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Hb Blood Test Method / hgb blood test: Complete hemoglobin estimation procedure with results. we have used Sahil's method to estimate blood Haemoglobin level i.e. Normal Hb level. Haemoglobin Count in Blood is also called Hb count or #haemoglobintest.
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Principle of Haemoglobin Test Procedure:
Blood is mixed with N/10 HCl resulting in the conversion of Haemoglobin to acid hematin which is brown in color. The solution is diluted till its color matches with the brown-colored glass of the comparator box. The concentration of Haemoglobin is read directly.
Haemoglobin Estimation with Sahli’s/acid hematin Method:
Equipment required for Hb Count:
Hemocytometer: amzn.to/3cQIx9q which consists of a comparator box that has brown colored glass on either side Hb pipette which is marked up to 20mm3(0.02ml blood) Tube with markings of Hb on one side
glass rod dropper.
Sterile Cotton: amzn.to/3gC0rhm
Reagent Bottles: amzn.to/3xvRBZj
Reagents required for Hb test:
N/10 HCl
Distilled water
Sample: Venous blood collected in EDTA as described earlier
The procedure of Hb Count:
1. Add N/10 HCl into the tube up to mark 2g%
2. Mix the EDTA sample by gentle inversion and fill the pipette with 0.02ml blood. Wipe the external surface of the pipette to remove any excess blood.
3. Add the blood into the tube containing HCl. Wash out the contents of the pipette by drawing in and blowing out the acid two to three times. Mix the blood with the acid thoroughly.
4. Allow to stand undisturbed for 10min.
5. Place the haemoglobinometer tube in the comparator and add distilled water to the solution drop by drop stirring with the glass rod till its color matches that of the comparator glass. While matching the color, the glass rod must be removed from the solution and held vertically in the tube.
6. Remove the stirrer and take the reading directly by noting the height of the diluted acid hematin and expressing in g%.
Advantages: blood hemoglobin method
Easy to perform
Quick
Inexpensive
Can be used as a bedside procedure
Does not require technical expertise
Disadvantages: Hemoglobin Test Procedure
Less accurate.
All haemoglobins (oxyhemoglobin, sulphemoglobin) are not converted to acid hematin and hence the value of Hb obtained is less than the actual value.
The colour of the acid hematin develops slowly.
Colour of acid hematin fades with time and dilution must be done exactly after 10 min when the colour development is maximum
Individual variation in the matching of colour is seen.
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