Reaction mass of potassium;sodium;2-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetate;copper and potassium;sodium;2-[2-[bis(carboxymethyl)amino]ethyl-(2-hydroxyethyl)amino]acetate;copper

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

December 17th, 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

dicopper(2+) 2-({2-[bis(carboxymethyl)amino]ethyl}(2-hydroxyethyl)amino)acetate 2-[bis({2-[bis(carboxymethyl)amino]ethyl})amino]acetate

EC Number:

915-008-8

IUPAC Name:

dicopper(2+) 2-({2-[bis(carboxymethyl)amino]ethyl}(2-hydroxyethyl)amino)acetate 2-[bis({2-[bis(carboxymethyl)amino]ethyl})amino]acetate

Test material form:

solid

Remarks:

solid with lumps

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle – France)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 190.0 +- 10.8 g
- Fasting period before study: no
- Housing:
Healthy female rats were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet and water (e.g. ad libitum):
Drinking water (tap-water from public distribution system) and foodstuff (ENVIGO - 2016) were supplied ad libitum. Food was removed on day 1 and then redistributed 4 hours after the test item administration.
Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE). The results of analysis were kept in the Quality Assurance department and then were retained in the Phycher archives as meta-data.
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25°C
- Humidity (%): 30-70%
- Air changes (per hr): at least ten changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (07.00 to 19.00) and twelve hours darkness.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled

Details on oral exposure:

VEHICLE
- Concentration in vehicle:
In the first and second steps of the study, 0.3000 g and 0.3020 g of the test item were weighed and distilled water was added to two 10 mL volumetric flasks. The preparations were stirred by vortex to obtain blue solutions just before the administration.
In the third and fourth steps of the study, 2.0016 g and 2.0004 g of the test item were weighed and distilled water was added to two 10 mL volumetric flasks. The preparations were stirred by vortex to obtain blue solutions just before the administration.
- Amount of vehicle (if gavage): 10 mL/kg body weight
- Justification for choice of vehicle: most suitable formulation at the requested concentrations.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Systematic examinations were carried out to identify any behavioural or toxic effects on the major physiological functions 30 min, 1h, 3h, 4h, 24h, 48h after administration of the test item and continued daily during 14 days.
The animals were weighed on day D0 (just before administering the test item) then on day 2, day 7, and day 14.
- Necropsy of survivors performed: yes. On day 14, the animals were euthanized with Dolethal®. Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.
- Other examinations performed: clinical signs, body weight

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

One mortality was noted in the animals treated at the dose of 2000 mg/kg body weight (1/6), on day 5 during the step 3.

Clinical signs:

other: In the surviving animals (5/6), no clinical signs related to the administration of the test item were observed during the study. No clinical signs related to the administration of the test item were observed before the death.

Gross pathology:

The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.
The macroscopic examination of the dead animal at the end of the study did not reveal treatment related changes.

Other findings:

For the dead animal, rigor mortis and beginning of cannibalism were noted before the necropsy.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of the test item Reaction mixture of CuDTPA and CuHEEDTA is higher than 2000 mg/ kg body weight by oral route in the rat. In accordance with the O.E.C.D. Test Guideline No. 423, the LD50 cut-off of the test item may be considered as 2500 mg/ kg body weight by oral route in the rat.
The test item Reaction mixture of CuDTPA and CuHEEDTA does not have to be classified in accordance with the Regulation EC No. 1272/2008 on classification, labelling and packaging of substances and mixtures.

Executive summary:

The test item Reaction mixture of CuDTPA and CuHEEDTA was administered to a group of 6 female Sprague Dawley rats at the dose of 300 mg/kg body weight and then to a group of 6 female Sprague Dawley rats at the dose of 2000 mg/kg body weight. The experimental protocol was established according to the official method as defined inthe O.E.C.D. Test Guideline No. 423 dated December 17th, 2001 and the test method B.1tris of the Council regulation No. 440/2008.

No mortality was noted in the animals treated at the dose of 300 mg/kg body weight. No clinical signs related to the administration of the test item were observed during the study. The body weight evolution of the animals remained normal during the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

One mortality was noted in the animals treated at the dose of 2000 mg/kg body weight (1/6), on day 5. No clinical signs related to the administration of the test item were observed before the death. An absence of body weight gain was noted on day 2 versus day 0. Then, a decrease of body weight (-23%) was noted on day 5 versus day 0. Rigor mortis and beginning of cannibalism were noted before the necropsy. The macroscopic examination of the animal at the end of the study did not reveal treatment related changes.

In the surviving animals (5/6), no clinical signs related to the administration of the test item were observed during the study. The body weight evolution of the animals remained normal during the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

In conclusion, the LD50of the test itemReaction mixture of CuDTPA and CuHEEDTAis higher than 2000 mg/ kg body weight by oral route in the rat.
In accordance with the O.E.C.D. Test Guideline No. 423, the LD50cut-off of the test item may be considered as 2500 mg/ kg body weight by oral route in the rat.

The test item Reaction mixture of CuDTPA and CuHEEDTA does not have to be classified in accordance with the Regulation EC No. 1272/2008 on classification, labelling and packaging of substances and mixtures.
No signal word or hazard statement is required.