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PubMed · 2773134

[Fractionated P-bilirubins].

C S Schou·H Mortensen
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Abstract

A diazo-based dry film technique for the estimation of different bilirubins in plasma is now available. This procedure separates bilirubins from icteric sera into three separate fractions: bilirubin (unconjugated), bilirubin-glucuronides (mono + diglucuronide) and bilirubin-albumin. In newborns with prolonged jaundice classification of hyperbilirubinemia is of importance for choice of treatment. While binding of bilirubin and bilirubin-glucuronides to albumin is non covalent, reversible, bilirubin-albumin appears to be firmly associated with albumin by covalent bonds. This causes delayed clearance of this bilirubin fraction from plasma as the half-life of albumin is approximately 18 days. Hence the substance concentration of bilirubin-albumin will decrease at a slower rate than will bilirubin and bilirubin-glucuronide, despite hepatobiliary recovery. Bilirubin-albumin may therefore prove of value in the differentiation between different clinical entities with hyperbilirubinemia.

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BilirubinHumansHyperbilirubinemiaInfant, NewbornJaundice, NeonatalSerum AlbuminEnglish AbstractJournal Article

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C S Schou, H Mortensen. 1989-08-14. [Fractionated P-bilirubins].. https://pubmed.ncbi.nlm.nih.gov/2773134/

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Solvent partition of 14C-unconjugated bilirubin to remove labeled polar contaminants.

Polar derivatives contaminating unconjugated bilirubin (UCB) are customarily extracted with weak alkali. As UCB degrades rapidly at alkaline pH, regeneration of polar derivatives may limit purification. This problem is especially important when trying to eliminate radiolabeled contaminants from 14C-UCB. As polar derivatives of UCB should have a much greater aqueous to CHCl3 partition ratio (PR) than UCB even at neutral pH, where degradation of UCB is minimal, 14C-UCB in CHCl3 was serially extracted with an aqueous buffer at pH 7.0 to determine whether labeled derivatives could be preferentially removed. A single extraction of customarily purified 14C-UCB removed 0.18+/-0.06% of the radioactivity as labeled derivatives. Subsequent serial extractions yielded a stable, 67% lower 14C-PR with only 0.03% of radioactivity as labeled derivatives. Reverse-phase high-performance liquid chromatography (HPLC) of phases from later extractions revealed, however, that up to 1.1% of the disintegration per minute (dpm) in CHCl3 phases and up to 50% in aqueous phases were polar impurities. HPLC of partition phases spiked with purified 14C-UCB revealed that these impurities derived from incomplete extraction of the least polar impurities and their regeneration during HPLC. In the dark under argon, 14C-UCB in CHCl3or dimethyl sulfoxide (DMSO) solution degraded very slowly to polar derivatives. Extraction of impurities from a solution of 14C-UCB in CHCl3 is best done using pH 7.0 buffer, with removal of over 80% of the labeled contaminants remaining after customary purification by alkaline extraction.

Bilirubin↗