Ethylenediaminetetraacetic acid trisodium salt (EDTA) at high concentrations eluted from anion exchange columns can inhibit Mg²⁺-dependent polymerase chain reaction (PCR). When 2 µl of a fraction containing an enriched concentration of EDTA (22 mM, representing a 220-fold increase from the original 0.1 mM in the mobile phase) was added to a PCR reaction, it completely inhibited the amplification of the EGFP gene, unlike fractions without the EDTA peak. [1]
EDTA present in protein purification buffers can interfere with anion exchange chromatography (AEC). When an AEC column (e.g., MonoQ) was equilibrated and samples were applied in buffers containing EDTA, the EDTA bound to the resin and co-eluted as a sharp peak during a salt gradient. This concentrated EDTA peak (eluting at ~240-275 mM NaCl depending on resin and pH) can mask protein peaks detected at 215 nm, displace weakly bound proteins (e.g., EGFP), and reduce the protein-binding capacity of the resin. Saturation of the resin with EDTA (e.g., by applying 5 mM EDTA) can prevent subsequent binding of proteins like EGFP, with only ~7% of the protein adsorbing under such conditions. Conversely, applying a high concentration of EDTA (5 mM) to a column with pre-bound EGFP can displace approximately 95% of the protein. [1]
As an in vitro anticoagulant, Ethylenediaminetetraacetic acid trisodium salt (EDTA) prevents blood clotting by chelating calcium ions, which are necessary for the coagulation cascade. It is the recommended anticoagulant for hematological testing (complete blood count) as it optimally preserves cellular components and blood cell morphology. [2]
EDTA demonstrates a stabilizing effect on labile molecules in blood samples. For example, EDTA plasma is recommended for the measurement of hormones like corticotropin (ACTH), parathyroid hormone (PTH), glucagon, C-peptide, and others, as it inhibits degradation. PTH is more stable in EDTA plasma than in serum. [2]
EDTA can inhibit complement activation and stabilize certain proteins in immunoassays when added to the reaction buffer. [2]
EDTA is the anticoagulant of choice for molecular biology applications (e.g., PCR, HIV-1 RNA quantification) as it stabilizes nucleic acids and, unlike heparin, does not inhibit DNA amplification. HCV-RNA concentrations in EDTA anticoagulated blood are stable for up to 5 days at room temperature. [2]
EDTA can cause interference in specific diagnostic assays: It can lead to underestimation of Troponin T (TnT) by up to 18% in some immunoassays and can interfere with Troponin I (TnI) activity by chelating Ca²⁺ needed for the complex. EDTA is not recommended for their measurement. [2]
EDTA may cause a variable degree of interference in some myoglobin immunoassays. [2]
For natriuretic peptides (ANP, BNP), EDTA plasma (with or without antiproteolytic substances) is recommended for collection, and cold storage in EDTA tubes is indicated. However, NT-proBNP concentration in EDTA plasma can be up to 10% lower than in serum or heparin plasma. [2]
EDTA is recommended for proteomics studies as its chelating action inhibits metal-dependent proteases, helping to stabilize the protein profile. [2]
EDTA is recommended for measuring certain drugs like aminoglycosides and some antiepileptic drugs. It can serve as an alternative to heparin or serum for measuring several antiepileptic drugs, antiarrhythmic drugs, salicylate, acetaminophen, and theophylline. However, it should not be used for measuring 3,4-dihydroxyphenylalanine (DOPA), which undergoes rapid degradation triggered by EDTA. [2]
In clinical chemistry, EDTA is traditionally unsuitable for general analyses due to its chelation of metal ions (e.g., Ca, Mg, Fe). However, it is the anticoagulant of choice for specific tests like ammonia, creatine kinase (CK) isoforms (where zinc chelation is crucial), and homocysteine assays. EDTA plasma can also be used for measuring total cholesterol, HDL-C, LDL-C, and apolipoproteins, though concentrations may be underestimated compared to serum depending on the analytical technique. [2]
EDTA can induce several artifacts: It can cause time-dependent platelet shape change (discoidal to spherical), leading to an increase in Mean Platelet Volume (MPV) measured by impedance analyzers, necessitating measurement at a fixed time after blood draw. [2]
EDTA can induce pseudothrombocytopenia, a phenomenon characterized by in vitro platelet clumping or adhesion to white blood cells, leading to spuriously low platelet counts. This is often mediated by IgM autoantibodies against platelet glycoproteins IIb/IIIa, whose conformation is altered by EDTA. The prevalence is nearly 0.1% in the general population. [2]
EDTA has been associated with other rare phenomena such as leukocyte clumping, erythrocyte agglutination, and pseudoleukocytosis. [2]
EDTA inhibits lipopolysaccharide (LPS)-induced cytokine production (e.g., TNF-α) in whole blood assays, making it unsuitable for measuring TNF-α in such contexts. [2]

[1]. Artifact-inducing enrichment of ethylenediaminetetraacetic acid and ethyleneglycoltetraacetic acid on anion exchange resins. Anal Biochem. 2011 May 1;412(1):34-9.

[2]. The role of ethylenediamine tetraacetic acid (EDTA) as in vitro anticoagulant for diagnostic purposes. Clin Chem Lab Med. 2007;45(5):565-76.

[3]. Chelation therapy in the treatment of cardiovascular diseases. J Clin Lipidol. 2016 Jan-Feb;10(1):58-62.

[4]. The effect of ethylenediaminetetra-acetic acid on the cell walls of some gram-negative bacteria. J Gen Microbiol. 1965 Jun;39(3):385-99.

[5]. Ethylenediaminetetraacetic acid induces antioxidant and anti-inflammatory activities in experimental liver fibrosis. Redox Rep. 2011;16(2):62-70.

[6]. Remediation of heavy metals contaminated silty clay loam soil by column extraction with ethylenediaminetetraacetic acid and nitrilo triacetic acid. Journal of Environmental Engineering, 2017, 143(8): 04017026.

[7]. Ethylenediaminetetraacetic acid (EDTA) enhances cAMP production in human TDAG8-expressing cells. Biochem Biophys Res Commun. 2022 Oct 20;626:15-20.

Trisodium ethylenediaminetetraacetate is an odorless white crystalline powder. The pH value (1% aqueous solution) is 9.3 and the pH value (10% aqueous solution) is approximately 8.3-8.7. (NTP, 1992)
Trisodium ethylenediaminetetraacetate is an organic sodium salt of ethylenediaminetetraacetic acid (EDTA), in which three sodium ions are bound to one EDTA trianion. It contains one EDTA(3-) ion.

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Trisodium ethylenediaminetetraacetate (EDTA) is widely used as an additive in protein purification buffers. Its functions include chelating divalent and trivalent cations to reduce the activity of metal ion-dependent proteases, reducing metal ion-catalyzed protein oxidative damage, stabilizing reducing agents (such as DTT), and reducing the formation of disulfide bonds. [1] EDTA is a polyvalent anion whose charge varies with pH (the pKa values of its carboxylic acid are 2.0, 2.7, 6.2 and 10.3, respectively). It has a strong interaction with anion exchange resins, including strong “Q” and weak “DEAE” types. When EDTA is present in the mobile phase, it is adsorbed and enriched on these resins during column equilibration and sample loading. At pH 7.9, when the NaCl concentration is about 205-254 mM (depending on the resin), EDTA will elute as a distinct peak in salt gradient elution. The on-column enrichment effect can make the concentration of EDTA in the eluent 10 to 220 times higher than its initial concentration in the injection or buffer. [1] This enrichment effect can cause a number of artifacts in protein purification and downstream applications: it reduces the binding capacity of the resin to proteins, displaces weakly bound proteins, masks protein peaks when monitored at 215 nm absorbance, causes an abnormal drop in pH after the addition of Ca²⁺, and inhibits metal-dependent detection (such as PCR). The components co-eluted with the EDTA peak may contain “mystery inhibitors”. [1] To mitigate these effects, the literature recommends: equilibrating the AEC column in an EDTA-free mobile phase and switching to an EDTA-containing buffer before loading; using a low concentration of EDTA (0.1-0.5 mM) in the mobile phase; monitoring elution at 280 nm and 215 nm; adding EDTA to the fraction eluted before the EDTA peak to the desired concentration; and using a control experiment with a buffer-only sample to determine the position and concentration of the EDTA peak. In the final purification step, EDTA can be omitted from the mobile phase and added directly to the fraction collected after elution. [1] Trisodium ethylenediaminetetraacetate (EDTA) is a polybasic acid containing four carboxylic acid groups and two amino groups, which can chelate metal ions. In diagnostic applications, it is mainly used as an in vitro anticoagulant. [2] Commonly used EDTA salts include Na₂EDTA, K₂EDTA, and K₃EDTA. The International Committee for Standardization of Hematology (ICSH) recommends K₂EDTA as the preferred anticoagulant for hematological tests. K₂EDTA causes less cell contraction than K₃EDTA. The recommended concentration is 1.5 mg/mL blood (or 4.55 mmol/L). [2] EDTA tubes are usually color-coded and have a light purple stopper. [2] EDTA plasma is not suitable for measuring electrolytes such as calcium, magnesium, iron, sodium, or potassium (because it is used in the form of sodium or potassium salts). [2] Hematological parameters of EDTA-anticoagulated blood are generally stable: hemoglobin is stable for up to 48 hours at 4°C, and erythrocyte parameters are stable for up to 24 hours. Reticulocyte analysis is recommended to be performed within 24 hours. [2] To alleviate pseudothrombocytopenia caused by EDTA, other anticoagulants such as sodium citrate, heparin, or CTAD (citrate, theophylline, adenosine, glucose) can be used for platelet counting. Adding aminoglycoside antibiotics (such as kanamycin) to EDTA samples can prevent and separate in vitro platelet aggregation. [2] For cytokine assays, EDTA is recommended for sample collection and stabilization until centrifugation, which should be performed rapidly. However, EDTA is not recommended for the assay of soluble interleukin-2 receptor (sIL-2R), soluble transferrin receptor (sTfR), or TNF-α in LPS stimulation assays. [2]
This review explores the potential of a "universal anticoagulant" in simplifying laboratory workflows and notes that EDTA is considered for this purpose due to its stability, even for coagulation assays in patients taking oral anticoagulants. However, alternatives such as hirudin and synthetic thrombin inhibitors (e.g., argatroban, PPACK) have also been explored. [2]
The summary table in this review provides the following recommendations: EDTA is recommended for blood cell counts, easily enzymatically degradable proteins/peptides, proteomics, molecular biology, virology, and specific clinical chemistry assays (ammonia, CK isoenzymes). It is not recommended for general clinical chemistry, troponin assays, or metalloproteinase assays. [2]

C10H16N2O8.NA+

315.23244

358.036

150-38-9

EDTA dipotassium dihydrate;25102-12-9;Ethylenediaminetetraacetic acid;60-00-4

9008

White to off-white solid powder

614.2ºC at 760 mmHg

237 °C

325.2ºC

1.15E-16mmHg at 25°C

1

10

8

23

351

0

QZKRHPLGUJDVAR-UHFFFAOYSA-K

InChI=1S/C10H16N2O8.3Na/c13-7(14)3-11(4-8(15)16)1-2-12(5-9(17)18)6-10(19)20;;;/h1-6H2,(H,13,14)(H,15,16)(H,17,18)(H,19,20);;;/q;3*+1/p-3

trisodium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxymethyl)amino]acetate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~50 mg/mL (~139.59 mM)

Solubility in Formulation 1: 50 mg/mL (139.59 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)