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Rapid analysis of bile acids in different biological matrices using LC-ESI-MS/MS for the investigation of bile acid transformation by mammalian gut bacteria

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Abstract

Bile acids are important signaling molecules that regulate cholesterol, glucose, and energy homoeostasis and have thus been implicated in the development of metabolic disorders. Their bioavailability is strongly modulated by the gut microbiota, which contributes to generation of complex individual-specific bile acid profiles. Hence, it is important to have accurate methods at hand for precise measurement of these important metabolites. Here, a rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous identification and quantitation of primary and secondary bile acids as well as their taurine and glycine conjugates was developed and validated. Applicability of the method was demonstrated for mammalian tissues, biofluids, and cell culture media. The analytical approach mainly consists of a simple and rapid liquid-liquid extraction procedure in presence of deuterium-labeled internal standards. Baseline separation of all isobaric bile acid species was achieved and a linear correlation over a broad concentration range was observed. The method showed acceptable accuracy and precision on intra-day (1.42–11.07 %) and inter-day (2.11–12.71 %) analyses and achieved good recovery rates for representative analytes (83.7–107.1 %). As a proof of concept, the analytical method was applied to mouse tissues and biofluids, but especially to samples from in vitro fermentations with gut bacteria of the family Coriobacteriaceae. The developed method revealed that the species Eggerthella lenta and Collinsella aerofaciens possess bile salt hydrolase activity, and for the first time that the species Enterorhabdus mucosicola is able to deconjugate and dehydrogenate primary bile acids in vitro.

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Acknowledgments

We thank Caroline Ziegler for help with bacterial cultivation. As part of the joint DFG/ANR initiative, the authors received financial support from the German Research Foundation (grant no. CL481/1-1 and RO3477/9-1) and the French National Research Agency (grant no. ANR-13-ISV3-0008-04).

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Authors and Affiliations

  1. University of Hamburg, Hamburg School of Food Science, Institute of Food Chemistry, Grindelallee 117, 20146, Hamburg, Germany

    Katrin Wegner, Laura Gau, Henrike Mueller & Sascha Rohn

  2. Junior Research Group Intestinal Microbiome, ZIEL-Institute for Food Health, Technical University of Munich, Weihenstephaner Berg 3, 85354, Freising, Germany

    Sarah Just & Thomas Clavel

  3. Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, 78350, Jouy-en-Josas, France

    Philippe Gérard & Patricia Lepage

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  1. Katrin Wegner
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Correspondence to Sascha Rohn.

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Wegner, K., Just, S., Gau, L. et al. Rapid analysis of bile acids in different biological matrices using LC-ESI-MS/MS for the investigation of bile acid transformation by mammalian gut bacteria. Anal Bioanal Chem 409, 1231–1245 (2017). https://doi.org/10.1007/s00216-016-0048-1

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