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

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.

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.

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

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.

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.

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?

When i was preparing Na2EDTA, it got dissolved around pH - 7.4 with NaOH.

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)

How can we remove plasma from clotted blood?

Hi Nagendra sastry Yarla

We can separate the plasma from clotted blood by centrifugation.

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.

This link is very useful

http://www.protocol-online.org/biology-forums/posts/28673.html

Hope you good luck

Regards

Dear friends...

How long disodium and di potassium EDTA prevent coagulation for human blood?

any exact research paper reports the results..

thanks in advance.

This link is helpful

https://sciencing.com/make-edta-solution-6452756.html

regards

these links are useful

https://www.sigmaaldrich.com/content/dam/sigma.../ed2ppis.pdf

apps.who.int/iris/bitstream/.../1/WHO_DIL_LAB_99.1_REV.2.pdf

https://thl.fi/publications/ehrm/product2/part_iii4.htm

regards

Hi.

Please what amount of EDTA is contained in 5ml EDTA tube?

Help please.

For use as an anticoagulant, disodium or tri-potassium salts of EDTA, the optimal concentration is 1.5 mg per ml of blood.