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Description of key information

The oral LD50 value in rats was between 300 and 2000 mg/kg bw; the 4-h LC50 value of the structurally related substance EDTA-CuNa2 is in excess of 5.30 g/m3. No acute dermal toxicity studies are available but based on read across (see also section 13), acute dermal toxicity following exposure to EDTA-CuK2 is not expected.  

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 September - 24 October 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: Wistar (Han)
Sex:
female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 8-12 weeks old).
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean.
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES: From: 30 September - 24 October 2014
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.
Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 1965 mg/kg body weight

DOSAGE PREPARATION: The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
Doses:
1965 and 300 mg/kg body weight
No. of animals per sex per dose:
9 (3 groups of three females in a stepwise manner)
Control animals:
no
Details on study design:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 1965 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily. Animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons based on OECD guidance document on humane endpoints (ENV/JM/MONO/ 2000/7). The time of death was recorded as precisely as possible.
Body weights: Days 1 (pre-administration), 8 and 15 and at death (if found dead or sacrificed after Day 1).
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: The moribund animal and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
- Other examinations performed: none.
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).
Sex:
female
Dose descriptor:
LD50
Effect level:
300 - 1 965 mg/kg bw
Based on:
test mat.
Mortality:
At 1965 mg/kg, two animals were found dead on Day 1 and 2 and one animal was sacrificed for humane reasons on Day 3. At 300 mg/kg, no mortality occurred.
Clinical signs:
At 1965 mg/kg, lethargy, abnormal posture, hunched posture, uncoordinated movements, piloerection, diarrhoea, pale appearance, ptosis, hypothermia and/or green faeces were noted for the animals between Days 1 and 3. At 300 mg/kg, hunched posture and/or piloerection were noted for the animals between Days 1 and 5.
Body weight:
The body weight gain shown by the surviving animals dosed at 300 mg/kg bw over the study period was considered to be similar to that expected for untreated animals of the same age and strain.
Gross pathology:
At 1965 mg/kg, bluish discouloration of the gastro intestinal-tract was found in all animals, at macroscopic post mortem examination. This was considered to be due to the test substance which was a blue powder. At 300 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 value of EDTA-CuK2 in Wistar rats was established to be within the range of 300-1965 mg/kg body weight.
Executive summary:

The oral LD50 value of EDTA-CuK2 in Wistar rats was established to be within the range of 300-1965 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight. Based on these results:

- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011) (including all amendments), EDTA-CuK2 should be classified as: harmful if swallowed (Category 4) for acute toxicity by the oral route;

- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments), EDTA-CuK2 should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
500 mg/kg bw
Quality of whole database:
One study available with EDTA-CuK2 which showed an LD50 value between 300 and 2000 mg/kg bw; ATE is 500 mg/kg bw. Also one study available with EDTA-Cu(NH4)2 and three studies available with EDTA-CuNa2.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Studies with other metal chelates show low acute inhalation toxicity (see also section 13 for read across).

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Studies with other metal chelates show low acute dermal toxicity (see also section 13 for read across).

Additional information

The oral LD50 value is between 300 and 2000 mg/kg bw. This was also applicable to EDTA-CuK2. Two earlier acute oral toxicity studies were available with EDTA-CuNa2, one indicating an LD50 value of 890 mg/kg bw, the other indicating an LD50 value of ca. 2100 mg/kg bw. The first study mentioned overnight starvation of the rats, the second study did not mentioning anything on this subject. A RF study prior to the start of an extended OECD 422 study with EDTA-CuNa2, using a single exposure study in non-starved rats showed no mortality at a level of 1500 mg/kg bw. It was therefore concluded that there was a very large difference in acute toxicity between starved and non-starved rats.

Justification for selection of acute toxicity – oral endpoint

Relatively well performed and reported study.

Justification for classification or non-classification

EDTA-CuK2 needs to be classified as GHS Cat. 4, harmful if swallowed. No classification needed for acute inhalation toxicity as the 4 -h LC50 of the structurally related substance EDTA-CuNa2 is in excess of 5.30 g/m3. No classification needed for acute dermal toxicity.