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. 1977 Oct 1;167(1):1–8. doi: 10.1042/bj1670001

The isolation and further characterization of the bilirubin tetrapyrroles in bile-containing human duodenal juice and dog gall-bladder bile

Ellen R Gordon 1,*, Tak-Hang Chan 1, Karen Samodai 1, Carl A Goresky 1
PMCID: PMC1183615  PMID: 588243

Abstract

Bilirubin and its conjugates were extracted from either dog gall-bladder bile or bile-containing human duodenal juice into chloroform containing 10mm-tetraheptylammonium chloride. The intact bilirubin tetrapyrroles were then separated by t.l.c. Structural elucidation was made after coupling of the individual pigments with diazonium salts. Four azopigments were detected: azopigment αo or dipyrrolic azobilirubin; azopigment δ or dipyrrolic azobilirubin monoglucuronide; azopigment α3 or dipyrrolic azobilirubin monoglucoside; and, from dog gall-bladder bile, azopigment α2. The last conjugate required further verification of its structure. After methanolysis, it was shown by combined g.l.c.–mass spectrometry to contain xylose in a 1:1 molar ratio with the azopigments of bilirubin. Human bile contained 86% bilirubin diglucuronide, 7% bilirubin monoglucuronide monoglucoside diester, 4% bilirubin monoglucuronide and 3% bilirubin. Dog gall-bladder bile had a considerably different composition; it contained 47% bilirubin diglucuronide, 40% bilirubin monoglucuronide monoglucoside diester, 8% bilirubin monoglucuronide, 4% bilirubin diglucoside, 1–2% bilirubin and traces of conjugates containing xylose. The total bilirubin content and proportions of the conjugates did not change in bile that was frozen and stored at −20°C under N2, whereas in the chloroform/tetraheptylammonium chloride extract, similarly stored, total pigment was slowly lost and the diglucuronide conjugate converted into the monoglucuronide.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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