Copper dioleate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 November - 7 Dec 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (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:

Copper dioleate

EC Number:

214-307-4

EC Name:

Copper dioleate

Cas Number:

1120-44-1

Molecular formula:

C18H34O2.1/2Cu

IUPAC Name:

copper(2+) bis((9Z)-octadec-9-enoate)

Test material form:

solid

Remarks:

paste

Details on test material:

Solid green paste. Batch number 21092015

Specific details on test material used for the study:

Test Item: Copper dioleate
Lot Number: 21092015
CAS No. 1120-44-1
Purity > 99 %, IR
Appearance: Green, solid/paste
Composition: 99-100%
Homogeneity: Homogeneous
Expiry Date: 21.09.2018
Storage Room temperature: (20 ± 5 °C)
Test Item Receipt According to SPPA-00147-BIO, Test and Reference Items

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Test Animals
Species: Wistar rats
Source: Dobrá Voda, Slovak Republic
Number and Sex of Animals:6 females
Age at First Dose: 8-12 weeks; female animals were non-pregnant and nulliparous
Animal Health: The health condition of animals was examined by a veterinarian before initiation of the study.
Acclimation: The animals were acclimated under the conditions identical to the conditions during the experiment 5 days prior to the start of treatment. The acclimation was according to the standard operation procedure.
Housing Condition: The animals were housed in plastic cages suspended on stainless steel racks, up to 3 animals per cage in a room equipped with central air-conditioning. The average room temperature was maintained within the range of 22.1 ± 0.3° C, relative humidity within 56.3 ± 2.5 %. The light regimen was set to a 12-hour light /12-hour dark cycle. The sanitation was performed according to the standard operation procedures.
Diet: A laboratory food ssniff (Spezialdiäten GmbH, Germany) was offered in recommended doses each day approximately at the same time. The certificate of analysis is included in the raw data.
Water: The animals received tap water for human consumption. Supply of drinking was unlimited. The quality of drinking water is periodical analysed (including microbiological control) and recorded; certificate of analysis is included in raw data.
Bedding: Lignocel S3/4, Lufa - ITL GmbH, Germany
Animals Identification: Each animal was marked with an ID number. Each cage was affixed with a cage card containing pertinent animal and study information.
The animals in cages were marked by a line on the tail with an ink marker.
Justification for the Choice of Species: Normally females are used in the test according to OECD TG 423 because mostly females are the more sensitive gender.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Remarks:

Olive oil is a standard vehicle according to OECD TG 423

Details on oral exposure:

Number of Animals and Dose Levels
The starting dose could be selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg body weight. Available information indicated that the test item is likely to be non-toxic with regard to acute toxicity. A limit dose of 2000 mg/kg body weight was used as a starting dose. One group of 3 females was dosed. Test item-related mortality was not observed during 24 hours and therefore in a second step another 3 females were treated at the same dose.

Doses:

Dose Preparation:
The required amount of the test item (according to the body weight and dose) was mixed with vehicle (olive oil) shortly before administration.
Dose Administration:
The test item was administered in a single dose by gavage using a metal stomach tube. Animals were fasted prior to dosing (food but not water were withheld over-night). Following the period of fasting, the animals were weighted, and the test item was administered. After the test item had been administered, food was withheld for further 3-4 hours.

No. of animals per sex per dose:

One group of 3 females was dosed. Test item-related mortality was not observed during 24 hours and therefore in a second step another 3 females were treated at the same dose.

Control animals:

no

Details on study design:

The test item was administered in a single dose by gavage using a metal stomach tube. Animals were fasted prior to dosing (food but not water were withheld over-night). Following the period of fasting, the animals were weighted, and the test item was administered. After the test item had been administered, food was withheld for further 3-4 hours.
Clinical Observation:
The animals were observed individually immediately after the administration of the test item and then 0.5, 1, 2, and 4 hours later. Then each animal was inspected daily for the next 14 days.
Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body Weight:
Individual weights of animals were determined shortly before the test item was administered and weekly thereafter. Weight differences after first and second weeks after administration were calculated and recorded.
Necropsy:
All test animals were subjected to gross necropsy and result were recorded for each animal.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No mortality was observed during the study. All 6/6 females survived the limit dose of 2000 mg/kg body weight.

Clinical signs:

Animals lived through observation period without signs of intoxication. Neither change of health nor negative reactions were registered.
Clinical Observation
Observation Time After Administration
Hour Day
Immediately 0.5 1 2 4 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Skin and Hair - - - - - - - - - - - - - - - - - - -
Eyes - - - - - - - - - - - - - - - - - - -
Mucosa - - - - - - - - - - - - - - - - - - -
Respiratory System - - - - - - - - - - - - - - - - - - -
Circulatory System - - - - - - - - - - - - - - - - - - -
CNS - - - - - - - - - - - - - - - - - - -
Somatomotoric Activity - - - - - - - - - - - - - - - - - - -
Tremor - - - - - - - - - - - - - - - - - - -
Spasms - - - - - - - - - - - - - - - - - - -
Salivation - - - - - - - - - - - - - - - - - - -
Diarrhoea - - - - - - - - - - - - - - - - - - -
Lethargy - - - - - - - - - - - - - - - - - - -
Sleep - - - - - - - - - - - - - - - - - - -
Coma - - - - - - - - - - - - - - - - - - -
Death - - - - - - - - - - - - - - - - - - -

Body weight:

Body Weight
The body weights of all animals were increasing during the study. Stagnation of the body weight in 2 animals and mild increase of the body weight in the rest animals was observed between first and second week after administration. Summary results of body weight development are presented in Table 3.
Table 3 Body Weight
Sex Dose ID Body Weight (g) Body Weight Difference (g)
Initial Week 1 Week 2 Week 1 - Initial Week 2 - Initial Week 2 - Week 1
♀ 2000 mg/kg 1 180 190 195 10 15 5
2 170 185 190 15 20 5
3 175 190 200 15 25 10
4 160 180 180 20 20 0
5 180 190 190 10 10 0
6 165 180 190 15 25 10

Gross pathology:

8.4 Necropsy
All animals were necropsied. During necropsy, no macroscopic findings were noticed.

Table 4 Necropsy Results
Sex Dose ID Result Sex Dose ID Result
♀ 2000 mg/kg 1 no finding ♀ 2000 mg/kg 4 no finding
2 no finding 5 no finding
3 no finding 6 no finding

Other findings:

The test item Copper dioleate administered to 6 females at limit dose of 2000 mg/kg body weight did not cause death. The body weights of all animals were increasing during the study. Stagnation of the body weight in 2 animals and mild increase of the body weight in the rest animals was observed between first and second week after administration.
No signs of toxicity were observed at the dosage of 2000 mg/kg body weight during the first 4 hours in females or 14-day observation period. During necropsy, no macroscopic findings were noticed.
9 CONCLUSION
The LD50 of the test item Copper dioleate is higher than 2000 mg/kg body weight after single oral administration to Wistar rats.
Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423 it can be concluded that the test item Copper dioleate is classified in Category 5/Unclassified with a LD50 cut off value equal to or greater than 5000 mg/kg body weight, after single oral administration to Wistar rats.

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

it has a low acute oral toxicity (LD50 > 2000 mg/kg bw).

Executive summary:

The purpose of the study was to evaluate the potential toxic effect of the test item Copper dioleate

when administered as a single oral dose to Wistar rats.

The procedure according to OECD Guideline 423 Acute Toxic Class (ATC) method was used.

Available information indicated that the test item is likely to be non-toxic; therefore, a limit dose of

2000 mg/kg body weight was used as a starting dose. Two groups of 3 females were dosed. All 6

females survived the limit dose. The limit dose of 2000 mg/kg body weight did not cause death, or

evident signs of toxicity. Stagnation of the body weight in 2 animals and mild increase of the body

weight in the rest animals was observed between first and second week after administration.

The LD50 of the test item Copper dioleate is higher than 2000 mg/kg body weight after single oral

administration to Wistar rats.

Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD

Guideline 423 it can be concluded that the test item Copper dioleate is classified in Category

5/Unclassified with a LD50 cut off value equal to or greater than 5000 mg/kg body weight, after

single oral administration to Wistar rats.