Quantitative liquid-chromatographic estimation of bilirubin species in pathological serum.

Lauff, John J.; Kasper, Mary E.; Ambrose, Robert T.

doi:10.1093/clinchem/29.5.800

Cited by 47 publications

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“…Thi s suggests that the pattern of bilirubin conjugation in infants may favor monoester over diester conjugation: a one-step conjug ation process that appears to be more efficient because only one sugar molecule is used for conjugation to each bilirubin molecule (13). However, because interconver sion of diester derivatives of bilirubin to mono esters can also occur by spontaneou s rearrangement and hydrolysis (8), the clinical significance of the disproportion between monoand diconjugates , as noted above, is stilI not clear; (c) the routinely used diazo method for bilirubin analysis underestimated the direct bilirubin fraction by as much as 34% compared with HPLC, a finding similar to what has been observed in adult and pediatric patients (4,14). Admittedly, there are some inherent problems involved in the quantitation of bilirubin species by an y method .…”

Section: Resultsmentioning

confidence: 66%

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The Occurrence and Significance of the Bilirubin Species, Including Delta Bilirubin, in Jaundiced Infants

Ostrea,

Ongtengco,

Tolia

et al. 1988

J. pediatr. gastroenterol. nutr.

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Summary:An improved technique for bilirubin analysis, using high‐performance liquid chromatography, has enabled us to study the occurrence and significance of four species of bilirubin (unconjugated, monoconjugated, diconjugated, and delta bilirubin) in 40 infants with jaundice of various etiologies. We found that: (a) infants with indirect hyperbilirubinemia showed >90% of their total serum bilirubin as unconjugated bilirubin. The small remaining fraction consisted of conjugated bilirubin; predominantly delta bilirubin (5%); (b) infants with elevated direct serum bilirubin (≥2 mg/dl) showed almost twice more monoconjugated than diconjugated bilirubin fractions; (c) the standard diazo test for bilirubin analysisunderestimates the direct bilirubin by as much as 34%; and (d) delta bilirubin, a tightly protein bound bilirubin, was observed in significant amounts in infants with elevateddirect bilirubin. Its concentration, which ranged from 10–73% of the total bilirubin, was related to the duration rather than to the cause of the jaundice. It was also observed at birth in an infant with giant‐cell hepatitis. It is concluded that the identification of more specific bilirubin species in jaundiced infants, especially in those with elevated direct serum bilirubin, will further help in the understanding and management of their disease.

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Section: Resultsmentioning

confidence: 66%

“…Recently, new techniques in high-performance liquid ch romatography (H PLC) have resulted in the identification of another type of bilirubin , referred to as delt a bilirubin or biliprot ein. Thi s bilirubin reacts directly with the diazo reagent and is characteristica lly tightl y (covalently) bound to albumin (4,5).…”

mentioning

confidence: 99%

The Occurrence and Significance of the Bilirubin Species, Including Delta Bilirubin, in Jaundiced Infants

Ostrea,

Ongtengco,

Tolia

et al. 1988

J. pediatr. gastroenterol. nutr.

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“…Das Gesamtbilirubin des Serums setzt sich aus 4 verschiedenen Fraktionen zusammen: dem unkonjugierten -Bilirubin, das über hydrophobe Wechselwirkungen an Albumin gebunden ist, dem mono-und diglucuronidierten ß-bzw. -Bilirubin, das in gelöster Form vorliegt sowie einer kovalent an Albumin gebundenen 8-Fraktion (5,6). Die klassische Bilirubinbestimmungsmethode von Jendrassik & Grof(\) unterscheidet allerdings nur 2 Fraktionen, das hydrophilere "direkte" Bilirubin ( , , ), das im wäßrigen Milieu rasch reagiert und das hydrophobere "indirekte" Bilirubin, das üblicherweise erst durch Zusatz von Lösungsvermittlern erfaßt wird (l, 7).…”

Section: Diskussionunclassified

Reaktion von direktem und indirektem Bilirubin mit Fast Red TR und verwandten Diazoniumsalzen

Hoffmann¹,

Hiefinger²,

Weiss³

1985

Clinical Chemistry and Laboratory Medicine

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Zusammenfassung:In der vorliegenden Arbeit wird die Reaktivität von 15 verschiedenen Diazoniumsalzen gegenüber Bilirubin im Serum untersucht. Es wird gezeigt, daß die Selektivität der Bestimmung von direktem und gesamtem Bilirubin vor allem von zwei Bedingungen abhängig ist: vom pH-Wert der Reaktionslösung und von der Art und Stellung der Substituenten am Benzolring. Durch Einführung "elektronenschiebender" Gruppen wie z. B. der -Methylgruppe im Fast Red TR läßt sich die Reaktivität des Diazoniumkations gegenüber Bilirubin gezielt so verändern, daß bei pH-Werten zwischen 2 und 5 die Mitreaktion der indirekten Fraktion auf ein Minimum reduziert ist. Nach Neutralisierung auf pH 7 läßt sich das gesamte Bilirubin bestimmen. Reaction of non^conjugated and conjugated bilirubin with Fast Red TR and related diazonium salts

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“…in 1981 [3,4]. Its clinical significance has been the subject of several subsequent studies [5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Analyses of azopigments obtained from the delta fraction of bilirubin from mammalian plasma (mammalian biliprotein)

Yoshida

Inagaki

Hirano

et al. 1987

Biochemical Journal

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Azopigments were obtained from the delta fraction of bilirubin (mammalian biliprotein) in cholestatic sera of men, rats and guinea pigs by diazo reaction with diazotized p-iodoaniline and analysed by t.l.c. Delta bilirubin of men and rats generated both unconjugated and glucuronide-conjugated azodipyrroles, whereas that of guinea pigs, in which the predominant form of conjugated bilirubin in serum was bilirubin monoglucuronide, generated only unconjugated azodipyrrole. We further analysed the azopigments by reversed-phase h.p.l.c. to distinguish their endovinyl and exovinyl isomers. The results indicated (a) that covalent binding of bilirubin to protein occurs exclusively on the conjugated dipyrrolic (either endovinyl or exovinyl) half of the parent conjugated bilirubin, (b) that both bilirubin monoglucuronide and bilirubin diglucuronide generate delta bilirubin, the latter yielding a 'conjugated' form of delta bilirubin that preserves the glucuronic acid moiety on the dipyrrolic half not bound covalently to protein, and (c) that therefore at least four forms of delta bilirubin exist in jaundiced sera of men and rats.

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Quantitative liquid-chromatographic estimation of bilirubin species in pathological serum.

Cited by 47 publications

References 0 publications

The Occurrence and Significance of the Bilirubin Species, Including Delta Bilirubin, in Jaundiced Infants

The Occurrence and Significance of the Bilirubin Species, Including Delta Bilirubin, in Jaundiced Infants

Reaktion von direktem und indirektem Bilirubin mit Fast Red TR und verwandten Diazoniumsalzen

Analyses of azopigments obtained from the delta fraction of bilirubin from mammalian plasma (mammalian biliprotein)

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