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Presentation for Hb. estimation.Demonstration
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.
Haemoglobin
estimation method
Manual
Cyanmethaemoglobin
method
Acid Haematin
method
Automate
d
Coulter counter
Haemoglobin is converted to acid haematin
by N/10 HCl, the resulting brown colour
solution is compared with standard brown
glass reference blocks.
The intensity of the brown colour depends
on the amount of acid haematin produced,
which in turn depends on the amount of
haemoglobin in the blood sample.
Instruments in Sahli’s Haemoglobinometer
The main parts of which are
a graduated glass tube,
colour comparators box,
glass stirrer and Sahli’s
pipette to measure 20 µl
of blood.
The tubes commonly used
are with graduations in
percent on one side and
grams per 100 ml on the
other.
The colour comparators are
made of brown coloured
glass and some better
instruments have glass
prisms.
Haemoglobinometer box
Sahli’s pipette
Comparator
Box
Graduated
glass tube
Stirrer
Reagents:
• N/10 Hydrochloric acid
(HCl)
• Distilled water for dilution.
• Fresh blood or Blood
anticoagulated with EDTA
Procedure
 Place N/10 HCl in the tube up
to the lowest mark i.e. 2gm%
 Draw blood up to 20 mm mark
in the pipette and transfer it to
the acid in the tube.
 Rinse the pipette well by
drawing up the acid and re
expressing it. Mix the acid and
blood by shaking the tube well.
Allow it to stand for at least 10 minutes - to allow brown colour to develop due to
the formation of acid haematin.
 95% of Hb is converted at the end of 10 minutes, 98% of the colour develop at
the end of 20 minutes, and the maximum colour is reached after about 1 hour.
 Now dilute the solution with distilled water drop by drop with continuous
mixing, using the glass rod provided.
Purpose of estimating haemoglobin
 To detect the oxygen carrying capacity of
blood.
 The result assists in detecting diseases, which
causes a deficiency or excess of haemoglobin.
 Studying changes in haemoglobin
concentration before or after operations and
blood transfusions.
 To detect anaemia and its severity and to
monitor an anemic patients response to
treatment.
 To check haemoglobin level of blood prior to
donating blood.
 To calculate red cell indices.

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Presentation for Hb. estimation.Demonstration

  • 2. 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.
  • 4. Haemoglobin is converted to acid haematin by N/10 HCl, the resulting brown colour solution is compared with standard brown glass reference blocks. The intensity of the brown colour depends on the amount of acid haematin produced, which in turn depends on the amount of haemoglobin in the blood sample.
  • 5. Instruments in Sahli’s Haemoglobinometer The main parts of which are a graduated glass tube, colour comparators box, glass stirrer and Sahli’s pipette to measure 20 µl of blood. The tubes commonly used are with graduations in percent on one side and grams per 100 ml on the other. The colour comparators are made of brown coloured glass and some better instruments have glass prisms. Haemoglobinometer box Sahli’s pipette Comparator Box Graduated glass tube Stirrer
  • 6. Reagents: • N/10 Hydrochloric acid (HCl) • Distilled water for dilution. • Fresh blood or Blood anticoagulated with EDTA
  • 7. Procedure  Place N/10 HCl in the tube up to the lowest mark i.e. 2gm%  Draw blood up to 20 mm mark in the pipette and transfer it to the acid in the tube.  Rinse the pipette well by drawing up the acid and re expressing it. Mix the acid and blood by shaking the tube well.
  • 8. Allow it to stand for at least 10 minutes - to allow brown colour to develop due to the formation of acid haematin.  95% of Hb is converted at the end of 10 minutes, 98% of the colour develop at the end of 20 minutes, and the maximum colour is reached after about 1 hour.  Now dilute the solution with distilled water drop by drop with continuous mixing, using the glass rod provided.
  • 9. Purpose of estimating haemoglobin  To detect the oxygen carrying capacity of blood.  The result assists in detecting diseases, which causes a deficiency or excess of haemoglobin.  Studying changes in haemoglobin concentration before or after operations and blood transfusions.  To detect anaemia and its severity and to monitor an anemic patients response to treatment.  To check haemoglobin level of blood prior to donating blood.  To calculate red cell indices.