Hemoglobin Estimation by Acid Hematin method

 

Estimation of Hemoglobin by acid hematin method

The hydrochloric acid (HCL) cause the hemolysis of Red blood cells and releases the hemoglobin and released hemoglobin is converted into acid hematin which is brown in color and matched with color standard.

 HCL + Hemoglobin (red) → Acid hematin (brown)

 

Requirements

• Ø  Hemometer
• Ø  Comparator
• Ø  Diluting tube
• Ø  Sahli pipette
• Ø  Dropper
• Ø  Cotton
• Ø  Stirrer
• Ø  1%  hcl solution
• Ø  Blood sample
• Ø  Distilled water

 

Procedure

• Ø  Take 1% HCL solution up to 2gm% mark perfectly into the clean and dry graduated diluting tube.
• Ø  Draw the well mixed anticoagulant blood in sahli pipette up to 20 mm mark.
• Ø  Wipe the side of the pipette by cotton to remove excessive blood.
• Ø  Then immediately transfer the blood into the graduated diluting tube and mix it well by gently shaking and keep it in comparator and leave it for 5 minute.
• Ø  After 5 minute add distilled water drop by drop and mix with stirrer.
• Ø  The mixing procedure is continued until the color of the comparator is matched with tube.
• Ø  After the matching the color of comparator removes it and takes result from tube.
• Ø  The result is express in gm%

SPECIES	RANGE(gm/dl)	AVERAGE(gm/dl)
Dog	12-18	15
Cat	8-15	12
Horse	8-15	12
Cattle	8-15	11
Sheep	9-12	11.5
Goat	5-15	10
Pig	10-16	13
Adult male	14-18
Adult female	12-16
Children	10-14