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Title: Analysis of bilirubins in biological fluids by extraction and thin-layer chromatography of the intact tetrapyrroles: application to bile of patients with Gilbert's syndrome, hemolysis, or cholelithiasis
Authors: Fevery, Johan
Blanckaert, Norbert
Leroy, P
Michiels, R
Heirwegh, K P #
Issue Date: May-1983
Series Title: Hepatology (Baltimore, Md.) vol:3 issue:2 pages:177-83
Abstract: A method was developed to extract quantitatively the bilirubins from bile, urine, serum, stool, and preparations from liver with a chloroform-ethanol mixture at pH 1.8 in the presence of ascorbic acid and NaCl. Extracted pigment was submitted to thin-layer chromatography, and the separated bilirubins were either immediately eluted and determined spectrophotometrically or individually converted to ethyl anthranilate azo derivatives for thin-layer chromatographic analysis of each isolated pigment band. Bilirubins in duodenal bile of eight healthy adults comprised 1.5 +/- 1.3% unconjugated bilirubin-IX alpha, 69 +/- 6% bilirubin diglucuronide, and 16 +/- 4% bilirubin monoglucuronides. Mixed diconjugates containing one glucuronosyl moiety and either one xylosyl or one glucosyl group amounted to 10 +/- 3%. Most samples (6 of 8) contained trace amounts (0.6 +/- 0.6%) of unconjugated bilirubin-IX beta, in agreement with nearly exclusive cleavage of heme at the alpha-meso position. The composition of the bilirubins in bile was normal in 6 patients with cholesterol gallstones, 4 with chronic hepatitis, and 3 with hemolysis. In duodenal bile of individuals with Gilbert's syndrome (n = 10), the concentration of bilirubin conjugates was comparable to that in healthy adults, but the proportion of bilirubin diglucuronides (52 +/- 8%) was decreased. The concentration of unconjugated bilirubin-IX alpha showed a fair positive correlation with that of bilirubin monoglucuronide and was increased in half of the patients with Gilbert's syndrome.
ISSN: 0270-9139
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Hepatology
Laboratory of Clinical Bacteriology and Mycology
Department of Imaging & Pathology - miscellaneous
# (joint) last author

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