Abstract
There is a growing interest in the named “acidic sterolbiome” and in the genetic potential of the gut microbiome (GM) to modify bile acid (BA) structure. Indeed, the qualitative composition of BAs in feces correlates with the bowel microorganisms and their collective genetic material. GM is responsible for the production of BA metabolites, such as secondary and oxo-BAs. The specific BA profiles, as microbiome-host co-metabolic products, could be useful to investigate the GM-host interaction in animals under physiological conditions, as well as in specific diseases. In this context, we developed and validated an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry method for the simultaneous analysis of up to 21 oxo-BAs and their 9 metabolic precursors. Chromatographic separation was achieved in 7 min with adequate analytical performance in terms of selectivity, sensitivity (LOQ from 0.05 to 0.1 µg/mL), accuracy (bias% < 5%), precision (CV% < 5%) and matrix effect (ME% < 10%). A fast solvent extraction protocol has been fine-tuned, achieving recoveries > 90%. In parallel, the gut microbiota assessment in farming animals was evaluated by 16S rRNA next-generation sequencing, and the correlation with the BA composition was performed by multivariate analysis, allowing to reconstruct species-specific associations between the BA profile and specific GM components.
Highlights
There is a growing interest in the named “acidic sterolbiome” and in the genetic potential of the gut microbiome (GM) to modify bile acid (BA) structure
BAs are recycled in the enterohepatic circulation, where they exert their physiological function in bile to facilitate lipid absorption via mixed micelles thanks to their detergency
No other class of small molecules in vertebrates exhibits the variety of chemical structures shown by BAs, and the identification of new bile alcohols and BA derivatives continues to date[3]
Summary
There is a growing interest in the named “acidic sterolbiome” and in the genetic potential of the gut microbiome (GM) to modify bile acid (BA) structure. The specific BA profiles, as microbiome-host co-metabolic products, could be useful to investigate the GM-host interaction in animals under physiological conditions, as well as in specific diseases In this context, we developed and validated an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry method for the simultaneous analysis of up to 21 oxo-BAs and their 9 metabolic precursors. As already demonstrated, they are ligands of nuclear receptors, such as farnesoid X receptor (FXR), largely expressed in the liver and intestine, which is involved in several biological pathways, including BA own synthesis The finding of their “hormone activity” at the end of the twentieth century has elicited an increasing interest in the BAs family, especially for their potential use as drugs in several hepato-biliary diseases. The study of BA variations across vertebrates, has highlighted a clear evidence of the evolutionary transition from C27 ancestral bile alcohols to C24 BAs, with an intermediate phase in some animal classes where C27 BAs are present
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