Disturbances of bilirubin metabolism such as jaundice or pigment gallstone formation occur during total parenteral nutrition (TPN). We have studied the effects of TPN on bile flow and bile acid secretion and on the hepatobiliary transport of bilirubin in rats. Animals on parenteral nutrition for 5 days received 4.8 g of amino acids and 6.9 g of glucose daily. Controls were orally fed animals. Bile flow and bile acid secretion were not significantly modified by TPN. Serum bile acid and alkaline phosphatase levels were significantly increased in TPN animals when compared with the controls (+98% and +38%, respectively), which points to a relative cholestasis in the TPN rats. The biliary excretion of bilirubin monoconjugates and bilirubin diconjugates was significantly increased (+72% and +78%, respectively). This provides evidence for enhanced production of the pigment. Serum concentration of total bilirubin was enhanced in the TPN rats (+240%). The esterified/total bilirubin ratio in serum increased, whereas the bilirubin diconjugates/bilirubin monoconjugates ratio decreased. These facts, together with the minor reduction of hepatic bilirubin UDP glucuronosyltransferase activity (-12%), suggest that hyperbilirubinemia would be a consequence of both cholestasis and increased bilirubin production. The alterations reported here could contribute to the explanation of hyperbilirubinemia and pigment gallstone formation in patients maintained on parenteral nutrition.