Presentation for Hb. estimation.Demonstration

Hb estimation

1. DR. Kamla Choudhary HEMOGLOBIN ESTIMATION BY SAHLI'S METHOD & ACID HEMATIN METHOD (SAHLI’S HAEMOGLOBINOMETER )

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.

3. 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. Principle

4. 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 Dropper

5. Reagents: N/10 Hydrochloric acid (HCl) Distilled water for dilution. Fresh blood or Blood anticoagulated with EDTA N/10

6. 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.

7. 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.

8. 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.