HEMOGLOBIN DETERMINATION

1. HEMOGLOBIN
DETERMINATION
Kalpesh Chhabhaiya
Enroll-100420117062

2. What is hemoglobin test ?
• A hemoglobin test measures the amount of
hemoglobin in your blood.
• Hemoglobin is a protein in your red blood cells
that carries oxygen to your body's organs and
tissues and transports carbon dioxide from your
organs and tissues back to your lungs.
• If a hemoglobin test reveals that your hemoglobin
level is lower than normal, it means you have a
low red blood cell count (anemia). Anemia can
have many different causes, including vitamin
deficiencies, bleeding and chronic diseases.

3. I. COLORIMETRIC METHOD
A. Direct visual colorimetric Method
• Tall quits method
• Dare’s Hemoglobin meter
• Acid Hematin method
• Alkaline Hematin method

4. B. Photoelectric colorimetric method
1. Oxyhemoglobin method
• Measures normal hemoglobin
• Used 0.007 N NH4OH or 0.1% Na2Co3
• Read with the wavelength at 540 nm

5. 2. Cyanmethemoglobin (HiCN)
• Also known as hemiglobin cyanide or
ferrihemoglobin cyanide
• All forms of hemoglobin are measured except
sulfohemoglobin
• Uses Drabkin’s solution
• Potassium ferricyanide
• Potassium cyanide
• Dihydrogen potassium phospate
• Distilled water
• PH = 7.0-7.4
( blood capacity )
• Used sahli pipet= (0.02ml or 20 micro liter)

6. PROCEDURE
• Place 5ml of Drabkin’s reagent into a testube
• Get 0.02 ml of whole blood using sahli pipet
• Place the 0.02 ml of blood in to drabkin's reagent
through rinsing it.
• Mix and let it stand for 10 minutes
• Read in a spectrophotometer at 540 nm.

7. II. Specific gravity method/Gravitational method
CUSO4 method
• Specific gravity of copper sulfate = 1.053 with an
hemoglobin equivalent of 12.5 gm%
• Mass blood
Procedure
• Collect blood sample
• Drop a blood into the solution
• Observe the activity of the blood
• Within 12 seconds, describe how the drop of
blood behaves.

8. III. Gasometric Method
• Indirect method
• Based on the assumption that 1gm Hb can
carry approximately 1.34 ml O2.
IV. Chemical Method
• Indirect method
• Based on the assumption that 1gm Hb contains
approximately 3.47 mg iron.

9. Electric Medical Equipment MHX-2
Full Automated Hemoglobin test

10. Interpretation :• Maintain = equal to 1.053 = 12.5 gm%
• Sink = > 1.053 = >12.5 gm%
• Float = < 1.053 = <12.5 gm%
Technical error
• Pipetting
• Shaking the pipet
• Charging the counting chamber
• Application of cover slip
• Counting of the cells
• Computation
• Reporting of results.

11. Never leave that till tomorrow which you can do
today.
Thank you.
Kalpesh Chhabhaiya
Enroll-100420117062