[[4-[[2-(4-cyclohexylphenoxy)ethyl]ethylamino]-2-methylphenyl]methylene]malononitrile

Endpoint summary

Description of key information

Acute oral toxicity: 

Acute oral toxicity dose (LD50) was considered based on experimental study conducted on rats for the given test chemical. The LD50 value was considered is >5000 mg/kg bw. The study concluded that the LD50 value is >2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation toxicity: 

The acute toxicity inhalation study need not be conducted because exposure to humans via inhalation route is not likely taking into account the low vapour pressure of the substance, which is reported as 3.07E-010 Pa. Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore this study is considered for waiver.

 

Acute Dermal toxicity: 

The acute dermal toxicity dose (LD50) was considered based on experimental study conducted on rats for the test chemical. The LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

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:

August 1978

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Justification for type of information:

Data is from study report.

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

Acute toxicological investigations in female Wistar rats were conducted after oral administration of test chemical.

GLP compliance:

not specified

Test type:

other: not specified

Limit test:

no

Species:

rat

Strain:

Wistar

Remarks:

SPF-bred Wistar rats (strain Bor: WISW (SPF Cpb)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: about 14 weeks
- Weight at study initiation: initial weight was 150 to 200 g
- Identification: The anirnals were identified by marking their coat with picric acid.
- Housing: The rats were housed in groups of five animals each under conventional conditions in Makrolon Type-I11 cages on dust-free wood granules
- Diet (e.g. ad libitum): Feed (Altromin R 1324, producer: Altromin GmbH, Lage) was also available ad libitum.
- Water (e.g. ad libitum): tap water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1.5° C
- Humidity (%): about 60 ± 5 %.
- Photoperiod (hrs dark / hrs light): a 12-hour light/dark cycle (artificial light from 7 a.m. to 7 p.m. CET)

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 5000 mg/kg
- Amount of vehicle (if gavage): 30 ml/kg

DOSAGE PREPARATION (if unusual): The substance was formulated in tap water at room temperature.

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

10 female animals

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were inspected several times on the day of administration, and twice daily during the following 14-day observation period (once on weekends and bank holidays). During inspections, the type, onset, duration, and intensity of clinical signs were recorded.
- Body weight: The animals were weighed individually at application and in groups at the end of the 14-day observation period.
- Necropsy of survivors performed: yes, the animals were sacrificed.

Statistics:

not specified

Preliminary study:

not specified

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No animal died.

Clinical signs:

other: No female animal showed any signs of poisoning after Single administration of 5000 mg/kg.

Gross pathology:

not specified

Other findings:

not specified

Interpretation of results:

other: Not classified

Conclusions:

The acute oral toxicity dose (LD50) value was considered to be >5000 mg/kg bw, when 10 female SPF-bred Wistar rats were treated with the given test chemical via oral gavage route.

Executive summary:

The acute oral toxicity study was conducted by using given test chemical in 10 female SPF-bred Wistar rats at the dose concentration of 5000 mg/kg bw.

The acute toxicity experiment was carried out with SPF-bred Wistar rats (strain Bor: WISW (SPF Cpb), bred by Winkelmann, Borchen, Germany). At the start of study the female rats were about 14 weeks of age; their initial weight was 150 to 200 g.

The substance was formulated in tap water at room temperature and once administered to 10 female animals at a constant application volurne of 30 ml/kg. The animals were inspected several times on the day of administration, and twice daily during the following 14-day observation period (once on weekends and bank holidays). During inspections, the type, onset, duration, and intensity of clinical signs were recorded. The animals were weighed individually at application and in groups at the end of the 14-day observation period. Subsequently the animals were sacrificed.

No animal died at 5000 mg/kg bw. No female animal showed any signs of poisoning after Single administration of 5000 mg/kg. Body weight development was not affected.

Under the condition of this study, the acute oral toxicity dose (LD50) value was considered to be >5000 mg/kg bw, when 10 female SPF-bred Wistar rats were treated with the given test chemical via oral gavage route.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from study report.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Quality of whole database:

Waiver

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

This study was designed to determine the dermal LD50 of the test item (up to 2000 mg/kg) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: Young adult female rats aged between 8 - 10 weeks were used.
- Weight at study initiation: The weight range of approximately 216.5 to 228.3 grams at initiation of dosing.
Body weights at the start : Female Mean: 221.78 g (= 100 %); Minimum : 216.5 g (-2.38 %); Maximum : 228.3 g (+ 2.94 %)
- Identification: Each rat was individually identified by the cage number.
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.0 to 21.8 degree centigrade.
- Humidity (%): 53.7% to 59.4%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 01-03-2018 to 25-07-2018

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: the trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: approximately 10% of the total body surface area
- Type of wrap if used: The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.
- Time after start of exposure: 24 hours

Duration of exposure:

24 hours

Doses:

Dose Range Finding Study:
Group I : 200 mg/kg
Group I : 1000 mg/kg
Group I : 2000 mg/kg
Main Study: Group II : 2000 mg/kg

No. of animals per sex per dose:

Dose Range Finding Study:
Group I : 200 mg/kg - 1
Group I : 1000 mg/kg - 1
Group I : 2000 mg/kg - 1
Main Study: Group II : 2000 mg/kg - 2

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: yes, necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).

- Other examinations performed: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality, until sacrifice. Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.

- Evaluation of Dermal Reaction: Dermal reaction was observed daily for study period of 14 days.
- Body weights: Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.
- Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

Statistics:

not specified

Preliminary study:

Dose Finding Study: A single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered at the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Dose Range Finding Study: All animals survived through the study period of 14 days at 200 mg/kg, 1000 mg/kg and 2000 mg/kg body weight .
Main Study: Group II : All animals survived through the study period of 14 days.

Clinical signs:

other: Dose Range Finding Study: Animals treated at the dose level of 200 mg/kg, 1000 mg/kg and 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. Main Study: Group II : Animals treated at the dose level of 2000 m

Gross pathology:

Gross pathological examination did not reveal any abnormalities in animals from 200 mg/kg, 1000 mg/kg and 2000 mg/kg dose groups from dose range finding study and main study sacrificed terminally.

Other findings:

Dose Range Finding Study: Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

 

Laboratory Test Item Code :TAS/122/071

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	200	No clinical signs observed	1	1	Day 0 - Day 14	0/1

 

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	1000	No clinical signs observed	1	2	Day 0 - Day 14	0/1

 

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	1	3	Day 0 - Day 14	0/1

 

Main Study:

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
II	2000	No clinical signs observed	2	4, 5	Day 0 - Day 14	0/2

Table No. II 

Summary of Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/071

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
I	200	No dermal reaction observed	1	1	Day 0 - Day 14

 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
I	1000	No dermal reaction observed	1	2	Day 0 - Day 14

 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
I	2000	No dermal reaction observed	1	3	Day 0 - Day 14

 

Main Study:

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
II	2000	No dermal reaction observed	2	4, 5	Day 0 - Day 14

 

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/071

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	200	Mean	216.5	228.5	5.54	239.7	4.90	10.72
		± SD	-	-	-	-	-	-

 

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	1000	Mean	220.3	228.6	3.77	236.6	3.50	7.40
		± SD	-	-	-	-	-	-

 

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	221.6	233.7	5.46	240.3	2.28	8.44
		± SD	-	-	-	-	-	-

 

Main Study:

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	2000	Mean	225.25	235.30	4.47	243.75	3.59	8.23
		± SD	4.31	1.41	1.37	0.21	0.53	1.98

Table No.IV

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/071

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	200	1	TS	No abnormality detected

 

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	1000	2	TS	No abnormality detected

 

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	3	TS	No abnormality detected

 

                    Main Study:

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	2000	4, 5	TS	No abnormality detected

 

                     TS = Terminal Sacrifice

Interpretation of results:

other: Not classified

Conclusions:

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Executive summary:

The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals.

Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from study report.

Additional information

Acute oral toxicity:

The acute oral toxicity study was conducted by using given test chemical in 10 female SPF-bred Wistar rats at the dose concentration of 5000 mg/kg bw.

The acute toxicity experiment was carried out with SPF-bred Wistar rats (strain Bor: WISW (SPF Cpb), bred by Winkelmann, Borchen, Germany). At the start of study the female rats were about 14 weeks of age; their initial weight was 150 to 200 g.

The substance was formulated in tap water at room temperature and once administered to 10 female animals at a constant application volume of 30 ml/kg. The animals were inspected several times on the day of administration, and twice daily during the following 14-day observation period (once on weekends and bank holidays). During inspections, the type, onset, duration, and intensity of clinical signs were recorded. The animals were weighed individually at application and in groups at the end of the 14-day observation period. Subsequently the animals were sacrificed.

No animal died at 5000 mg/kg bw. No female animal showed any signs of poisoning after Single administration of 5000 mg/kg. Body weight development was not affected.

Under the condition of this study, the acute oral toxicity dose (LD50) value was considered to be >5000 mg/kg bw, when 10 female SPF-bred Wistar rats were treated with the given test chemical via oral gavage route.

Acute Inhalation toxicity: 

The acute toxicity inhalation study need not be conducted because exposure to humans via inhalation route is not likely taking into account the low vapour pressure of the substance, which is reported as 3.07E-010 Pa. Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore this study is considered for waiver.

 

Acute Dermal toxicity:

The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals.

Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral toxicity and acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral and acute dermal toxicity. For acute inhalation toxicity waiver was added so, not possible to classify.