Separation and quantitative estimation of four bilirubin fractions from serum and of three bilirubin fractions from bile
Kuenzle, C.C.; Sommerhalder, M.; Rüttner, J.R.; Maier, C.
Journal of Laboratory and Clinical Medicine 67(2): 282-293
An improved method for the chromatographic separation and subsequent determination of bile pigments is given in detail. Reversed-phase partition chromatography of non-deproteinized serum and bile samples on silicone-treated kieselguhr columns yields four and three bilirubin fractions, respectively. Bilirubin fractions are eluted from columns with an alkaline aqueous solvent containing antioxidants. Bilirubin is estimated spectrophotometrically by its yellow color. The method satisfies the definite need for a quantitative determination of bilirubin fractions from serum and bile.