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

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:

trimanganese(2+) hexasodium 2-({2-[bis(carboxylatomethyl)amino]ethyl}(2-hydroxyethyl)amino)acetate 2-({2-[bis(carboxylatomethyl)amino]ethyl}(carboxylatomethyl)amino)acetate 2-[bis({2-[bis(carboxylatomethyl)amino]ethyl})amino]acetate potassium hydride sodium hydride

EC Number:

915-009-3

IUPAC Name:

trimanganese(2+) hexasodium 2-({2-[bis(carboxylatomethyl)amino]ethyl}(2-hydroxyethyl)amino)acetate 2-({2-[bis(carboxylatomethyl)amino]ethyl}(carboxylatomethyl)amino)acetate 2-[bis({2-[bis(carboxylatomethyl)amino]ethyl})amino]acetate potassium hydride sodium hydride

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: 194.7 +- 9.2 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 step of the study, 1.018 g and 1.0008 g of the test item were weighed and distilled water was added to two 5 mL volumetric flasks. The preparations were stirred by vortex to obtain yellowish solutions then pooled just before the administration.
In the second step of the study, 2.0023 g of the test item were weighed and distilled water was added to a 10 mL volumetric flask. The preparation was stirred by vortex to obtain a yellowish solution 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:

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:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs related to the administration of the test item were observed during the study.

Gross pathology:

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

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of the test item Reaction mixture of MnEDTA, MnDTPA and MnHEEDTA 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 to be higher than 5000 mg/ kg body weight by oral route in the rat.
The test item Reaction mixture of MnEDTA, MnDTPA and MnHEEDTA 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.

Executive summary:

The test item Reaction mixture of MnEDTA, MnDTPA and MnHEEDTA was administered, 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, 2001andthe test method B.1tris of the Council Regulation No. 440/2008.

No mortality occurred during the study.
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 LD50 of the test item Reaction mixture of MnEDTA, MnDTPA and MnHEEDTA 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 LD50cut-off of the test item may be considered to be higher than 5000 mg/ kg body weight by oral route in the rat.

The test item Reaction mixture of MnEDTA, MnDTPA and MnHEEDTA 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.