Abstract
A rapid and sensitive method has been developed for the simultaneous determination of free bile acids and its glyco-and tauro-conjugates by high-performance liquid chromatography (HPLC). These components were separated on a resin-based reversed-phase column, with a gradient of acetonitrile in dilute sodium hydroxide solution. The detection was performed at a gold electrode with a sequence of three pulses, which enabled the selective detection of hydroxyl-containing compounds, without fouling of the electrode. Detection limits ranged from 30 to 60 pmol oncolumn. Linear calibration graphs were obtained for bile acid solutions of 0.6 to 14 μmol/l. The method was applied to the determination of bile acids in culture media, bile, and ileal fluid.
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