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

Endpoint summary

Description of key information

Information on the substance itself was only available on acute oral toxicity (guideline study) and two guideline studies were available on for two category members: one on oral toxicity (Cu chelate) and one on inhalation toxicity (Zn chelate). In all studies, no effects were observed at concentrations up to the maximum dose for classification (2000 mg/kg for oral study, 5 mg/L for inhalation study).

Therefore, it was concluded that the substance itself also does not lead to acute toxic effects.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH

1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
See the documentation in support for the category approach.

2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL
See the documentation in support for the category approach.

Qualifier:

according to guideline

Guideline:

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

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.

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 itemReaction mixture of CuDTPA and CuHEEDTAwas 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 itemReaction mixture of CuDTPA and CuHEEDTA does not have to be classifiedin 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.

Reference 2

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

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 17th, 2001

Deviations:

no

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

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.

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

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.

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH

1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
See the documentation in support for the category approach.

2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL
See the documentation in support for the category approach.

Qualifier:

according to guideline

Guideline:

OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)

Version / remarks:

September 2009

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Han

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult animals, approximately 11 weeks old
- Weight at study initiation: Males: 308 to 326 / Females: 191 to 225 g
- Fasting period before study: no
- Housing:
On arrival and following assignment to the study, animals were group housed (up to 5 animals of the same sex and same exposure group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room(s) in which the animals were kept were documented in the study records.
Animals were separated during designated procedures/activities. Each cage was clearly labeled.
- Diet (e.g. ad libitum):
Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility.
It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum):
Municipal tap-water was freely available to each animal via water bottles.
Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility.
It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period:
The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.

ENVIRONMENTAL CONDITIONS
Target temperatures of 18 to 24°C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 20 to 22°C with an actual daily mean relative humidity of 44 to 53%. A 12 hour light/12 hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

IN-LIFE DATES: From: 25/01/2018 To: 08/02/2018

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

4 µm

Geometric standard deviation (GSD):

>= 1.9 - <= 2

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The design of the exposure chamber is based on the directed flow nose only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983).
- Exposure chamber volume: /
- Method of holding animals in test chamber: polycarbonate restraining tubes
- Source and rate of air: The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min.
- Method of conditioning air: /
- System of generating particulates/aerosols:
Administering the test item to a stream of pressurized air using a combination of a spiral feeder (Randcastle Extrusion Systems, Cedar Grove, NJ, USA) with a vibrator attached and air mover (AIR-VAC, Milford, CT, USA) generated an aerosol. The aerosol was passed through a series of two cyclones, allowing larger particles to settle. The primary aerosol was diluted with pressurized air before it entered the exposure chamber (Figure 1.1). The mean total airflow was 17 L/min.
From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.
- Method of particle size determination:
The particle size distribution was characterized twice during each exposure period. The samples were drawn with a flow of 2 L/min. from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber (Figure 1.2). The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (TE-290-GF. Tisch Environmental, Cleves, Ohio, USA) and a fiber glass back-up filter (SEC-290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test item collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined based on OECD guidance document No 39. Graphs of the cumulative mass of test item collected (percentage of total collected) against the cut points of the impactor stages were drawn on log-normal paper. When drawing the graphs more weight was given to the cut points where the cumulative mass sampled was within the range of 5 to 95%. The Mass Median Aerodynamic Diameter (MMAD), i.e. the particle size where 50% of the particle mass was borne by particles smaller than the MMAD and the PS84, (the particle size where 84% of the particle mass was borne by particles smaller than the PS84 was read from the graph. The geometric standard deviation (gsd) was calculated as PS84 / MMAD.

- Temperature, humidity, pressure in air chamber:
The temperature and relative humidity were measured with a humidity and temperature indicator (E+E Elektronik, Engerwitzdorf, Austria) and recorded after the animals were connected to the exposure chamber and at 30 minute intervals after initiation of exposure. The probe was inserted in a tube mounted in one of the free animal ports of the exposure chamber (Figure 1.2). The temperature of the atmosphere during the exposure was between 20.7 and 21.7oC. The relative humidity was between 18 and 21% which was considered appropriate for this relatively short 4 hours exposure duration.

TEST ATMOSPHERE
- Brief description of analytical method used:
A total of 20 representative samples were taken for determination of the actual concentration during exposure at 5 mg/L. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber. Samples were drawn through a glass fiber filter (type APFC04700, Millipore, Billerica, MA, USA). Sample volumes were measured by means of a dry gas meter (type G 1.6, Actaris Meterfabriek B.V., Dordrecht, The Netherlands). The collected amount of test item in the air sample was measured gravimetrically. Subsequently the time-weighted mean concentration with the standard deviation was calculated.
- Samples taken from breathing zone: yes

VEHICLE
- Composition of vehicle (if applicable): air
- Concentration of test material in vehicle (if applicable): 5 mg/L
- Justification of choice of vehicle: standard protocol
- Lot/batch no. (if required): /
- Purity: /

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5 mg/L

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.
Animals were checked for mortality, behavioral signs of distress and effects on respiration at least three times during exposure.
Post exposure observations were performed at periodic intervals on the day of exposure (at least two times) and once daily thereafter. The observation period was 14 days.
Animals were weighed individually on Day 1 (pre exposure), 2, 4 and 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs (animals were checked for mortality, behavioral signs of distress and effects on respiration), body weight,

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.1 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred

Clinical signs:

other: During exposure, no clinical signs were seen (not presented in the table). After exposure, hunched posture was seen for all animals 1 hour after exposure. The animals had recovered from the clinical signs within 3 hours after exposure.

Body weight:

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The inhalation LC50, 4h value of Reaction mixture of ZnEDTA, ZnDTPA and ZnHEEDTA in Wistar rats was established to exceed 5 mg/L.
Based on these results Reaction mixture of ZnEDTA, ZnDTPA and ZnHEEDTA does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Executive summary:

The objective of this study was to assess the acute inhalation toxicity of Reaction mixture of ZnEDTA, ZnDTPA and ZnHEEDTA in rats of both sexes in rats following a single 4 hour nose-only exposure to one or more defined concentrations. Animals were retained for a 14 day post exposure observation period.

 

The study was carried out based on the guidelines described in:

·      Acute Inhalation Toxicity- Acute Toxic Class Method, September 2009.

 

Reaction mixture of ZnEDTA, ZnDTPA and ZnHEEDTA was administered as a dust by nose only inhalation for 4 hours to one group of three male and three female Wistar rats at a target concentration of 5 mg/L. Mortality and clinical signs were observed daily during the observation period and body weights were determined on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (Day 15).

 

The time-weighted mean actual concentration was 5.1 ± 0.13 mg/L. The nominal concentration was 203 mg/L. The generation efficiency was 2.5%.

The mass median aerodynamic diameter (MMAD) was 4.0 µm and 4.0 µm with a geometric standard deviation (gsd) of 1.9 and 2.0, respectively. 

No mortality occurred.

During exposure, no clinical signs were seen. After exposure, hunched posture was seen for all animals 1 hour after exposure. The animals had recovered from the clinical signs within 3 hours after exposure. 

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

No abnormalities were found at macroscopic post mortem examination of the animals.

 

The inhalation LC_{50, 4h}value of Reaction mixture of ZnEDTA, ZnDTPA and ZnHEEDTA in Wistar rats was established to exceed 5 mg/L.

Based on these results Reaction mixture of ZnEDTA, ZnDTPA and ZnHEEDTA does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because skin contact in production and/or use is not likely

the study does not need to be conducted because inhalation of the substance is likely

Endpoint conclusion

Endpoint conclusion:

no study available