A Quantitative Fractionation Method for Bilirubin in Human Bile

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Abstract
The main ingredient of the pigmented calcium stone is calcium bilirubinate. This substance has been known to be formed by hydrolysis of bilirubin glucuronide, the commonest type of bile pigment in bile, by [beta]-glucuronidase of bacterial origin and subsequent combination of the resulting free bilirubin with calcium. However, it also seems to be possible that a portion of bilirubin exists in bile as labile molecular compounds or conjugates which, in some instances, are able to combine with calcium to be precipitated as calcium bilirubinate. To evaluate the latter substances quantitatively, the authors established a systematic fractionation of bile pigments in bile using several solvents, and isolated some fractions consisting of labile bilirubin conjugates with phospholipids, proteins and bile acids.