High pressure liquid chromatography solvent systems for studies of bile acid biosynthesis

Reversed phase high pressure liquid chromatography (HPLC) solvent systems have been developed for the separation of intermediates in the formation of bile acids and bile acid conjugates from cholesterol. Four different mobile phases (water-methanol, 10 mM acetate buffer (pH 4.37)-methanol, 30 mM trifluoroacetic acid (pH 2.9 with triethylamine)-methanol, and 50 mM potassium phosphate buffer (pH 7.0)-2-propanol) have been applied to obtain separation of all the main intermediates with use of the same reversed phase column (Zorbax ODS).