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Practical Physiology

4th Lab.

Estimation of
(Hb. Estimation)
Lab. no. 4: Hb. Estimation
• In the graduated tube of the Sahliś
haemoglobinometer add (N/10 HCl) up to (2 g%)
mark. Clean the ball of the left ring finger with spirit
& let it dry.
• Give a moderately deep prick with a sterilized
lancet, let a large drop of blood collect over the
finger and then suck in the pipette of the
haemometer up to 20 mm3 (= 20 μl ) mark.
• Wipe the tip of the pipette and immediately transfer
the blood into the graduated tube. Mix the blood and
the HCl & let it remain for (5-10) min.
Lab. no. 4: Hb. Estimation
3.Brownish coloured acid haematin is formed by the
action of HCl on Hb.
4.Dilute it with D.W mixing it with a stirrer till the colour
matches with the standard colour (coloured plates)
of the haemoglobinometer.
5.Near the end point take three readings: when the
colour of the mixture is slightly darker, when the
colour is exactly similar and when the colour is just
slightly fainter.
The mean of the last 3 readings is taken as the true
reading of haemoglobin concentration.
-Note: the reading is taken on ( g % ) scale(=g/dl).
Lab. no. 4: Hb. Estimation
Normal ranges:
Just after birth (16-22) g %.
adult male (14-18) g %.
adult female (12-15) g %.

Physiological causes of increased Hb level :

1. Infancy.
2. High altitude.
Lab. no. 4: Hb. Estimation
Photometric method:

It is also used to determine Hb concentration. It is the

most reliable method since subjective errors due to
visual comparison are eliminated. Certain chemicals
can react with Hb forming colour compounds and
the intensity of colour is read in a photo-