1. By Dr. Manal Basyouni Dr. Manal Basyouni

2. Heme consists of a Porphyrin ring coordinated with iron. It is found mainly in hemoglobin but also present in myoglobin and cytochrome. Normal Hb Values: Male: 14 -18 g%. Female: 12 -16 g%. Sample: whole blood (anticoagulant as EDTA may be added). Dr. Manal Basyouni

3. : Principle 1 - Hb is oxidized HB by Potassium ferricyanide {K Fe (CN)} into Met-haemoglobin HB (Fe +++ ). HB (Fe ++ ) + K Fe +++ (CN) HB (Fe +++ ) + K Fe +++ (CN ) 2 - Met-haemoglobin is converted into Cyanmet- haemoglobin by Potassium cyanide. HB (Fe +++ ) + K CN HB (Fe +++ ) CN 3- It m easured by photometer at 540 nm. 4- HB conc. = A sample 36.77 (g%) Or use standard curve for Cyanmet-haemoglobin Dr. Manal Basyouni

4. : Principle of assay (Colorimetric Method) HB (Fe ++ ) (Fe +++ CN) Dr. Manal Basyouni Potassium ferricyanide Met-haemoglobin HB (Fe +++ ) Cyanmet-haemoglobin HB (Fe +++ ) CN Measured by photometer at 540 nm Potassium cyanide (K CN)

5. Diagnostic Significance of Hb HB conc. is decrease in : 1. Hemolytic anemias, e.g. thalassemias, Sickle cell anemia. 2. Iron deficiency anemia. 3. Megaloblastic anemia (folic acid or vit. B12 deficeincy). 4. Hemorrhagic anemias e.g. Vit. K deficiency, menorhagia, ankolystoma infection). Dr. Manal Basyoun i

6. HB conc. is increase in : 1- Polycythemia (erythrocytosis). Primary cause: Polycythemia Vera. Secondary cause: cyanic heart diseases, smoking, tumors secreting erythropoitin. 2- Decrease blood volume: e.g. dehydration, vomiting, diarrhea, use of diuretics. Dr. Manal Basyouni