Analysis of conjugated bile acids in bile by high-pressure liquid chromatography.

The major conjugated bile acids of man, including glycine and taurine conjugates, can be separated by high-pressure liquid chromatography (HPLC). The chromatographic column is 300 mm long with an internal diameter of 8.0 mm and is packed with Lichrosorb RP 18 (5 MICRONS). The mobile phase is methanol-water 75 : 25 (v/v) acidified to pH 2 with phosphoric acid. The eluent peaks are detected by a UV absorbance detector at a wavelength of 210 nm. The 10 conjugated bile acids are quantitatively analyzed on a single run of chromatography in less than 50 min. Deproteinization of biological samples is sufficient for the preparation of the analysis.