Metal ions (calcium, lead, iron, copper, cadmium). No IC50, Ki, EC50, or DC50 values reported. [3]

In vitro experiments showed that Edetate Sodium (EDTA) binds to anion exchange resins (MonoQ, Poros 50 HQ, Q-Sepharose FF, DE52 cellulose, DEAE Sepharose CL-6B) and elutes at NaCl concentrations of approximately 240 mM (for EDTA) at pH 7.9. The binding capacity of MonoQ resin for EDTA was 55.4 ± 0.4 μmol/ml resin (16.2 mg/ml resin). At pH 7.9, EDTA has three negative charges and interacts with positively charged groups on the resin. The measured EDTA capacity was about 2-fold lower than theoretical predictions based on ionic capacity, suggesting that EDTA interacts with only a portion of available charged groups.
A colorimetric chelation assay demonstrated that Edetate Sodium chelates Ni²⁺ ions, causing a change in the visible absorption spectrum of NiCl₂ from 400-800 nm. The presence of EDTA in column fractions was detected by measuring absorbance at 560 nm after mixing with NiCl₂ solution. The concentration of EDTA in the peak fraction was determined to be 22 mM, representing a 220-fold increase from the mobile phase concentration of 0.1 mM.
PCR inhibition assay: Fractions containing high concentrations of Edetate Sodium (22 mM) inhibited Mg²⁺-dependent PCR amplification of the EGFP gene. When 2 μl of each fraction was added to a 25 μl PCR reaction (final volume), only the fraction with the highest EDTA concentration failed to produce the amplified DNA product, confirming that EDTA chelates Mg²⁺ required for Taq DNA polymerase activity.
Protein binding interference: Edetate Sodium binds more tightly to MonoQ resin (eluting at 251 mM NaCl) than EGFP (eluting at 208 mM NaCl). When a MonoQ column was pre-saturated with EDTA, only about 7% of loaded EGFP bound to the resin. Additionally, injection of 5 mM EDTA displaced bound EGFP from the column, with 95% of EGFP eluting as the resin became saturated with EDTA. [1]

Edetate Sodium chelation therapy in patients with claudication (Olszewer, 1990, double-blind, n=10 men): 20 chelation treatments plus magnesium sulfate, B complex, and vitamin C significantly improved walking test (improvement ratio 2.65 vs 1.1 for placebo), Master Exercise Test (1.89 vs 1.02), bicycle stress test (2.10 vs 1.01), resting ankle-brachial index (ABI) (0.89 vs 0.63), and exercise ABI (0.78 vs 0.54) after 10 treatments. After 20 chelation treatments (all subjects received EDTA), mean improvement ratio was 2.93 and ABI at rest 0.95, after exercise 0.88. [3]
Edetate Sodium chelation therapy in patients with claudication (Guldager, 1992, double-blind RCT, n=153): 20 treatments of chelation (3 g) in normal saline with daily oral multivitamins vs placebo. No differences in walking distance or ABI between groups. A subset of 30 subjects with angiograms and transcutaneous oxygen concentrations also showed no improvement. [3]
Edetate Sodium chelation therapy in patients with claudication (Van Rij, 1994, double-blind RCT, n=32): 20 treatments of chelation (3 g in 500 mL normal saline) with daily oral multivitamins vs placebo, twice weekly for 10 weeks. Resting ABI was improved after 3 months in the chelation group, but no other differences in walking distance or postexercise ABIs between groups. [3]
Edetate Sodium chelation therapy in patients with coronary artery disease (Knudtson, 2002, PATCH trial, double-blind RCT, n=84): Chelation (n=41) vs placebo (n=43), twice weekly for 15 weeks and once monthly for an additional 3 months. No difference in exercise time to ischemia between groups after 27 weeks and 33 treatments. A substudy (Anderson, 2003) measured endothelium-dependent brachial artery flow-mediated vasodilation in 23 chelation and 24 placebo subjects at baseline, after first chelation, and at end of study (33 treatments) and found no difference. [3]
Edetate Sodium chelation therapy in post-myocardial infarction patients (Lamas, 2014, TACT trial, multicenter double-blind RCT, n=1708): Patients aged ≥50 years with MI ≥6 weeks before enrollment received 40 intravenous infusions over 28 months (first 30 weekly, then 10 maintenance every 2-8 weeks). Each chelation infusion contained 3 g of Edetate Sodium. Primary endpoint (death, MI, stroke, coronary revascularization, hospitalization for angina) occurred in 139 (32%) of double-placebo group and 108 (26%) of chelation plus vitamin group (HR 0.82; 95% CI 0.69-0.99; P=0.035; NNT over 5 years = 18). In diabetics (n=633), event rate was lower with chelation (HR 0.61; 95% CI 0.45-0.83; P=0.002); diabetics receiving chelation plus vitamins had HR 0.49 (95% CI 0.33-0.75; P<0.001; NNT over 5 years = 5.5). In subjects with anterior wall MI, risk reduction HR 0.63 (95% CI 0.47-0.86; P=0.003). Diabetics treated with chelation experienced 41% relative reduction in combined CVD endpoints (P<0.001), 40% reduction in CVD mortality/nonfatal stroke/nonfatal MI (P=0.017), 52% reduction in recurrent MI (P=0.015), and 43% reduction in all-cause death (P=0.011). [3]

Colorimetric chelation assay for EDTA: A quick colorimetric chelation assay was implemented exploiting EDTA's ability to chelate metal ions. A colored divalent cation, Ni²⁺, was chosen. Visible absorption spectra (400-800 nm) of 0.5 M NiCl₂ were measured in the presence and absence of 0.1 M EDTA. After column chromatography, fractions were collected, lyophilized, solubilized in 100 μl water, and mixed with 100 μl of 1 M NiCl₂. Absorbance at 560 nm was measured to determine the relative concentration of EDTA in each fraction. A standard curve of EDTA concentration vs. absorbance at 560 nm was used to determine the exact concentration. [1]

PCR inhibition assay: PCR was set up using a DNA polymerase kit without MgCl₂. MgCl₂ was added to a final concentration of 1.5 mM prior to amplification. Then 2 μl of each column fraction (eluted during anion exchange chromatography) was added to separate PCR tubes (25 μl total reaction volume). The PCR was used to amplify the full-length sequence of enhanced green fluorescent protein (EGFP) from a plasmid using primers corresponding to the T7 promoter and terminator regions. After 30 cycles of PCR, the amplified DNA was assayed by electrophoresis on a 1% agarose gel. [1]

For the Trial to Assess Chelation Therapy (TACT): Patients aged ≥50 years with prior myocardial infarction (≥6 weeks before enrollment) were randomized to receive Edetate Sodium or placebo infusions. Edetate Sodium was administered as 40 intravenous infusions over 28 months. The first 30 infusions were given weekly, followed by 10 maintenance infusions every 2 to 8 weeks. Each chelation infusion contained 3 g of Edetate Sodium. Patients were further assigned to receive either oral multivitamins and minerals or placebo vitamins. [3]
For the PATCH trial: Patients with diagnosed coronary artery disease received Edetate Sodium chelation therapy twice weekly for 15 weeks and once monthly for an additional 3 months (total 33 treatments). [3]
For claudication trials (Olszewer, Guldager, Van Rij): Edetate Sodium was administered as 20 intravenous infusions. In some protocols, 3 g of Edetate Sodium was given in normal saline, with or without daily oral multivitamins. Treatment frequency was twice weekly for 10 weeks or over 5-9 weeks. [3]

Edetate Sodium has a short lifespan and is eliminated in 12-14 hours. It chelates the large pool of freely diffusible calcium ions in plasma and also chelates free iron in the blood without affecting coagulation function. [5]

Interactions
In humans, the absorption of milk is inhibited when tetracycline hydrochloride, sodium EDTA, and milk are taken simultaneously; however, this inhibition is counteracted by the concurrent intake of EDTA.
Administering cortisol 1-6 hours before EDTA injection reduces the degree of vascular damage in adrenalectomized rats; however, administration of cortisol delayed until 15 minutes before EDTA injection has no protective effect.
Non-human toxicity values Mouse intraperitoneal LD50: 330 mg/kg
Rat intraperitoneal LD50: >2.0 g/kg
Rat intraperitoneal LD50: 4000 mg/kg body weight

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Edetate Sodium chelation therapy is generally well tolerated; the most common side effect is a burning sensation at the site of administration. More serious and potentially fatal side effects occur rarely but include heart failure, hypotension, hypocalcemia, kidney damage, and bone marrow suppression. Hypocalcemia and death may occur particularly if the chelation therapy is infused too rapidly. [3]

[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.

Tetrasodium ethylenediaminetetraacetate (EDTA) is a chelating agent that chelates various polyvalent cations, such as calcium. It is used in pharmaceutical manufacturing and as a food additive.
Therapeutic Uses
When used as a powder chelating agent, tetrasodium EDTA should not come into direct contact with the eyes unless neutralized first, as the resulting solution is alkaline enough to damage the eyes.
Researchers tested the efficacy of tetrasodium EDTA in removing biofilms formed from salivary inoculum or pure cultures of Candida albicans on polymethyl methacrylate (PMMA) denture base discs or toothbrushes used normally for 4–8 weeks. Its virus neutralization efficiency was also determined. After overnight (16 hours) treatment with a 4% (w/v) tetrasodium EDTA solution, the number of viable bacteria in salivary and Candida albicans biofilms decreased by ≥99%. Biofilm removal was confirmed using confocal laser scanning microscopy. The presence or absence of sucrose during biofilm formation had no effect on the bactericidal effect. Prolonged treatment of polymethyl methacrylate (PMMA) with tetrasodium EDTA does not affect the subsequent formation of Candida albicans biofilms or the surface roughness of PMMA, but it does reduce the subsequent formation of biofilms from salivary inoculum. The infectivity of herpes simplex virus and poliovirus suspensions was reduced by more than 99.99% after 1 hour and 2 hours of treatment, respectively. Tetrasodium EDTA solution effectively disinfects toothbrushes and PMMA discs, causing biofilm detachment and rapidly neutralizing both non-enveloped and enveloped viruses. Dentures and toothbrushes are susceptible to contamination by bacterial biofilms and viruses. Therefore, rapid, effective, economical, non-toxic, and easy-to-implement disinfection methods are needed. These studies demonstrate the disinfection value of tetrasodium EDTA solution in oral care. Central venous catheter (CVC)-associated bloodstream infections (BSIs) are known to increase morbidity and mortality in hospitalized and outpatient patients, including those with hematologic malignancies and those undergoing hemodialysis or home infusion therapy. Biofilm-associated microorganisms on the lumen of these catheters have reduced susceptibility to antimicrobial agents. This study tested the bactericidal effect of tetrasodium EDTA as a catheter locking solution on several clinically relevant microbial biofilms. In a model system, biofilms of Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, and Candida albicans were cultured to approximately 1 × 10⁵ CFU/cm², followed by locking with tetrasodium EDTA. Changes in biofilms before and after locking with 40 mg/mL tetrasodium EDTA for 21 hours were compared. The results showed that the viable bacterial counts of all tested microorganisms were significantly reduced after locking (P < 0.05). Antimicrobial locking with 40 mg/mL⁻¹ tetrasodium EDTA for at least 21 hours can significantly reduce and potentially eradicate central venous catheter (CVC)-associated clinically relevant microbial biofilms.
Experimental Applications: Intravenous injection of 50 mg/kg Trilon B into rabbits with lithium chloride poisoning and intraperitoneal injection of 150-175 mg/kg Trilon B into mice both showed significant therapeutic effects.
...Tetrasodium EDTA can effectively treat corneal lime burns and can also be used as a chelating agent to treat hypercalcemia by removing calcium from the blood.

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Edetate Sodium chelation therapy has not been approved by the U.S. Food and Drug Administration (FDA) for the treatment of cardiovascular diseases. Its effectiveness is unclear. [3]
The use of Edetate Sodium chelation therapy to treat CVD and other diseases increased 68% from 2002 to 2007 in the United States according to the 2008 National Health Interview and Nutrition Survey, and an estimated 111,000 individuals are treated yearly with this technique. [3]
Proposed mechanisms of Edetate Sodium in CVD include: binding to calcium to reduce atherosclerotic calcification of arteries, free radical scavenging, reduction of total body iron, cell membrane stabilization, arterial dilatation due to reduced calcium channel activity, improvement of arterial wall elasticity, increased production of nitric oxide, and reduction in lead and cadmium levels. [3]
The TACT investigators concluded that their results did not support the routine use of chelation therapy for post-myocardial infarction patients, despite showing benefit. The TACT consent form did not state that chelation therapy for CVD is not FDA-approved. 18% of TACT subjects were lost to follow-up, and the NHLBI and NCCAM were intentionally unblinded during the study. [3]
A 2002 Cochrane Review concluded that there was insufficient evidence to recommend for or against the use of chelation therapy in CVD management. [3]

380.018

64-02-8

Ethylenediaminetetraacetic acid;60-00-4

6144

White to off-white solid powder

6.9 g/cm3

614.2ºC at 760 mmHg

>300 °C(lit.)

325.2ºC

0

10

7

24

293

0

C(=O)(CN(CCN(CC(=O)[O-])CC(=O)[O-])CC([O-])=O)[O-].[Na+4]

UEUXEKPTXMALOB-UHFFFAOYSA-J

InChI=1S/C10H16N2O8.4Na/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;4*+1/p-4

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

Tetrine Edetate Sodium Trilon B

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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