Method and stable diazo reagent for detecting bilirubin

Chemistry: analytical and immunological testing – Heterocyclic carbon compound – Hetero-n

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436903, 534559, G01N 3372

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active

046720413

ABSTRACT:
An improved reagent combination of the type comprising a first solution comprising an aryl amine and an acid; and a second solution comprising a nitrite compound. The regent combination is characterized in that is further comprises a salt, wherein the salt is present in any preselected distribution in the solutions.
Also, an improved method for colorimetrically determining bilirubin in a sample comprising mixing an aliquot of the above first solution with an aliquot of the above second solution to form a stable diazo reagent; contacting a sample to be assayed with a total bilirubin accelerator buffer system to form a prereaction medium; contacting the prereaction medium with the stable diazo reagent; and measuring the color of the azobilirubin formed form the coupling reaction of the stable diazo reagent with the bilirubin present in said sample.
In addition, an improved method for producing a diazo reagent of the type comprising reacting an aryl amine with a nitrite compound in the presence of an acid. The method is characterized in that the reaction also takes place in the presence of a salt.

REFERENCES:
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patent: 2827449 (1958-03-01), Kesler
patent: 3511607 (1970-05-01), Green
patent: 4030885 (1977-06-01), Das
patent: 4038031 (1977-07-01), Lam
patent: 4078892 (1978-03-01), Steinbrink, Jr.

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