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Clinical Chemistry 7: 231-240, 1961;
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Clinical Chemistry, Vol 7, 231-240, Copyright © 1961 by the American Association for Clinical Chemistry

Studies on the Determination of Bile Pigments

III. Standardization of the Determination of Urobilinogen as Urobilinogen-Aldehyde

Richard J. Henry 1, S. L. Jacobs 1, and Sam Berkman 1

1 Bio-Science Research Foundation, 12330 Santa Monica Blvd., Los Angeles 25, Calif.

Several preparations of urobilin and stercobilin were investigated with regard to their value as standards for the determination of urobilinogen and urobilin in biologic specimens. Products of commercial origin were found to be impure and therefore unsuitable for this purpose. Absorptivities were measured for solutions of the pigments in 3% HCI in methanol and for the chromogens, obtained by ferrous hydroxide reduction, in the "quantitative" and "semiquantitative" methods for urobilinogen. Ascorbic acid was shown to be effective as a protective agent in filtrates from reduction mixtures.

Spectra of urobilinogen-aldehyde, phenolsulfonphthalein, Pontacyl dye mixture and phenolphthalein were compared. Phenolsulfonphthalein is recommended as the artificial standard of choice. The independence of this procedure from instrumental considerations, within limits, is demonstrated with the use of this dye.

Submitted on October 22, 1960






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Copyright © 1961 by the American Association for Clinical Chemistry.