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Determination of Significance：
Red blood cells rupture and free hemoglobin (FHb) content increases in the plasma when hemolysis occurs in blood vessels. Plasma FHb concentration increases mildly to moderately in patients with autoimmune hemolytic anemia, sickle cell disease or thalassemia. The patients have hematuria symptoms who suffer from paroxysmal nocturnal hemoglobinuria, paroxysmal cold hemoglobinuria or unstable hemoglobin disease. It is important to detect FHb content for diagnosis of hemolytic diseases.
Measurement Principle：
Heme in hemoglobin has the catalytic function of peroxidase analogues. It could catalyze the oxidation of 4-aminoantipyrine and phenyl amines by H2O2 to form purple quinones. It has a characteristic absorption peak at 546 nm, and its color depth is directly proportional to FHb content.
Self Provided：
Spectrophotometer/Microplate reader, constant temperature foster box/water-bath, desk centrifuge, balance, transferpettor, micro glass cuvette/96 well plate, ice and distilled water.
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