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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010-10-06 to 2011-01-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted in accordance with GLP regulations and OECD/EU guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
31 May 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
24 February 1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
August 1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
(2-methylpropylidene)[(3-{[(2-methylpropylidene)amino]methyl}cyclohexyl)methyl]amine
EC Number:
619-764-7
Cas Number:
173904-11-5
Molecular formula:
C16H30N2
IUPAC Name:
(2-methylpropylidene)[(3-{[(2-methylpropylidene)amino]methyl}cyclohexyl)methyl]amine
Test material form:
other: liquid

Test animals

Species:
rat
Strain:
other: Crl(WI)Br
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90.
- Hygienic level: SPF at arrival and kept in a good conventional environment during the study
- Justification of strain: The Wistar rats as a rodent is one of the standard species of acute toxicity studies.
- Number of animals: 5 animals/sex
- Age of animals: Young adult rats, 8 weeks old
- Body weight range (male): 282-292 g
- Body weight range (female): 261-276 g
- Acclimatisation time: 5 days
Animal Husbandry:
- Animal health: Only healthy animals were used for the study. The health status was certified by the breeder.
- Number of animal room: 5
- Housing: during acclimatization: 3 animals/sex/cage During the study: animals were housed individually.
- Cage type: Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Illumination: Artificial light, from 6 a.m. to 6 p.m.
- Temperature: 22 ± 3 °C
- Relative humidity: 30 - 70 %
- Ventilation: 8-12 air exchanges/hour by central air-condition system.
- Environmental conditions were maintained by an air-conditioning system.
- Temperature and relative humidity were verified and recorded daily during the study. Before housing the animals the microbiological status of the room was checked.
- Food and water supply: The animals received ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum. Animals received tap water from watering bottles ad libitum.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
The back of animals was shaven (approximately 10 % area of the total body surface) 24 hours prior to the treatment. The test item was applied in a single dose uniformly at least 10 % area of the total body surface throughout a 24-hour exposure period. Sterile gauze pads were placed on the skin of rats. These gauzes were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was wrapped with semi-occlusive plastic wrap for 24 hours. At the end of the exposure period, residual test item was removed, using body temperature water.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males/5 females per dose
Control animals:
no
Details on study design:
Dose Level: The test item was not expected to be lethal at 2000 mg/kg bw. A limit test was performed.
A single administration was performed by dermal route and was followed by a 14-day observation period.

Observations
Mortality
Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day.
Clinical Observations
Animals were observed individually 1 h and 5 h after dosing, and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body weight
The body weight were recorded on day 0 (shortly before the treatment), at day 7 and at day 15 on all animals with a precision of 1 g.
Pathology
All animals were subjected to gross pathology. All animals were exsanguinated under isoflurane anaesthesia. After examination of the external appearance the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross pathological changes were recorded for each animal on the post mortem record sheets.
Statistics:
NA

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day.
No mortality occurred after the 24-hour dermal exposure to Incorez 397 in Crl:(WI)BR male and female rats during the study.
Clinical signs:
other: Animals were observed individually 1 h and 5 h after dosing, and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomo
Gross pathology:
No macroscopic alterations of organs and tissues referred to the systemic toxic effect of the test item were seen during the necropsy. The external alterations (dry skin surface, crusting) were in line the observed local irritant symptoms.
Other findings:
It is to be noted that the test item caused dermal irritation response on the site of administration.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions, the acute dermal LD50 value of the test item Incorez 397 proved to be greater than 2000 mg/kg bw in male and female Crl:(WI)BR rats. On the other hand, it is to be noted that the test item caused dermal irritation response on the site of administration.
Executive summary:

An acute dermal toxicity study was performed with test item Incorez 397 in Crl:(WI)BR rats, in compliance with OECD Guideline No.: 402, Directive 92/69 EEC B.3 and OPPTS 870.1200.

A limit test was carried out. A single group of male and female animals (n=5 animals/sex) was exposed to Incorez 397 at 2000 mg/kg bw by dermal route. The test item was applied in original form and left in contact with the skin for 24 hours, followed by a 14-day observation period.

No mortality occurred during the study. Neither male nor female animals treated at 2000 mg/kg bw showed behavioural changes and no systemic toxic signs were noted during the study. The test item caused dermal irritation symptoms as slight to severe erythema, very slight oedema and other signs (dry skin surface, wounds, crusting) during 14-day observation period.

Slight body weight loss was observed in one female on first week. It could not be evaluated as a toxic effect of test item.

No macroscopic alterations of organs and tissues referred to the systemic toxic effect of the test item were seen during the necropsy. The external alterations (dry skin surface, crusting) were in line the observed local irritant symptoms.

Under the experimental conditions of this test, the acute dermal LD50 value of the test item Incorez 397 proved to be greater than 2000 mg/kg bw in male and female Crl:(WI)BR rats.