Copper Chelate Complex

Endpoint summary

Description of key information

Acute oral toxicity:

The single-dose oral toxicity study in rats was performed according to the acute toxic class method (OECD 423 and EU B.1.Tris) in Wistar rats. Two groups of 3 females were treated with the test item at dose level of 300 mg/kg body weight (bw) and one group of 3 females was treated at the dose level of 2000 mg/kg bw. A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. The test item was formulated in DMSO (dimethyl sulfoxide) at concentrations of 30 and 200 mg/mL at a dose volume of 10 mL/kg bw, respectively.

Initially three females (Group 1) were treated at a dose level of 300 mg/kg bw. Only one animal died, therefore the second group of animals (Group 2) were treated at a dose level of 300 mg/kg bw. No mortality was observed, therefore the third group of animals (Group 3) were treated at a dose level of 2000 mg/kg bw. All the three animals died, therefore no further testing was required according to the test guidelines.

At the dose level of 300 mg/kg bw one animal died on Day 1. At the dose level of 2000 mg/kg bw all the animals died on Day 0 (2 hours postdose). After treatment at 300 mg/kg bw, slight to moderate decreased activity (6/6 animals), hunched back (6/6 animals), rooting of bedding (6/6 animals), slight or moderate ataxia (3/6 animals), slight increased salvation (2/6 animals), red discharge (both eyes) (1/6 animal) and piloerection (1/6 animal) were observed. Symptoms were present from Day 0 till Day 2. From Day 3 all animals were symptom-free. After treatment at 2000 mg/kg bw, slight to moderate decreased activity (3/3 animals), hunched back (3/3 animals), rooting of bedding (3/3 animals) and recumbency (3/3 animal) were observed. There were no effects on body weight or body weight gains at dose level of 300 mg/kg bw.

At necropsy, at dose level of 300 mg/kg bw test item-related changes were observed in the stomach and thymus. Blue multifocal discoloration of the stomach glandular mucosa and black diffuse discoloration of the thymus were grossly observed in one found dead female. Other changes were considered to be agonal/post mortem or a common background. At dose level of 2000 mg/kg bw test item-related changes were observed in the stomach, duodenum/jejunum, lungs and trachea. Blue creamy material in the stomach and in the digestive content of duodenum/jejunum, red diffuse discoloration of the stomach glandular mucosa; enlargement, pale/red diffuse/multifocal discoloration of the lungs and white foamy material in the trachea were seen at necropsy.

Under the conditions of this study, the acute oral LD50 value of the test item was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

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:

April - June 2020

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)

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:

no

Specific details on test material used for the study:

- Storage conditions: Controlled room temperature (15-25°C, ≤70% relative humidity), protected from light and humidity (stored in a tightly closed container)

Species:

rat

Strain:

Wistar

Remarks:

strain: Crl:WI

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH (Address: Sandhofer Weg 7, D-97633 Sulzfeld, Germany)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats, 10-11 weeks old
- Weight at study initiation: 205-236 g. The maximum difference of individual animal weights from the mean of the treatment group was not exceed 20%.
- Acclimation period: at least 19 days
- Fasting period before study: night before treatment
- Cage type: Type II. or III. polycarbonate
- Bedding / nesting: Certified laboratory wood bedding and nest building material
- Housing: Group caging (3 animals/cage)
- Diet (e.g. ad libitum): ssniff SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH (Address: D-59494 Soest, Germany) ad libitum.
- Water (e.g. ad libitum): tap water from the municipal supply, as for human consumption from a 500 mL bottle, ad libitum

ENVIRONMENTAL CONDITIONS
- Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
- Temperature: 18.2 – 24.8°C
- Relative humidity: 24 - 68%
- Ventilation: 15-20 air exchanges/hour

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

The test item was formulated in DMSO (dimethyl sulfoxide) at concentrations of 30 and 200 mg/mL at a dose volume of 10 mL/kg bw, respectively.

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

Two groups of 3 female Crl:WI rats were treated with the test item at dose level of 300 mg/kg body weight (bw) and one group of 3 females was treated at the dose level of 2000 mg/kg bw.

Control animals:

no

Details on study design:

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was formulated in DMSO (dimethyl sulfoxide) at concentrations of 30 and 200 mg/mL at a dose volume of 10 mL/kg bw, respectively.
Initially three females (Group 1) were treated at a dose level of 300 mg/kg bw. Only one animal died, therefore the second group of animals (Group 2) were treated at a dose level of 300 mg/kg bw. No mortality was observed, therefore the third group of animals (Group 3) were treated at a dose level of 2000 mg/kg bw. All the three animals died, therefore no further testing was required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris).
Clinical observations were performed at 11, 13, 16, 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter or until death. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy) and at death. All animals were subjected to a necropsy and a macroscopic examination.

Statistics:

At the Test Facility, the statistical evaluation of data was performed using the statistical program package of SAS 9.2 (when using Provantis).

Preliminary study:

None

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

300 - 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At the dose level of 300 mg/kg bw one animal (1/6) died on Day 1. At the dose level of 2000 mg/kg bw all the animals died on Day 0 (2 hours postdose).

Clinical signs:

other: After treatment at the dose level of 300 mg/kg bw, slight to moderate decreased activity (6/6 animals), hunched back (6/6 animals), rooting of bedding (6/6 animals), slight to moderate ataxia (3/6 animals), slight increased salvation (2/6 animals), red di

Gross pathology:

Test item-related macroscopic findings were observed at dose levels of 300 and 2000 mg/kg bw. At dose level of 300 mg/kg bw test item-related changes were observed in the stomach and thymus. Blue multifocal discoloration of the stomach glandular mucosa and black diffuse discoloration of the thymus were grossly observed in one found dead female. Other changes were considered to be agonal/post mortem or a common background.
At dose level of 2000 mg/kg bw test item-related changes were observed in the stomach, duodenum/jejunum, lungs and trachea. Blue creamy material in the stomach and in the digestive content of duodenum/jejunum, red diffuse discoloration of the stomach glandular mucosa; enlargement, pale/red diffuse/multifocal discoloration of the lungs and white foamy material in the trachea were seen at necropsy

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral LD50 value of the test item was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

Executive summary:

The single-dose oral toxicity study in rats was performed according to the acute toxic class method (OECD 423 and EU B.1.Tris) in Crl:WI Wistar rats. Two groups of 3 female Crl:WI rats were treated with the test item at dose level of 300 mg/kg body weight (bw) and one group of 3 females was treated at the dose level of 2000 mg/kg bw. A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. The test item was formulated in DMSO (dimethyl sulfoxide) at concentrations of 30 and 200 mg/mL at a dose volume of 10 mL/kg bw, respectively.

Initially three females (Group 1) were treated at a dose level of 300 mg/kg bw. Only one animal died, therefore the second group of animals (Group 2) were treated at a dose level of 300 mg/kg bw. No mortality was observed, therefore the third group of animals (Group 3) were treated at a dose level of 2000 mg/kg bw. All the three animals died, therefore no further testing was required according to the test guidelines. At the dose level of 300 mg/kg bw one animal died on Day 1. At the dose level of 2000 mg/kg bw all the animals died on Day 0 (2 hours postdose). After treatment at 300 mg/kg bw, slight to moderate decreased activity (6/6 animals), hunched back (6/6 animals), rooting of bedding (6/6 animals), slight or moderate ataxia (3/6 animals), slight increased salvation (2/6 animals), red discharge (both eyes) (1/6 animal) and piloerection (1/6 animal) were observed. Symptoms were present from Day 0 till Day 2. From Day 3 all animals were symptom-free. After treatment at 2000 mg/kg bw, slight to moderate decreased activity (3/3 animals), hunched back (3/3 animals), rooting of bedding (3/3 animals) and recumbency (3/3 animal) were observed. There were no effects on body weight or body weight gains at dose level of 300 mg/kg bw.

At necropsy, at dose level of 300 mg/kg bw test item-related changes were observed in the stomach and thymus. Blue multifocal discoloration of the stomach glandular mucosa and black diffuse discoloration of the thymus were grossly observed in one found dead female. Other changes were considered to be agonal/post mortem or a common background. At dose level of 2000 mg/kg bw test item-related changes were observed in the stomach, duodenum/jejunum, lungs and trachea. Blue creamy material in the stomach and in the digestive content of duodenum/jejunum, red diffuse discoloration of the stomach glandular mucosa; enlargement, pale/red diffuse/multifocal discoloration of the lungs and white foamy material in the trachea were seen at necropsy.

Under the conditions of this study, the acute oral LD50 value of the test item was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

According to the GHS criteria, the test item needs tobe classified as "Toxic, Category 4" for acute oral exposure.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

Klimisch 1, guideline study

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

According to the GHS criteria, the test item needs to be classified as "Toxic, Category 4" for acute oral exposure.