Analysis of bilirubin conjugates in human bile by column liquid chromatography—Changes in their composition in hepatobiliary diseases

Bilirubin in human bile was analysed by high performance liquid chromatography (HPLC), and seven peaks were identifiable. There was no difference in the proportion of bilirubin between the B bile and C bile from healthy humans obtained by the Meltzer‐Lyon method. HPLC coupled with nuclear magnetic resonance spectroscopy on bilirubin after diazotization permitted differentiation between endovinyl and exovinyl conjugates of bilirubin. In patients with Gilbert's syndrome, type II Crigler‐Najjar syndrome, and haemolytic anaemia, the proportion of bilirubin diglucuronide (BDG) in the C bile decreased, and that of bilirubin monoglucuronide (BMG) increased. In patients with type II Crigler‐Najjar syndrome, marked elevations in BMG and bilirubin IXα and decreases in bilirubin monoglucuronide monoglucoside diester (BGG) and bilirubin monoglucuronide monoxyloside diester (BGX) were observed. In normal subjects and patients with unconjugated hyperbilirubinaemia, the diazobased indirect‐reacting bilirubin levels in blood and the proportion of BMG in bile were in good correlation. In patients with acute hepatitis, chronic hepatitis, and liver cirrhosis, there was no difference in the bilirubin composition in bile as compared to normal subjects, except for slight elevations in BGG observed in patients with chronic hepatitis and liver cirrhosis. In gallbladder bile obtained from patients with cholelithiasis, bilirubin IXα increased, suggesting some deconjugation of BDG.