3-[[3-[[(2-cyanoethyl)amino]methyl]-3,5,5-trimethylcyclohexyl]amino]propiononitrile

Acute Toxicity: dermal

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

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-[[3-[[(2-cyanoethyl)amino]methyl]-3,5,5-trimethylcyclohexyl]amino]propiononitrile

EC Number:

300-496-1

EC Name:

3-[[3-[[(2-cyanoethyl)amino]methyl]-3,5,5-trimethylcyclohexyl]amino]propiononitrile

Cas Number:

93940-97-7

Molecular formula:

C16H28N4

IUPAC Name:

3-[(3-{[(2-cyanoethyl)amino]methyl}-3,5,5-trimethylcyclohexyl)amino]propanenitrile

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 or test system 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:

other: 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 ani

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.