Question
Asked 5th Dec, 2014

How to prepare EDTA as anticoagulant for human blood collection?

I have to collect human blood samples, but there are no commercial EDTA blood collection tubes available in my lab.
I would like to know how to prepare EDTA to use as an anticoagulant for blood collection, and I have 0.5M EDTA right now. Can I use this solution for preparing it or not? To which percentage should I prepare it?
Thank you so much.

Most recent answer

12th Aug, 2018
Zagami Francesco
Tabaka Mission Hospital
For use as an anticoagulant, disodium or tri-potassium salts of EDTA, the optimal concentration is 1.5 mg per ml of blood.

Popular Answers (1)

24th Dec, 2014
Muhammed Malayan
DM Wayanad Institute of Medical Sciences
Potassium EDTA(K2 or K3 ) is more preferred, rather than Sodium EDTA, because Sodium EDTA is less soluble in water.10% solution of potassium EDTA (w/v) in distilled water is prepared as stock anticoagulant for hematological studies. To collect 1ml blood, 10 ul of this solution is added to the collection tube.
24 Recommendations

All Answers (18)

6th Dec, 2014
Anggadia Wardani
Universitas Gadjah Mada
you can add EDTA directly to the collection tube, max about 0,8% of blood sample. make sure the tube is coated properly with EDTA. :D
4 Recommendations
8th Dec, 2014
Vinod Nadella
National Institutes of Health
Hi De,
The K2EDTA prepared as 1% solution in distilled water, and a final concentration of 0.5 ml of anticoagulant for 5 ml blood can be used. It does not dissolve quickly in blood, so the tubes must be inverted multiple times after blood is added. Enough EDTA must be present to prevent coagulation, however, excessive amounts affects both erythrocytes and leukocytes, causing membrane damage. Neutrophils and monocytes appeared to be the cells most sensitive to storage in EDTA, whereas lymphocytes were the most stable.
2 Recommendations
8th Dec, 2014
José Machado da Costa Eduardo
Hospital de São Teotónio
Keep in mind that the usual amount of EDTA needed to avoid blood cloting is something between 1.5 and 1.8 mg per 1mL of blood.
10 Recommendations
17th Dec, 2014
Gisela María Pita-Rodríguez
National Institute of Hygiene, Epidemilogy and Microbiology, Cuba
We use the K2EDTA prepared as 10% solution in distilled water, 10 microliter EDTA for 1ml of blood. This is important in case of can not obtain more than 2ml of total blood for analyses
3 Recommendations
24th Dec, 2014
Muhammed Malayan
DM Wayanad Institute of Medical Sciences
Potassium EDTA(K2 or K3 ) is more preferred, rather than Sodium EDTA, because Sodium EDTA is less soluble in water.10% solution of potassium EDTA (w/v) in distilled water is prepared as stock anticoagulant for hematological studies. To collect 1ml blood, 10 ul of this solution is added to the collection tube.
24 Recommendations
30th Dec, 2014
Morteza Salimian
Kashan University of Medical Sciences and Health Services
There are two options to use EDTA, at first you can add obtained blood to dry EDTA, the concentration of EDTA in Blood would be 1.5mg/mI. However to add EDTA into the tubes you have to transfer a certain volume of the stock EDTA solution5% or 10%,containing the required amount of dry EDTA and then allow to evaporate H20 for overnight. the advantage of this method is lower dilution of blood in anticoagulant. However,You should notice that solubility of dry EDTA is lower compared to EDTA solution. if using dry EDTA its better to choose K3EDTA , Cause its more solubility in H20. Another option is using EDTA solution in blood tubes, in this condition the risk of platelet aggregation declines, but due to dilution of blood sample in Liquid anticoagulant you may have a little decrease in count of blood cells and also Hb . HCT. In this case, K2EDTA is preprefered regarding better preservation of the blood cells morphology.
3 Recommendations
19th Feb, 2016
Omar Faruk Sk Md
Boston Children's Hospital
Can I use this formulation with sheep blood as well....is it possible to use solution of  Na-EDTA dissolved in 0.9% NaCl or PBS?
1 Recommendation
24th Dec, 2016
Md Babu Mia
Icahn School of Medicine at Mount Sinai
When i was preparing Na2EDTA, it got dissolved around pH - 7.4 with NaOH.
3 Recommendations
12th Jan, 2017
Olubukola olusola-makinde
Federal University of Technology, Akure
The disodium salt of EDTA will not go into solution until the pH of the solution is adjusted to ~8.0 by the addition of NaOH (copied)
5 Recommendations
How can we remove plasma from clotted blood?
10th Oct, 2017
Ditthawat Nonghanphithak
Khon Kaen University
Hi Nagendra sastry Yarla
We can separate the plasma from clotted blood by centrifugation.
1 Recommendation
9th Jan, 2018
Zagami Francesco
Tabaka Mission Hospital
Anticoagulant EDTA 0.4 M at pH about 7.4 to 8.0: dissolve 150 g disodium EDTA dihydrate in 700 ml dH2O. Adjust pH to 8.0 adding slowly about 40.3 ml of 10 M NaOH, fill up to 1 liter with dH2O and filter to sterilize. For 10 ml buffered EDTA dissolve 1.5 g of Na2EDTA-2H2O in 5 ml of dH2O, add 0,52 ml of 10 M NaOH (for 10 ml dissolve 4.0 g of NaOH in 5 ml of dH2O. Immerge in cold water until completely dissolved and solution is fresh and fill up to 10 ml with dH2O) and finally fill up to 10 ml with dH2O.
5 Recommendations
7th Apr, 2018
Khalid Al-Salhie
University of Basrah
This link is very useful
Hope you good luck
Regards
1 Recommendation
23rd Apr, 2018
Yesu Raja
Central University of Tamil Nadu
Dear friends...
How long disodium and di potassium EDTA prevent coagulation for human blood?
any exact research paper reports the results..
thanks in advance.
11th May, 2018
Khalid Al-Salhie
University of Basrah
2 Recommendations
11th Aug, 2018
Joseph Yahaya
Ahmadu Bello University
Hi.
Please what amount of EDTA is contained in 5ml EDTA tube?
Help please.
2 Recommendations

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