Simultaneous determination of bile acids in rat liver tissue by high-performance liquid chromatography

A method for the simultaneous determination of bile acids in rat liver tissue by high-performance liquid chromatography was developed. Without prior fractionation and alkaline hydrolysis, 30 unconjugated, glycine- and taurine-conjugated bile acids were detected by post-column enzymatic reaction and fluorescence detection. They were separated on a reversed-phase column using a linear gradient solvent system of 10 mM tribasic ammonium phosphate-acetonitrile-methanol (44:12:5, v/v/v) and 20 mM dibasic ammonium phosphate-acetonitrile-methanol (2:1:2, v/v/v). The limits of detection were 1-5 pmol, and calibration curves were linear for concentrations ranging between 10 and 4000 pmol per 10 microl injection. This rapid and reliable method is effective for measuring bile acid levels in liver tissue not only of rats but also of patients with hepatobiliary and other diseases.