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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
3 October 2012 - 31 October 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
(see below)
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
(see below)
Principles of method if other than guideline:
On Day 6 of the study, one female was discovered to be pregnant and was therefore humanely killed. The data obtained was considered invalid and was not included for reporting purposes as the animal was not nulliparous and non-pregnant as stated in the guidelines. As no signs of toxicity were observed in the remaining animals, it was considered that treatment of an additional animal could not be justified as no further information was likely to be obtained. This deviation was considered not to affect the integrity or outcome of the study.
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
Tris[oxalate(2-)]dicerium
EC Number:
205-362-5
EC Name:
Tris[oxalate(2-)]dicerium
Cas Number:
139-42-4
Molecular formula:
C6Ce2O12
IUPAC Name:
4,4'',5,5''-tetraoxo-1',4'-dihydro-1λ³,1''λ³,3λ³,3''λ³-dispiro[1λ³,3λ³-dioxa-2-ceracyclopentane-2,5'-1λ³-oxa-3λ³-oxa-2-ceracyclopenta[4,5-d]1λ³-oxa-3λ³-oxa-2-ceracyclopentane-2',2''-[1λ³,3λ³]dioxa-[2]ceracyclopentane]-2',2',2',2,2,2-hexakis(ylium)-1',4',1,1'',3,3''-hexaide
Test material form:
solid: crystalline
Details on test material:
Description: White crystalline solid
Storage Conditions: Room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain: RccHan:WIST
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: At the start of the study the animals weighed at least 200 g. The weight variation did not exceed ± 20 % of the mean weight for each sex
- Fasting period before study: No
- Housing: The animals were housed in suspended solid-floor polypropylene cages furnished with woodflakes, individually during the exposure period and in groups of up to four, by sex, for the remainder of the study.
- Diet (e.g. ad libitum): ad libitum access to rodent diet
- Water (e.g. ad libitum): Free access to mains drinking water
- Acclimation period: At least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 °C
- Humidity (%): 30 - 70 % (relative)
- Air changes (per hr): At least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): lighting was controlled by a time switch to give twelve hours continuous light (0600 to 1800) and twelve hours darkness

IN-LIFE DATES: From: 3 October 2012 To: 30 October 2012

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
other: DMSO and distilled water
Details on dermal exposure:
TEST SITE
- Area of exposure: The back and flanks clipped free of hair.
- % coverage: Approximately 10 % of the total body surface area.
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair was wiped with cotton wool moistened with dimethyl sulphoxide followed by distilled water.
- Time after start of exposure: After the 24 hour exposure period, following the removal of the semi-occlusive dressing.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bodyweight.

VEHICLE
- Amount(s) applied: The test material was moistened with dimethyl sulphoxide follwed by distilled water prior to application.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
- Treatment schedule: In the absence of data suggesting toxicity, one male and one female were initially treated at a dose level of 2000 mg/kg. These animals remained in individual housing following exposure. As no mortalities were noted, a further group of animals (four males and four females) was similarly treated with the test material at a dose level of 2000 mg/kg bodyweight to give a total of five males and five females.
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for deaths or overt signs of toxicity 0.5, 1, 2 and 4 hours after dosing and subsequently once daily. Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: Yes. At the end of the study the animals were killed by cervical dislocation and subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
- Other examinations performed: After removal of the dressings and subsequently once daily, the test sites were examined for evidence of primary irritation and scored according to the Draize scale:

Erythema and Eschar Formation Value
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to slight eschar formation (injuries in depth) 4

Oedema Formation
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4

Any other skin reactions, if present, were also recorded.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There was no mortality.
Clinical signs:
other: No signs of systemic toxicity were observed.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
DERMAL REACTIONS
Very slight erythema was noted at the test site of three animals. One male and one female exhibited erythema scores of 1 on Days 1 to 3. A further female exhibited an erythema score of 1 on Day 1. No other signs of dermal irritation were observed.

FURTHER INFORMATION
On Day 6 of the study, one female was discovered to be pregnant and was therefore humanely killed. The data obtained was considered invalid and was not included for reporting purposes as the animal was not nulliparous and non-pregnant as stated in the guidelines. As no signs of toxicity were observed in the remaining animals, it was considered that treatment of an additional animal could not be justified as no further information was likely to be obtained. This deviation was considered not to affect the integrity or outcome of the study.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 of the test material in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight and, as such, the test material is not classified in accordance with EU criteria.
Executive summary:

The acute dermal toxicity of the test material was assessed in a limit test conducted in the Wistar strain rat in accordance with the standardised guidelines OECD 402 and EU Method B.3.

Initially, two animals (one male and one female) were given a single, 24 hour, semi-occluded dermal application of the test material to intact skin at a dose level of 2000 mg/kg bodyweight. Based on the results of the initial test, a further group of eight animals (four males and four females) was similarly treated. Clinical signs and bodyweight development were monitored during the 14 day observation period. All animals were subjected to gross necropsy.

There were no deaths and no signs of systemic toxicity. Furthermore, the animals showed the expected gains in bodyweight. Very slight erythema was noted at the test sites of three animals, though no other signs of dermal irritation were noted. No abnormalities were noted at necropsy.

Therefore, under the conditions of the study, the LD50 of the test material in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.