HAEMOGLOBIN (HB) ESTIMATION


Method Used: Haemiglobincyanide (HiCN) Method.
Principle: whole  blood is diluted  1 in 201  dilutions  in a  modified   Drabkin’s  solution which contains potassium cyanide. The  red  cells are haemolyzed and the Hb  is  oxidized by the ferricyanide to methaemoglobin. This is converted by the cyanide to stable hemoglobin  cyanide (HiCN). Absorbance of  the  HiCN solution is read in  a  spectrophotometer at wavelength 540nm or in a filter Colorimeter using a yellow-green filter  (Cheesbrough, 2000) .


Procedure:
1.            measured  20µ1 (0.02ml)  of capillary blood on well –mixed venous blood  and  dispense it into  4ml  Drabkin’s  neutal diluting fluid
2.            stoppered the tube,  mixed  and left the diluted blood at room temperature  protected  from sunlight for  5-5minutes
3.            placed the yellow –green filter in a colorimeter or set at a wavelength at  540nm
4.            zero  the colorimeter with Drabkins fluid and read the absorbance of the  patients sample (CHEESBROUGHT, 2000)
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