Bilirubin Direct LS Jendrassik-Grof method

Bilirubin Direct LS Jendrassik-Grof method
80 – 85 % of bilirubin Direct LS originates on degradation of hemoglobin with the other 15 – 20 % being derived from cytochrome, myoglobin and catalases. Unconjugated bilirubin, which binds to plasma albumin, is produced in the course of degradation in the reticuloendothelial system, liver Kupffer cells, spleen and bone marrow. Unconjugated (primary, indirect, water-insoluble) bilirubin is soluble in lipids and toxic. With the aid of the glucuronyl transferase enzyme, bilirubin is conjugated primarily by glucuronic acid in the microsomes of hepatic parenchymal cells. In contrast to unconjugated bilirubin, conjugated (secondary, direct) bilirubin ibis soluble in water, and is excreted via the kidneys. Bilirubin assays are suitable for evaluating the degree of severity of icteric clinical symptoms as well as for monitoring and objectively assessing these symptoms. Distinguishing between direct and indirect bilirubin is a valuable aid in the differential diagnosis of different forms of jaundice. A direct bilirubin value of < 20 % total bilirubin is an indicator of jaundice of pre-hepatic origin. This value can increase to > 50 % in hepatic and post-hepatic jaundice.

Intended use
Quantitative determination of direct bilirubin in serum, heparinized plasma or EDTA plasma by the Jendrassik- Gróf method.

Test principle
Jendrassik-Gróf method
The azobilirubin produced by the reaction between bilirubins and the diazonium salt of sulfanilic acid shows maximum absorption at 555 nm in an acid medium. The intensity of the colour produced is proportional to the quantity of bilirubin which has reacted. In the absence of an accelerator, only conjugated bilirubins react.

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