HEMOGLOBIN

DETERMINATION
By: Tom Anthony A. Tonguia, RMT
Hemoglobin determination
◦ Determination of hemoglobin can be done separately or as a part of routine CBC.
◦ This is a standard and part of the automated instrumentation that includes cell
counts and a calculated hematocrit.
◦ Hemoglobin determination is relatively simple and can be performed quickly by the
laboratory.
Hemoglobin Measurement in the
laboratory
◦ Various methods of determining hemoglobin have been used through the years.
◦ Methods currently in common use include the specific gravity (gravimetric method)
technique, the cyanmethemoglobin method, and automated methods.
Gravimetric/Specific Gravity
Technique
◦ This is a qualitative measurement of hemoglobin.
◦ Is not routinely performed except as a method of screening blood donors.
◦ It gives only an estimate of hemoglobin concentration and requires no special
instrumentation.
◦ A drop of blood is allowed to fall into a copper sulfate (CuSO4) solution of a
particular density (specific gravity).
◦ If the drop falls through the solution rapidly, the specific gravity of the blood is
greater than the specific gravity of the copper sulfate while blood with a low
hemoglobin concentration does not fall rapidly.
Cyanmethemoglobin or
Hemiglobincyanide method
◦ Is the most widely used method of measuring blood hemoglobin.
◦ Blood is reacted with Drabkin’s reagent, which contains iron, potassium, cyanide
and sodium bicarbonate.
◦ The Drabkin’s and the hemoglobin combine to form a very stable colored end
product, cyanmethemoglobin, also called hemoglobincyanide (HiCN).
◦ Because most hemoglobin reagents contain hazardous chemicals such as cyanide
or azide, care must be taken when performing the test.
Automated method
◦ Most instruments used for other hematology analyses, such as cell counters, also
perform hemoglobin determinations.
◦ Most automated counters measure hemoglobin by a modification of the manual
HiCN method with cyanide reagent or with a non hazardous chemical such as
sodium lauryl sulphate, which avoids possible environmental hazards from disposal
of large volume of cyanide containing waste.
◦ Modifications include alterations in the concentration of reagents and in the
temperatu