Drabkin’s method hemoglobin estimation procedure | Cyanmethemoglobin method PPT.

Drabkin’s method hemoglobin estimation procedure.Cyanmethemoglobin method PPT.

Today I am going to tell you about”

Drabkin’s method hemoglobin estimation procedure.Cyanmethemoglobin method PPT “hemoglobin test procedure and it’s calculation Cyanmethemoglobin method is also known as colorimetric method because this test is done by colorimeter. If you want to know about hemoglobin and hemoglobin all test method can you read here from this link.

What do you mean by Cyanmethemoglobin ?

In the Drabkin’s method or cyanmethemoglobin method or colorimetric method of hemoglobin estimation, hemoglobin is oxidised to methemoglobin by potassium ferricyanide ,which reacts with cyanide ions of potassium cyanide to form cyanmethemoglobin.This is the
internationally recommended method for determining hemoglobin .

Drabkin’s method hemoglobin estimation procedure.Cyanmethemoglobin method PPT

Principle of cyanmethemoglobin method.

Principle – Blood is diluted in a solution containing potassium cyanide and potassium ferricyanide. The latter converts HB to methemoglobin which is converted to cyanmethemoglobin by potassium cyanide. The observance of the solution is then measured in the a spectrophotometer at a wavelength of 540 nm(nanometer) or in a calorimeter using a yellow green filter.

Required equipment-

HB pipette
spectrophotometer
Drabkin’s reagent

The composition of Drabkins solution –

In Drabkin’s solution

PH-7.0-7.4 which contains

Potassium cyanide -50mg
Potassium ferricynide -200mg
Potassium dihydrogen phosphate -140mg
Non ionic detergent -1ml
Distilled water -1L

Test procedure of cyanmethemoglobin –

First we take three test tube which is labelled as b-blank, S-standard and T- test. and then we take 5 ml Drabkin’s solution in all three test tube. After that we take 20 microliter (0.02ml) distilled water in “B-Blank ” tube,20 microliter standard in “S-standard”tube and 20 microliter blood (EDTA blood) in ” T-test” tube. If you have any difficulty in this please see this

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Reagent	Blank	standard	test
Drabkin’s solution	5 ml .	5 ml.	5 ml.

Distilled water	20 microliter	———	————-
Standard	———	20 microliter	————

Blood.	——-	——-	20 microlitre.

After that mix all the tube and take OD (optical density) against distilled water at 540 nm (nanometer) Yellow or green filter.

After that not the all result and calculate this.

Calculation–

Calculation =OD of test /OD of standard X concentration of standard (15.06 gm/dl).

Normal range –

In Male – 14-18 gm% or gm/dl

Female – 12-16 % or gm/dl

Always remember this-

* don’t be pipette Drabkin’s solution by mouth.

the Dragon solution should be clear and pale yellow in colour when measured against water as a blank in a spectrometer at a wavelength of 540 NM the absorbent must be zero.
The solution is unstable if exposed to light and can be stored at room temperature in brown Boro silicate bottles for several months.
However if the room temperature is higher than 30 degree Celsius the solution should be stored in a refrigerator but about to room temperature before use the solution must never be frozen the pH of the solution must be checked every month.

Advantage of this method –

Visible error is not there as no colour matching is required.
Give accurate result
Cyanmethemoglobin solution is a stable and its colour doesn’t fade with time so readings may not be taken immediately.
Absorbance may be measured soon after dilution.
A reliable and stable reference standard is available from World Health Organisation for direct comparison.

Disadvantage of this method-

Diluted blood has to stand for a period of time to ensure complete conversation of Hb.
Potassium cyanide poisonous substance and that is why Drabkin’s solution must never be pipetted by mouth.
Abnormal plasma proteins cause turbidity when blood is diluted with Drabkin’s solution.
A high leukocyte count also causes turbidity on dilution of blood. centrifuging the diluted blood can help overcome the turbidity.

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