Drabkin’s Solution for Hemoglobin, Preparation of Drabkin’s Solution
Table of Contents
Drabkin’s Solution for hemoglobin estimation
- There are commercially available kits, which is easy to make the solution.
- When blood runs in the hematology analyzer, these instruments also estimate hemoglobin.
Sample for hemoglobin estimation
- Whole blood is needed for hemoglobin estimation.
Manual Method for preparation of Drabkin’s solution
- Hemoglobin (Drabkin’s) solution can be prepared in the laboratory.
- Drabkin’s solution reagents needed are:
- Potassium ferricyanide = 200 mg
- Potassium cyanide = 50 mg
- Potassium dihydrogen phosphate = 140 mg
- Non-ionic detergent = 1 ml
- Distal water = Make up to 1000 ml (1 L)
Precautions to keep the Drabkin’s solution
- Keep the solution in a dark-colored bottle and in the dark to protect it from the light.
- If the solution is cloudy after adding the blood, in that case, centrifuge before the reading, this may be due to nonhemolyzed RBCs or globulins.
- Spectrophotometer cells should be fingerprint-free; otherwise, the reading will be high.
- Drabkin’s solution is a pale yellow clear fluid; it should not be used when it is cloudy.
The principle of Drabkin’s solution:
- Whole blood is diluted 1 into 200 dilutions with Drabkin’s solution, containing potassium ferricyanide and potassium cyanide.
- RBCs are hemolyzed by Drabkin’s solution.
- Now hemoglobin is oxidized, and its derivatives, except sulfhemoglobin, form methemoglobin in the presence of alkaline K- ferricyanide.
- The methemoglobin reacts with K-cyanide to form a very stable compound, cyanmethemoglobin, and this complex has maximum absorption at 540 nm.
Drabkin’s solution principle
How to estimate hemoglobin by Drabkin’s solution (Procedure):
- Take 20 microliter of blood + Drabkin 4 mL = 1 : 200 dilution.
- OR take 20 microliter of blood + Drabkin 5 mL = 1 : 250 dilution.
- Now mix well.
- Read within 6 hours of mixing the blood with Drabkin’s solution.
- Read on a spectrophotometer at filter 540.
- Read against the blank of Drabkin’s solution (Drabkin solution can be used as blank).
- Also, read the standard solution (12 G/dL) with the same dilution as the test sample.
- Read by the spectrophotometer; the reading is called optical density (OD)
Drabkin’s solution procedure for hemoglobin estimation
Hemoglobin calculation formula (OD = optical density)
Normal hemoglobin:
- Adult male = 14 to 18 g/dL
- Adult female = 12 to 16 g/dL
- 10 years old child = 12 to 14.5 g/dL
- 3 months old infants = 9 to 14 g/dL
- Newborn = 17 to 23 g/dL
Physiological variation of Hb:
- Strenuous physical exercise.
- There is a diurnal variation, with the highest level in the morning and low in the evening.
- High altitude increases the Hb concentration.
False causes of raised Hb:
- Hemoconcentration due to dehydration and burns.
- Immediately after hemorrhage.
- If taken during the I/V infusion, if it contains iron.
Questions and answers:
Question 1: What is the end result of KCN reaction for hemoglobin?.
Question 2: What is the physiological variation in the hemoglobin?
Go Back to Hematology
You shows the principle of drabkin solution but not show how we are apply this solution
Dear
As I have understood from your question, that you want the procedure by Drabkin’s solution. It is already given there. Or please explain your question.
I got good news
ok Thanks.
Can we take 2ml reagent and multiple the reading with 20
You have to keep the same dilution, that is very important.
Sir how to measure bilirubin ??
Dear nowadays nobody make reagents themselves. Ready-made kits are available in the market. Definitely you need a lab facility.
what is the function of potassium dihydrogen phosphate in drabkin’s solution ?
This has the buffering action.
Can drabkins reaction work with freeze dried hemoglobin instead?
I think if you thaw the frozen Hb, then try with control. That may work.
What’s the quality control of drabkin solution?
You can run known control of hemoglobin. These known controls are available from various companies.
what is the function of the non-ionic detergent in the drabkins solution ?
Please see the following link for your question.
https://patents.google.com/patent/US4341527A/en
May i ask sir, i read that a source of error when using the Drabkin’s reagent was a possible increase of globulins which can then be fixed using the dihydrogen potassium phosphate – is this true? and if so, how come what’s the principle behind it? Thank you !!
I have given diagrammatically the principle of Drabkin’s solution.
https://labpedia.net/haemoglobin-part-3-hemoglobin-solution-drabkins-solution-preparation/
Can we prepare the standard solution of cyanmethemoglobin process in lab…if it is not available commercially?
You can prepare Drabkin’s solution. I have described the procedure.
sir, what is OD in formula?
OD is the reading by the spectrophotometer, called optical density. Read the article again.
https://labpedia.net/drabkins-solution-for-hemoglobin-estimation/
Sir is Hb levels of 17.4 normal according to Drabkins method sir?
This is a higher limit. Please keep on checking Hemoglobin to rule out the possibility of polycythemia. It would be better to consult a hematologist.
Sir
Can hemoglobin react with potassium thiocyanate? Please
You can use sodium cyanide instead of potassium cyanide.
The procedure for commercially available cyanmethemoglobin standard soln is differ from the above mentioned one…
because if we use commercially available std. soln we should use directly if the soln. is in RT.
You can use a ready-made solution.