Absorption, Distribution and Excretion
Following oral administration of 180 mg/kg or intravenous injection of 45 mg/kg, rats exhibited relatively high NTA concentrations in the lungs, intestines, and muscles. Distribution patterns depended on the route of administration. Approximately 99% of a single oral dose was eliminated within 24 hours; of this, 96% was excreted in the urine. Following oral administration of NTA-(14)C to rats, 95% was excreted in the urine. Less than 1% was excreted as CO2. NTA absorption from the gastrointestinal tract varied: absorption was higher in dogs than in rats, and higher in rats than in rabbits and monkeys…deposited in the bones. Concentration…increased with increasing frequency of administration. The most active accumulation sites…located in areas of very active bone formation. Although NTA concentrations decreased rapidly after cessation of intake, small amounts of NTA remained in the bones after each administration… In dogs, administration of 10, 20, and 50 mg/kg body weight of hypozinotriacetic acid, respectively, resulted in excretions identified as chondroitin sulfate A and/or chondroitin sulfate C.
In rats where plasma NTA levels reached steady state, the concentration of NTA in the kidneys was higher than that in the plasma. The relatively high NTA concentration in the kidneys can be attributed to the high concentration of NTA in a small amount of urine. NTA is not metabolized in mammals and is rapidly excreted through filtration by the kidneys…
In eight male volunteers who had not taken any medication in the two weeks prior to the start of the study, each was given an oral capsule containing 10 mg of [14C]NTA dissolved in fruit juice. Within 120 hours of administration, 12% of the radioactive material was excreted in the urine as unmetabolized NTA and 77% in the feces. Peak plasma concentrations were reached 12 hours after administration (6.5 ng/g serum)…
Metabolism/Metabolites

Washed cell suspensions of Pseudomonas sp. isolated from wastewater degraded all NTA nitrogen to ammonium before NTA was completely converted to carbon dioxide and water. A small amount of nitrite was also generated. The findings tend to support the argument that NTA degradation occurs via aminodiacetic acid and glycine… In mammalian systems, NTA is not metabolized but is rapidly excreted through filtration by the kidneys.

Interactions
Mice injected subcutaneously with cadmium chloride and simultaneously with NTA showed histological evidence of liver necrosis 24 hours later. In pregnant rats, a single intravenous injection of lead (50 mg/kg) or an equimolar amount of chelating agent increased fetal lead levels by approximately 400% after injection of lead penicillamine lead (PBPEN) and lead hypotriacetate (PBNTA), and by 50% after injection of lead EDTA. Following cannulation with (14)C-labeled NTA (68.2), NTA levels in all tissues were lowest in lead-treated and untreated rats. Clearly, lead promoted NTA clearance. This study investigated the effect of a single intravenous injection of 54Mn(II) on the clearance of radioactive manganese in various organs and plasma of rats. Compared with the control group, the NTA-treated rats showed reduced levels of radioactive manganese, indicating that NTA rapidly binds and forms stable and diffusible complexes, thereby promoting the rapid excretion of injected 54Mn(II). This study also explored the interactions of carcinogen pairs acting on different organ systems in rats, identified common design problems in such studies, and tested novel statistical methods. Fischer-344 rats were dietaryly exposed to six possible combinations of four chemicals. The test chemicals and their target organs were as follows: N-methyl-N'-nitro-N-nitrosoguanidine, stomach; N-butyl-N-(4-hydroxybutyl)nitrosamine, bladder; dipentylnitrosamine, liver; and hypozinotriacetic acid, kidney, ureter, and bladder. A 4×4 factorial design was used to study each chemical combination. The dosages were: N-methyl-N'-nitro-N-nitrosoguanidine 0, 20, 40, or 80 ppm; N-butyl-N-(4-hydroxybutyl)nitrosamine 0, 30, 60, or 120 ppm; dipentylnitrosamine 0, 50, 150, or 450 ppm; and hypozoxytriacetic acid 0, 200, 2000, or 20000 ppm. The study period was 104 weeks. An additive index, indicating the interaction and its type, was calculated using an independent interaction model. The endpoints investigated included the incidence of malignancy, time to all-cause mortality, and time to malignancy mortality. The results showed that dipentylnitrosamine did not interact with the other three carcinogens. The combination of hypozoxytriacetic acid with N-methyl-N'-nitro-N-nitrosoguanidine or N-butyl-N-(4-hydroxybutyl)nitrosamine resulted in antagonistic effects on the target sites of the latter two compounds; however, no interaction was observed between N-methyl-N'-nitro-N-nitrosoguanidine and N-butyl-N-(4-hydroxybutyl)nitrosamine. No synergistic effects were observed in any combination or target site. The conclusion is that antagonistic effects may occur when these compounds are administered in combination to rats; although the combined exposure may be highly carcinogenic, the number of tumors produced is not higher than that predicted by the independent-action model.

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Non-human toxicity values
Oral LD50 in mice: 3160 mg/kg
Oral LD50 in rats: 1470 mg/kg

[1]. A review of the environmental and mammalian toxicology of nitrilotriacetic acid. Crit Rev Toxicol. 1985;15(1):1-102.

According to an independent committee of scientific and health experts, hypotriacetic acid (HTA) may be carcinogenic. Hydrazotriacetic acid is a tasteless, white solid that sinks and mixes with water. (US Coast Guard, 1999) HTA is a tricarboxylic acid and an NTA (hydrazotriacetic acid ester). It is nephrotoxic and carcinogenic. It is the conjugate acid of the hypotriacetic acid ion (1-). HTA is a derivative of acetic acid (N(CH2COOH)3). It is a complexing agent (chelating agent) that forms stable complexes with Zn2+. (From the Miar Chemical Dictionary, 5th edition) HTA is a white crystalline solid compound. HTA is primarily used as a chelating agent and eluent, commonly found in laundry detergents. Exposure to HTA can irritate the skin, eyes, and respiratory tract, and cause kidney and bladder damage. HTA is reasonably expected to be a human carcinogen. (NCI05)
A derivative of acetic acid, N(CH2COOH)3. It is a complexing agent (chelating agent) that can form stable complexes with Zn2+.
A derivative of acetic acid, N(CH2COOH)3. It is a complexing agent (chelating agent) that can form stable complexes with Zn2+. (Excerpt from Miyar Chemical Dictionary, 5th Edition)

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Mechanism of Action
The combined use of hyponitrotriacetic acid (HNATA) and soluble hexavalent chromium (in the form of potassium dichromate) has a synergistic effect on gene mutations in Salmonella typhimurium and Drosophila melanogaster. Studies have shown that this effect may depend on the effect of HNATA on the reduction of hexavalent chromium in cellular proteins. Gene mutations were detected in Salmonella typhimurium strains (TA-100), (TA-92), (TA-104), and (TA-103) using the Ames plate incorporation assay. In both Salmonella and Drosophila systems, hydantoin synergistically enhances the mutagenicity of subtoxic doses of hexavalent chromium; however, at higher doses of hexavalent chromium, the presence of hydantoin leads to a decrease in mutation frequency, likely due to toxicity. Both effects may be related to the increased effectiveness of the final mutagen in the presence of hydantoin. This interaction is particularly evident in strains (TA-100) and (TA-104) carrying mutations affecting cell wall permeability and DNA repair. In these strains, the uptake of hexavalent chromium and hydantoin is increased, resulting in reduced efficiency of DNA damage repair or repair via error-prone mechanisms. Hydantoin may promote the uptake of chromate by cell membrane anion carriers. Other mechanisms related to its chelating effect may also be important; for example, very low doses of EDTA have a significant synergistic effect on the mutagenicity of hexavalent chromium, without affecting the reduction of hexavalent chromium by Salmonella proteins under cell-free conditions.

C6H9NO6

191.1388

191.042

139-13-9

Nitrilotriacetic acid trisodium salt;5064-31-3;Nitrilotriacetic acid-d9;807630-34-8;Nitrilotriacetic acid disodium salt;15467-20-6

8758

White to off-white solid powder

1.6±0.1 g/cm3

498.2±40.0 °C at 760 mmHg

245 °C (dec.)(lit.)

255.1±27.3 °C

0.0±2.7 mmHg at 25°C

1.558

-1.76

3

7

6

13

187

0

MGFYIUFZLHCRTH-UHFFFAOYSA-N

InChI=1S/C6H9NO6/c8-4(9)1-7(2-5(10)11)3-6(12)13/h1-3H2,(H,8,9)(H,10,11)(H,12,13)

2-[bis(carboxymethyl)amino]acetic acid

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.)