Recent Advances in the Separation and Analysis of Diazo‐Positive Bile Pigments

Heirwegh, K. P. M.; Fevery, Johan; Meuwissen, J A T P; Groote, J. De; Compernolle, Frans; Desmet, Valeer; Roy, F. P. Van

doi:10.1002/9780470110423.ch5

Cited by 113 publications

(15 citation statements)

References 163 publications

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“…To determine the presence of urobilinoids conjugated with glucuronic acid, reduction products of bilirubin diglucuronide were: (a) converted to ethylantranilate azopigments [25], that were then separated by TLC, and (b) hydrolyzed in an ammonia atmosphere and the released sugar moieties analyzed by TLC [26]. The effect of d-saccharic acid 1,4-lacone (saccharonolactone) (Sigma) (10 mM, [27]), a potent ␤-glucuronidase inhibitor, on the reduction of BDG incubated with C. perfringens was also investigated.…”

Section: Methodsmentioning

confidence: 99%

Identification of bilirubin reduction products formed by Clostridium perfringens isolated from human neonatal fecal flora

Vı́tek

Majer

Muchová

et al. 2006

Journal of Chromatography B

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Section: Methodsmentioning

confidence: 99%

Identification of bilirubin reduction products formed by Clostridium perfringens isolated from human neonatal fecal flora

Vı́tek

Majer

Muchová

et al. 2006

Journal of Chromatography B

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“…Fecal urobilinoids were determined by spectrophotometry as a zinc complexes of total urobilins [19]. Fecal bilirubin concentrations were determined by spectrophotometry as p-iodoanilin azopigments [20].…”

Section: Biochemical Analysesmentioning

confidence: 99%

The impact of intestinal microflora on serum bilirubin levels

Vı́tek

Zelenka

Zadinová

et al. 2005

Journal of Hepatology

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“…Diazo-cleavage with the diazonium salt ofp-iodoaniline or ethyl anthranilate was used to determine bile pigment concentrations (15). Thin-layer chromatographic analysis of the ethyl anthranilate azo derivatives has previously been used to estimate the relative amounts of monoconjugates, which is computed by multiplying the percentage of unconjugated azodipyrrole (an-fraction) by tlio.…”

Section: Methods Used For Comparisonmentioning

confidence: 99%

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

et al. 1983

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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‐IXα, 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‐IXβ, in agreement with nearly exclusive cleavage of heme at the α‐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 bttirubin conjugates was comparable to that in healthy adults, but the proportion of bilirubin diglucuronides (52 ± 8%) was decreased. The concentration of unconjugated bilirubin‐IXα showed a fair positive correlation with that of bilirubin monoglucuronide and was increased in half of the patients with Gilbert's syndrome.

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Recent Advances in the Separation and Analysis of Diazo‐Positive Bile Pigments

Cited by 113 publications

References 163 publications

Identification of bilirubin reduction products formed by Clostridium perfringens isolated from human neonatal fecal flora

Identification of bilirubin reduction products formed by Clostridium perfringens isolated from human neonatal fecal flora

The impact of intestinal microflora on serum bilirubin levels

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

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