The rate-limiting step in the overall plasma-to-bile transport of a saturating load of bilirubin is still a matter of controversy. We reassessed the apparent maximal biliary bilirubin excretion following i.v. infusion of unconjugated bilirubin and-for the first time-f highly purified bilirubin diglucuronide in the rat. The bilirubin diglucuronide preparation could be kept in a stable form at -2OOC for at least 2 months after addition of 3 mM sodium ascorbate. The biliary bilirubin excretion rates in animals with and without bile depletion in order to induce different flow rates were comparable after infusion of unconjugated bilirubin and of bilirubin diglucuronide. No significant hydrolysis of bilirubin diglucuronide seemed to occur during the hepatic transport of the pigment. Injection of bilirubin diglucuronide into rats which were already being infused with saturating doses of unconjugated bilirubin did not result in increased biliary bilirubin excretion. In contrast, a reversible inhibition of bilirubin output and bile aciddependent bile flow was observed. If unconjugated and diglucuronidated bilirubin follow the same intracellular routes, the present results would suggest that conjugation did not restrict maximal biliary excretion. However, if exogenously administered diglucuronide utilizes a separate pathway, as was recently proposed, the biliary secretion of this exogenous conjugate might be restricted, presumably due to a toxic effect of the high local concentration of diglucuronide. The pathways utilized by the unconjugated pigment, on the other hand, could be primarily determined by the conjugating capacity.
Bilirubin-IXa, the main degradation product of heme catabolism in mammals, is excreted in bile after esterification, mainly with glucuronic acid, to polar mono-and diconjugated derivatives (1-5). The overall plasma-to-