4-hb. Estimation

Practical Physiology 4th Lab.

Estimation of Haemoglobin (Hb. Estimation) [email protected]

Lab. no. 4: Hb. Estimation Procedure:

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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. [email protected]

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). [email protected]

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.

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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 photocolourimeter.

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