Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
03/2018 - 06/2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Specific details on test material used for the study:
Test item No.: 13/0241-4
Batch identification: 16315004
CAS No.: 93940-97-7
Purity: 96.1 area-% (mixture of isomers)
Homogeneity: The test item was homogeneous by visual inspection.
Storage stability: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
Expiry date: May 01, 2019
Storage conditions: Ambient (RT)
Physical state/ color: Liquid / yellowish, clear
Density [g/mL]: 0.986 (determined by Bioassay Laboratories)

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Age on day 0: Young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks)
Body weight on day 0: Animals of comparable weight (± 20% of the mean weight)

Room temperature / relative humidity: The animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 22°C  3°C for temperature and of 30 – 70% for relative humidity. There were no deviations from these ranges, which influenced the results of the study.
Air changes per hour: Approx. 10
Day / night rhythm: 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)
Type of cage: Makrolon cage, type III
Number of animals per cage: Single housing
Feeding: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
Drinking water: Tap water ad libitum
Bedding: H 15005-29; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)
Enrichment: Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Route of application: Single application to the clipped skin (dorsal and dorsolateral parts of the trunk); covering of the application site with a semi-occlusive dressing* for 24 hours. Afterwards removal of the semi-occlusive dressing, rinsing of the application site with warm water
Application area: About 40 cm² (corresponds to at least 10% of the body surface)
Duration of exposure:
24 hours
Doses:
limit dose: 2000 mg/kg bw
No. of animals per sex per dose:
5
Details on study design:
Observation period: 14 days

Body weight determination: Individual body weights shortly before application (day 0), weekly thereafter and on the last day of observation.

Clinical observations: Clinical signs for each animal were recorded several times on the day of application and at least once during each workday thereafter.

Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter /several times until the last day of observation / and on the last day of observation.

Mortality: A check for any dead or moribund animals was made at least once each workday, these records are archived by Bioassay.

Pathology: Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Systemic effects:
No systemic clinical signs were observed during clinical examination in male and female.

Local effects:
Male: moderate erythema (grade 2) was seen in four male animals from study day 1 until study day 3 and persisted in one of these animals until study day 10. Furthermore, incrustations were observed in four male animals from study day 2 until study day 3 and persisted in one of these animals until study day 10.

Female: Moderate erythema (grade 2) was seen in four female animals from study day 1 until study day 3 and persisted in one of these animals until study day 10. Furthermore, incrustations were observed in three female animals from study day 2 until study day 3, while in one of these animals scaling was noticed from study day 6 until study day 10.
Body weight:
All animals gained weight in a normal range throughout the study period.
Gross pathology:
No macroscopic pathologic abnormalities were noted in any animal examined on the last day of observation (5 males and 5 females).

Applicant's summary and conclusion

Conclusions:
In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the undiluted test item 3-[[3-[[(2-cyanoethyl)amino]methyl]-3,5,5- trimethylcyclohexyl]amino]propiononitrile to the clipped application site (dorsal and dorso-lateral parts of the trunk and covered by semi-occlusive dressing during the 24-hour exposure period. The application area comprised at least 10% of the total body surface. The animals were observed for 14 days.
- No mortality occurred
- No signs of systemic toxicity were observed
- The following test item-related local skin effects were recorded during the course of the study, local effects occurred within 10 days after application:
- Well-defined erythema (grade 2)
- Incrustations
- Scaling
- All animals gained weight in a normal range throughout the study period.
- No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study (5 males and 5 females).
Accordingly, the acute dermal median lethal dose (LD50) was determined to be LD50, dermal, rat > 2000 mg/kg bw
Executive summary:

Under the conditions of this study the median lethal dose (LD50) of 3-[[3-[[(2-cyanoethyl)amino]methyl]-3,5,5- trimethylcyclohexyl]amino]propiononitrile after dermal application was found to be greater than 2000 mg/kg bw in male and female rats.