A directly reacting fraction of bilirubin that is probably covalently bound to albumin (albumin-bound bilirubin) has recently been described. To determine its clinical importance we used a new high-performance liquid-chromatography technique to measure it in the serum of 200 patients with hyperbilirubinemia from various causes. Albumin-bound bilirubin was an important fraction (8 to 90 per cent) of total bilirubin in patients with hepatocellular and cholestatic jaundice as well as in patients with the Dubin-Johnson syndrome. It was not detected in normal volunteers, neonates with physiologic jaundice, or patients with Gilbert's disease or hemolysis. Thus, albumin-bound bilirubin appears in serum when hepatic excretion of conjugated bilirubin is impaired. It becomes a larger component of serum bilirubin as jaundice subsides, delaying resolution of this disorder and causing bilirubin to persist in plasma after it has disappeared from the urine.

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The angular correlation of the 173-247 keV gamma-ray cascade after the electron-capture decay of 11"In is strongly perturbed when the I-p-nitrophenylethylenediaminetetraacetate chelate of lllIn3+ is added to a solution containing rabbit antibody to dinitrophenyl groups. The radioactive chelate can be displaced by the addition of dinitrophenyllysine or unlabeled chelate. The average association constant between the antibody and the labeled chelate has been estimated from perturbed angular correlation measurements; this value is compared to the results of equilibrium dialysis. These experiments provide good evidence that information concerning macromolecular behavior can be obtained from perturbed angular correlation experiments that use chemically specific labels.