Sahli's method of IHemoglobin and its measurement is known as acid haematin method.
To estimate the haemoglobin content in a sample of your own blood.
Haemoglobin present in a sample of blood is converted into acid hematin by addition of N/l 0HCl to the blood and its haemoglobin content is determined by matching the solution against non fading glass having a standard colour.
APPARATUS:Sahli's haemoglobinometer, distilled water, rectified spirit, cotton, and lancet.
1.The box has a space for keeping the tube in the middle.
The standard is non fading brown tinted glass, the colour of which is acid haematin.
The blood must be treated with 0.1 NHCL and diluting it 100 times.
2.The special diluting tube has a percentage scale on one side and agm/dl on the other side.
3.The capillary pipette has a 20mm3 marking.
The lowest mark in the diluting tube is where NI 10 HCI should be taken.The box has space for the diluting tube.The fingertip should be sterile using rectified spirit.To get a large drop of blood, make a quick prick.Don't have an air bubble when sucking the blood in the haemoglobin pipette.To remove blood from the tip and sides of the pipette, use cotton.The acid is taken in the tube.If you want to transfer the acid into the tube, rinse the pipette two or three times.So that haemoglobin gets converted to acid haematin, mix and keep it undisturbed for 10 minutes.After 10 minutes, add distilled water drop by drop and mixing the contents after each drop with the stirrer to match the colour of the standard.You can take the reading in gram scale and percentage scale.
1.There should be no air bubble or blood clot in the pipette.
2.Graduations on the tube should not affect colour matching.
3.Before colour matching and reading, the glass rod should be lifted.
4.Excess blood can be seen on the sides and tip of the pipette.
5.To note the time, transfer the contents immediately into the diluting tube.
6.If you take the reading without delay, the colour will deepen.
There are 1 individual variations in colour.
2.The standard fades in color with time.
3.Acid haematin is not formed by incomplete conversion of haemoglobin to other haemoglobins.
4.The colour matching will be affected by the presence of proteins and lipids in the blood.Other methods can be used.
The Alkali haematin method is a better method for estimating haemoglobin.It's even methaemoglobin.They are converted into alkali haematin.
The amount of gas bound by haemoglobin is what determines this.
1.Oxygen carrying capacity method.
It is possible to avoid visual comparison or colour matching with the naked eye by using photo electric colorimetry.Here haemoglobin may be estimated by the use of acid haematin and N10 HCI.
4.The method depends on the iron content of the blood.
1.Making use of gravity.
The haemoglobin content increases at high altitudes.Hypoxia increases haemoglobin content and stimulates erythropoiesis.The haemoglobin content increases after exercise.
Pathological variations include increased polycythemia and decreased anaemias.
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