List down the different techniques used in Hemoglobin determination; include the principle and the advantages and

disadvantages of each test.

Hb Determination Principle Advantages Disadvantages

Techniques
1. Cyanmethemoglobin • Blood is diluted in a
Method
2. Sahli’s Method • Blood is mixed with N/10
3. Automated Analyzer • Electrical
• All forms of Hb except • Diluted blood has to
solution containing Sulfhemoglobin are stand for a period of
potassium cyanide and converted to HiCN. time to ensure complete
potassium ferricyanide. • Cyanmethemoglobin conversion of Hb.
The latter converts Hb to solution is stable and it’s • Potassium cyanide is a
methemoglobin which is color does not fade with poisonous substance
converted to time so readings may not and that is why
cyanmethemoglobin be taken immediately. Drabkin’s solution must
(HiCN) by potassium • Absorbance may be never be pipetted by
cyanide. The absorbance measured soon after mouth.
of the solution is then dilution. • The rate of conversion of
measured in a • A reliable and stable blood containing
spectrophotometer at a reference standard is carboxyhemoglobin is
wavelength of 540nm or available from World slowed considerably.
in a colorimeter using a Health Organization for Prolonging the reaction
yellow green filter. direct comparison. time to 30min can
overcome this problem.
• Easy to perform • Less accurate.
HCl resulting in the • Quick • All Hbs are not
conversion of Hb to acid • Inexpensive converted to acid
hematin which is brown • Can be used as a bedside hematin and hence the
in color. The solution is procedure value of Hb obtained is
diluted till its color • Does not require less than the actual
matches with the brown technical expertise value.
colored glass of the • The color of acid
comparator box. The hematin develops
concentration of Hb is slowly.
read directly. • Individual variation in
matching of color is
seen.
impedance • Accurate and reliable • Laboratory Dependent
whole blood is passed • Expensive
between to electrodes
through an aperture so
narrow that only one cell
can pass through of a
line.

References:

Estimation of Hemoglobin. (n.d.). Hematology and Blood Bank Technique Module. Pages 19 – 22.

Lotspeich-Steininger, C. A., Stiene-Martin, E.A., Keopke, J.A. (1992). Clinical Hematology Principles, Procedures,
Correlations. J.B. Lippincott Company. Page 109.