Manual micromethods for bilirubin determination in sera of adults and children and investigation of reasons for observed differences.

This study compared three micromanual methods for determining bilirubin concentration. The two microchemical methods for total bilirubin, a Jendrassik-Grof procedure and a Unopette® procedure, using dimethyl sulfoxide as an accelerator and protein solubilizer, gave comparable results in sera of adults and children. A microspectrophotometric method and the microchemical methods for total bilirubin gave similar results in plasmas of newborns with physiologic hyperbilirubinemia and in sera of older children with no hepatic abnormality. However, the microspectrophotometric method gave higher values in normal and hyperbilirubinemie adult sera. The results obtained with the Jendrassik-Grof and Unopette microchemical methods for direct bilirubin in sera of adults and children showed the values determined by the Unopette to be higher. Using the presently accepted normal range, this difference is significant enough to preclude recommendation of the use of the Unopette method for distinguishing normal from elevated levels of direct bilirubin. Direct bilirubin in newborn serum measured by the Unopette method is considerably higher than that measured by the Jendrassik-Grof method. An investigation to determine the reason for the difference in the direct bilirubin results indicated that the Unopette direct method measures diconjugated bilirubin in amounts similar to those measured by the Jendrassik-Grof method but significantly more monoconjugated bilirubin than the Jendrassik-Grof method.