Simultaneous and rapid analysis of bile acids including conjugates by supercritical fluid chromatography coupled to tandem mass spectrometry

•A simultaneous analysis for bile acid profiling has been constructed by SFC/MS/MS.•SFC can separate 25 bile acids with a wide range of polarity.•Rat serum samples could be quantified without complicated sample preparation.

A number of analysis methods for bile acids using LC–MS and GC–MS have been reported. However, there is no reported method for the simultaneous analysis of bile acids using supercritical fluid chromatography (SFC). In this study, we have successfully developed a rapid method using SFC coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS) for comprehensive bile acid profiling. 25 bile acids including glycine and taurine conjugates were analyzed simultaneously within 13 min. The method was applied to rat serum samples, 24 of the bile acids were quantified without any solid-phase extraction and complicated sample preparation. This study not only reports simultaneous analysis of bile acids including conjugates but also indicates the method is applicable to a biological sample. This is the first report on the simultaneous analysis of bile acids using SFC/MS. The developed method will be an alternative to existing analysis methodology for studies on the bile acid metabolism in the medical and pharmaceutical fields.