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Free and conjugated bile pigments of body fluids: qualitative analysis by thin-layer chromatography.

Abstract The presence, in human and animal bile, of bilirubin and its mono- and diconjugates was studied by thin-layer chromatography. In health, human bile contains predominantly diconjugates; in three patients with biliary obstruction, an increased proportion of monoconjugates was observed and in one of these, this decreased after surgical treatment. The predominant pigment in bile from dog, cat, sheep, rat, pig, and ox had the mobility of a monoconjugate. Bile from the germ-free rat was identical with that from the normal rat. Only free bilirubin was detected in human gallstones. Incubation of radiolabeled bilirubin diconjugate with β-glucuronidase demonstrated sequential liberation of bilirubin monoglucuronide and free bilirubin.