Estimation of Haemoglobin in Shali’sMethod

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Estimation of Haemoglobin in Shali’sMethod

 

AIM: To determine the Hb content of the blood to calculate the O2 carrying capacity of blood to find out the iron content of the blood.

 

Principle: Matching the colour of the diluting blood in which Hb is converted in to acid haematin by treatment with N/10 Hcl a permanent standard coloured plates.

 

Apparatus: Sahil’s Haemometer. Haemoglobin pipette. N/10 Hcl. Distilled water stirrer Lancet, Cotton, Spirit.

 

Procedure: Take N/100 Hcl in graduated tube upto the mark 10 on the % a scale of the lowest mark. Sterlise the finger with cotton and spirit prick the finger by lancet and draw the blood in to the Haemoglobin pipitte up to the mark 20Cu, mm wipe away any any blood adhering the pipette. Transfer the blood to the graduated tube immediately repeated filling and emptying the pipette with Hcl in the graduated tube. Mix the blood with HCL by the stirrer and wait for five minutes the clear brown colour of the mixture is due to formation of acid haematin.

 

                Dilute the mixture with distilled water drop by drop stir. Care should be taken that no froth or air bubbles result. Proceed on diluting till the colour of the mixture exactly matches the colour of the standard plates. Take the reading of the graduated tubes as Gms of Haemoglobin / 100ml of blood.

 

Normal Value:

                In male 15 to 17 Gms/100 ml of blood

                In female 13 to 15 Gms/100 ml of blood

 

Variations: Increase in the Hb content occurs in the following conditions.

 

Physical:               1) More in new born infants about 20 to 23 G/100 ml

                                2) More in the afternoon 3) Increases during and after muscular exercise.

                                4) Increases in emotional state 5) Increases at hige altitude.

 

                In the above conditions the variation is due to alteration in the blood cell count.

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