A fast and simple ion-pair high performance liquid chromatography method for analysis of primary bile salts in in vitro digested bean samples

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Bile salts (BS) play a key role in cholesterol and lipid metabolism as well as in many other key metabolic pathways. High performance liquid chromatography (HPLC) is the most common technique used to analyze BS in diverse type of samples. However, current HPLC analysis methods used to analyze and quantify single BS in in vitro digested samples showed poor separation of a complex mixture of BS. In this article, we improved a standard method originally used for quantifying individual BS in food samples subjected to in vitro digestion. We also adapted a method previously developed for BS examination in human blood samples to the analysis of these molecules in chyme samples obtained during simulated gastrointestinal digestion. Our method was simple and achieved a fast and successful separation and quantification of four primary BS (sodium salts of taurocholic, glycocholic, taurochenodeoxycholic and glycochenodeoxycholic acids). •A method used to analyze bile salts in human blood samples has been adapted to separate and quantify four primary bile salts in in vitro digested bean samples. •Addition of an ion-pair reagent led to complete separation of glycine and taurine conjugates of chenodeoxycholic and cholic acids within 10 min, and achieved good peak symmetry. •The minimum BS concentration that could be measured was as low as 0.03125mM.



Bile salts, HPLC, Ion-pair reagent, In vitro digestion, Chyme samples