THEME Nr

THEME Nr.29

Experiment Nr.1 Hemoglobin assay in blood.

Methods principle: Hemoglobin is oxidised by potassium ferricyanide (K3Fe(CN)6) to methemoglobin, which will change the color of the solution from yellow to pink.

Procedure: Mix in a test-tube 5 ml of transforming solution (contains K3Fe(CN)6) and 0,02 ml of blood. Incubate 10 min and measure the extinction of the experimental solution in 10 mm cuvettes (green light) using transforming solution as a control.

Normal values: - 132-164 g/l;

115-145 g/l.

Clinical value: The content of hemoglobin decreases in anemias and increases in dehydration and erythremias.

Rezults:

Conclusion:

Self-preparing questions:

1. Functions of the blood.

2. General chemical composition of the blood.

3. Chemical composition of the red blood cells (erythrocytes): hemoglobin, non-heminic proteins, enzymes, minerals.

4. The metabolism of the nature erythrocytes:

-glycolysis and the 2,3-bisphosphoglycerate pathways,

-the pentose-phosphate pathway and its role in the antioxidative defence,

-ATP production.

5. Chemical composition and the metabolical peculiarities of the white blood cells.

6. Chemical composition and metabolical peculiarities of the platelets.

7. Hereditare disorders of the blood cells metabolism (deficiency of glucose-6-phosphate dehydrogenase; pyruvate kinase; chronic granulomatous disease; type I leukocyte adhesion deficiency; hemophilia A.

Self-testing questions:

1. Why NADPH is a key compound in the red blood cell metabolism?

2. What is the normal content of hemoglobin and erythrocytes? What conditions can change it? How?

3. What active species of oxygen are produced in the neutrophils? How? What is their role?

4. What clotling factors are produced by the platlets? What functions are they carring out?