Abstract
A fast and simple method was developed for the separation of unconjugated bilirubin and its mono- and di-glucuronide conjugates from bile by high-performance liquid chromatography (HPLC). Unconjugated bilirubin was separated on a reversed-phase column using acetonitrile-water (70:30 vol/vol) as the mobile phase, while the conjugates were separated on a .mu.-Bondapak-carbohydrate column employing acetonitrile-water (90:10 vol/vol) as the eluent. The application of this method was demonstrated by the analysis of the bile pigments in rat bile.