SAHLI'S ACID HAEMATIN METHOD :
Principle:
Haemoglobin is converted into acid haematin by the action of HCl. The acid haematin solution is furthur diluted with distilled water until its coolor matches with exactly that of permanent standard of comparator block. The Hb concentration is then read directly from the calibration tube.
Requirements: HCl solution, Sahli's Haemoglobinometer, pipette, distilled water.
Procedure:
Fill the Hb calibrated tube upto the mark 20 with 0.1N HCl bt means of a dropper. Draw blood upto 20μL mark in the Sahli's pipette. Adjust the blood coloumn carefully without bubbles. Wipe excess of blood on the sides of the pipette by using a dry piece of cotton. Blow the blood into the acid in the graduated tube, rinse the pipette well. Mix the reaction mixtures and allow the mixture to stand at room temperature for 10 minutes. Dilute the solution by adding distilled water drop by drop carefully and by mixing the reaction mixture until the color matches the color in the comparator. The lower meniscus of the fluid is noted and reading is noted in gm/dl.
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