Use of the caffeine reagent in direct spectrophotometry of bilirubin.

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Abstract
The molar absorptivity of unconjugated bilirubin in caffeine reagent is independent of the protein matrix. This finding, together with the simplicity of a dilution step for direct spectrophotometry, will improve the calibration methods of bilirubin and make them more nearly accurate. This is encouraging for the development of a new method for bilirubin determination in neonates; moreover, the caffeine reagent has a clearing influence on the turbidity of human sera. These findings should also be important for standardization, especially because the method of Jendrassik and Gróf is also protein-independent. Therefore, the introduction of one reliable, inexpensive, "universal" standard of bilirubin in bovine serum albumin will be of importance for both methods.