2020). Dietary cellulose induces anti-inflammatory immunity and transcriptional programs via maturation of the intestinal microbiota. Gut Microbes, 12(1), 1829962.

Publication 2020

Acquity UHPLC system Acquity™ UPLC BEH™ C8 amaZon ETD Ion Trap

Bacterial Bile acids Cecal Centrifugation Cold Dry ice Frozen Mass spectrometry Metabolism Methanol Nitrogen Short chain fatty acids

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Bile acids
Short chain fatty acids

control variables

Cecal content isolation
Freezing in liquid nitrogen and storing at -80°C
Performing further steps on dry ice
Addition of cold methanol prior to homogenization and centrifugation
Use of Acquity UHPLC system and UHPLC column
Use of mass spectrometry in negative ionization mode (for bile acids) and positive electrospray ionization mode (for SCFAs)

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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