BILE PIGMENT ANALYSIS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Abstract

A new high performance liquid chromatographic (HPLC) method was developed for analysis of bile pigments using a reversed phase partition method. This method allows a better separation of a variety of conjugated and unconjugated bilirubin than similar HPLC methods previously reported, and the xyloside conjugate which was not detected by other HPLC methods was identified. There is no degradation of bilirubin tetrapyrroles during the total 90 minute period for this HPLC procedure. The bile of healthy adult subjects was found to contain bilirubin diglucuronide (75.4%), bilirubin monoglucuronide (10.4%), bilirubin monoglucuronide monoglucoside diester (7.4%), bilirubin monoglucuronide monoxyloside diester (3.0%), unidentified bilirubin tetrapyrrole (2.7%) and bilirubin IX_α (1.1%).