This document describes various methods for determining hemoglobin levels through laboratory testing. It outlines direct visual and photoelectric colorimetric methods, including the oxyhemoglobin and cyanmethemoglobin (HiCN) techniques. Procedures are provided for the cyanmethemoglobin method using Drabkin's solution in a spectrophotometer. Additional methods covered include specific gravity/gravitational testing with copper sulfate and gasometric or chemical methods relying on hemoglobin's oxygen carrying or iron content properties. Interpretation of specific gravity tests and potential technical errors are also discussed.

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