The red-violet pigment formed by the condensation of diazotized sulfanilic acid with bilirubin in the van den Bergh reaction.
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In the latter, the absorbance of Hb at low bilirubin concentrations overcompensates for decreased absorbance caused by the destruction of the azobilirubin, whereas at high bilirubin concentrations Hb does not compensate for this decrease.
The color (pink) intensity of the azobilirubin produced is proportional to the total bilirubin concentration.
Total bilirubin was analyzed by measuring azobilirubin after the addition of accelerators and diazotized sulfanilic acid (17).