1,3-Cyclohexanedimethanamine, N1,N3-bis(2-methylpropylidene)-

Endpoint summary

Description of key information

Incorez 397 was tested for acute oral toxicity according to OECD Guideline 423/EU Method B.1 tris in a single oral dose of 2000 mg/kg bw in rats. No lethality was noted at single oral dose of 2000 mg/kg bw.
Incorez 397 was tested for acute dermal toxicity in a limit test in rats according to OECD Guideline 402/EU Method B.3. 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.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-11-05 to 2010-12-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

31 May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

17 December 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: Crl(WI)Br

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90.
- Age at study initiation: Young adult rats, 8 weeks old;
- Weight at study initiation: 218-232 g (first step); 220-229 g (second step);
- Fasting period before study: 24 hours;
- Housing: Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets; 3 animals/sex/cage;
- Diet: ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum;
- Water: tap water from watering bottles ad libitum;
- Acclimation period: 5 days in first step and 6 days in second step;

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C;
- Humidity: 30 - 70 %;
- Air changes: 8-12 air exchanges/hour by central air-condition system;
- Photoperiod: Artificial light, from 6 a.m. to 6 p.m.

Route of administration:

oral: gavage

Vehicle:

other: Helianthi Annui Oleum Raffinatum

Details on oral exposure:

A single oral administration - followed by a fourteen-day observation period - was performed by gavage. The day before treatment the animals were
fasted. The food but not water was withheld overnight. Animals were weighed before the application and the food was given back 3 hours after the
treatment.

Doses:

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 was met.

No. of animals per sex per dose:

3 + 3 females per dose (2000 mg/kg bw)

Control animals:

no

Details on study design:

Testing Procedure
A single oral administration - followed by a fourteen-day observation period - was performed by gavage. The day before treatment the animals were
fasted. The food but not water was withheld overnight. Animals were weighed before the application and the food was given back 3 hours after the treatment.

Dose Level
Starting dose was selected on the basis of the available information about the test item.
The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 was met.
A single administration will be performed by oral route and will be 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
Individual observations were performed on the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, 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.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death occurred at 2000 mg/kg single oral dose of Incorez 397. All female rats in step 1 and step 2 survived until the end of the 14-day observation period.

Clinical signs:

other: In group 1 treated with 2000 mg/kg dose clinical sign of reaction comprised of diarrhoea (2 cases out of 57 observations) and diuresis (6/57). Diarrhoea (score +1) occurred in one animal. Diuresis (score +2; +3) was found in one animal. One Animal was fr

Gross pathology:

All animals survived until the scheduled necropsy on Day 15.
Slight hydrometra was found in one animal of the group 1 (2000 mg/kg) and moderate hydrometra was observed in two females of the group 2 (2000 mg/kg) is physiological finding and connected to the cycle of the animal. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.

Other findings:

NA

Interpretation of results:

GHS criteria not met

Conclusions:

No lethality was noted at single oral dose of 2000 mg/kg bw.

Executive summary:

The acute toxic class method (OECD Guideline No. 423) was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 (presented in Annex VII) was met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on the 15th day after the treatment.

Lethality, Clinical symptoms and Body weight:

No lethality was noted at single oral dose of 2000 mg/kg bw. In first step, disturbances of the autonomic functions as diarrhoea and diuresis were observed between treatment day and Day 1. In second step, no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal. The body weight development was undisturbed in all animals.

Gross pathology:

All animals survived until the scheduled autopsy on Day 15. All organs of all experimental animals proved to be free of treatment related gross pathological changes.

Evaluation:

The method used is not intended to allow the calculation of a precise LD50 value. The test item is ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as below:

Dose (mg/kg bw)	Mortality (dead/treated)	LD50  (mg/kg bw)	GHS category
2000	0/6	above 2000	5

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The study was conducted in accordance with GLP regulations and OECD/EU guidelines.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

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.

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

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.

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

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.

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The study was conducted in accordance with GLP regulations and OECD/EU guidelines.

Additional information

Acute oral toxicity:

The acute toxic class method (OECD Guideline No. 423) was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 (presented in Annex VII) was met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on the 15th day after the treatment.

No lethality was noted at single oral dose of 2000 mg/kg bw. In first step, disturbances of the autonomic functions as diarrhoea and diuresis were observed between treatment day and Day 1. In second step, no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal. The body weight development was undisturbed in all animals. All animals survived until the scheduled autopsy on Day 15. All organs of all experimental animals proved to be free of treatment related gross pathological changes.

Evaluation:

The method used is not intended to allow the calculation of a precise LD50 value. Nevertheless, based on the available data, the LD50 could be determined to be greater than 2000 mg/kg bw.

 

Acute dermal toxicity:

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.

 

Acute inhalation toxicity:

The test for acute inhalation toxicity was waived. Testing for acute toxicity via the inhalation route was not applicable as data on acute oral and acute dermal toxicity were available. According to REACH Regulation No. 1907/2006, Annex VIII, 8.5 data for a maximum of two routes of exposure need to be provided.

Justification for classification or non-classification

No classification and labelling is required according to Regulation (EC) No 1272/2008 (CLP).