Blood analysis Lab # 4

Hemoglobin (Hb) Characteristics: 1-Hb is a associated protein - Non-protein part → Heme (iron protoporphyrin) - Protein part → Globin 2-Hb is a chromoprotein i.e it gives the blood its red color Function: O2 transport Hb + O2 → Oxyhemoglobin

Determination of Hemoglobin (Hb) (Cyanmethemoglobin Technique) Principle Hb + Ferricyanide → met-Hb Met-Hb + cyanide → cyanomethemoglobin Procedure Drabkin reagent + 0.02 blood read abs. at 546 5 min.

Cont. Calculation Normal levels: Hb conc. (gm/dl) = 36.77 x A M: 14-18 g/dl F: 12-16 g/dl

Interpretation of the results ↓Hb → Anemia Causes of anemia: 1- ↓ Production of Hb a- Iron deficiency b- Vitamins deficiency e.g. vit B12, vit B6 & folic acid c- Protein deficiency

Interpretation of the results 2- ↑ Hb destruction a- Hemolytic anemia b- Heavy metal poisoning e.g. lead c- Infections e.g. malaria ↑ Hb → Polycythemia Causes: - High altitudes - CO intoxication - Pulmonary diseases e.g. emphysema

Bilirubin Bilirubin is the end product of Hb destruction

Bilirubin bilirubin + glucouronic acid →water soluble bilirubin Types of bilirubin 1- Conjugated = soluble =direct bilirubin + glucouronic acid →water soluble bilirubin 2- Unconjugated = insoluble = indirect

Bilirubin: cont.

Bilirubin: cont. Total bilirubin= direct + indirect Normal values: Total bilirubin: 0-1 mg/dl Direct bilirubin: up to 0.25 mg/dl Indirect bilirubin: up to 0.75 mg/dl

Clinical significance Hyperbilirubinemia indicates Jaundice Causes of jaundice 1- Pre-hepatic (↑ indirect bilirubin) a- Hemolytic anemia b- Incompatible blood transfusion 2- Hepatic (↑ indirect bilirubin) a- ↓ glucouronyl transferase enz. b- Viral hepatitis & liver cirrhosis 3- Post hepatic (↑ direct bilirubin) common bile duct obstruction

Iron Iron physiology Iron forms: Ferrous → in Hb & oxyHb Ferric → in cyanmetHb Iron physiology

Iron containing compounds 1- Oxygen carrying chromoproteins: e.g. Hb & myoglobin 2- Enzymes: e.g. cytochromes, catalase, peroxidase

Iron Absorption Storage Duodenum & jejunum Iron is stored in ferric form

Determination of iron Principle 1- Acidification; transferrin-Ferrric HCL transferrin + ferric ion 2- Reduction: ferric ion + thioglycolic acid ferrous ion 3- Coloring: ferrous ion + bathophenanthrolene pink color

Iron ↑ iron ↓ iron 1- ↑ RBCs destruction 2- ↑ iron absorption Normal value 60 – 160 mg/dl Clinical significance ↑ iron ↓ iron 1- ↑ RBCs destruction 2- ↑ iron absorption 3- Iron overload 4- Liver diseases 1- Decreased dietary iron intake 2- Blood loss 3- Increased demand e.g. pregnancy & children