Analysis of bile acids by micro high-performance liquid chromatography with postcolumn enzyme reaction and fluorometric detection.

Free and conjugated bile acids in human sera were analyzed by micro high-performance liquid chromatography with detection by an immobilized hydroxysteroid dehydrogenase postcolumn reaction and a spectrofluorometer. Bile acids could be effectively collected on a precolumn, 10 X 0.2 mm I.D., packed with silica-ODS (octadecylsilane), by passing a dilute phosphate solution of a serum through it. The detection limit was 0.13-0.28 pmole for a single-to-noise ratio of 2. An amount of 0.1 ml of serum was enough for analysing each bile acid in it.