Clinical Chemistry
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Clinical Chemistry 20: 447-453, 1974;
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Clinical Chemistry, Vol 20, 447-453, Copyright © 1974 by the American Association for Clinical Chemistry

Detection and Measurement of Total Bilirubin in Serum, with Use of Surfactants as Solubilizing Agents

Frederick C. Pearlman 1 and Robert T. Y. Lee 1

1 Diagnostics Reagents and Equipment Department, Science Products Division, Mallinckrodt Chemical Works, 2703 Wagner Place, Maryland Heights, Mo. 63042.

We describe a new method for measuring total bilirubin in serum. Nonionic, cationic, or anionic surfactants can be used as solubilizing agents to promote the diazo coupling of indirect-reacting bilirubin. A representative surfactant—azobilirubin system is illustrated, in which absorbance is maximum at 560 nm and linear to a concentration of 200 mg of bilirubin per liter. At pH 2.5 and in the presence of Du ponol (an anionic surfactant), bilirubin is completely coupled within 6 mm at 37 °C. All ingredients required for an assay are combined in two dry reagents, which are stable for a year at room temperature. Sensitivity is comparable to that of procedures routinely used in clinical laboratories; precision, linearity, and stability of the reagents in solution are excellent. Results for fresh sera correlated well with those obtained by the Jendrassik—Grof method. Hemolysis is minimized and turbidity eliminated.


Key Words: Jendrassik—Grof method compared • error evaluation as compared with four methods

Submitted on August 28, 1973
Accepted on January 21, 1974




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Copyright © 1974 by the American Association for Clinical Chemistry.