3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol

Endpoint summary

Description of key information

Acute oral toxicity: 

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

Acute Inhalation toxicity:

For test chemical, acute toxicity testing by the inhalation route was considered for waiver given that the substance has low vapour pressure of 0.00162 Pa at 25 degree C . Thus, exposure by inhalation is also unlikely for test chemical given the comparitively larger size of the particulates.

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) for test chemical was considered based on experimental study, the value considered to be >2000 mg/kg bw in rabbits. Thus, comparing this value with the criteria of CLP regulation, 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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from study report

Qualifier:

according to guideline

Guideline:

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

Principles of method if other than guideline:

The aim of this study was to assess the toxicity potential of test chemical after single oral administration in rats.

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS

- Source:In-House Bred
- Age at study initiation:8- 11 weeks at the time of dosing
- Weight at study initiation: 168-192 g (Individual body weights were within ± 5% prior to treatment after overnight fasting)
- Fasting period:The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing.
- Housing: The animals were housed individually in polycarbonate cages with corn cobs as bedding in a controlled environment.
- Diet (e.g. ad libitum): Conventional laboratory rodent diet (Nutrivet Life Sciences, Pune), ad libitum
- Water (e.g. ad libitum): Aqua guard filtered tap water, ad libitum
- Acclimation period: Animal numbers 1-3 were acclimatized for five days, and 4-6 for seven days prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):20.40 - 23.10°C
.- Humidity (%):Minimum: 37.40 - 56.00%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12-hrs light/12-hrs dark

Route of administration:

oral: unspecified

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage):10 ml
- Justification for choice of vehicle: Corn oil was selected because test item was not miscible in distilled water.
- Lot/batch no. (if required): MKBD4650- Purity: No data av available

MAXIMUM DOSE VOLUME APPLIED:10 ml/kg body weight

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 female rats

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical Observation, body weight and mortality.

Statistics:

No data available

Preliminary study:

No data available

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other:

Remarks:

No mortality observed

Mortality:

No mortality was observed in the animals treated with 2000 mg/kg throught out the 14 days observation period post dosing.

Clinical signs:

other: At 2000 mg/kg, animal nos. 1, 2, 3, 4 and 6 were observed normal at 30 minutes, mild to moderate diarrhea at 1, 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. Additionally, animal no. 2 was observed with

Gross pathology:

No external or internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice.

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.	Group/ Dose (mg/kg)	Body Weight (gram)			Body Weight Change (%)
		Day 0	Day 7	Day 14	Day 0-7	Day 0-14
1	G1/ 2000	174	193	216	10.92	24.14
2		171	185	210	8.19	22.81
3		168	192	202	14.29	20.24
4		192	205	218	6.77	13.54
5		178	149	178	-16.29	0.00
6		174	189	198	8.62	13.79

 

 

Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)

Sex:Female

Group/ Dose (mg/kg)		Rats Body Weight (g)			Body Weight Changes (%)
		Day 0	Day 7	Day 14	0-7	0-14
G1/ 2000	Mean	176.17	185.50	203.67	5.42	15.75
	SD	8.45	19.10	14.77	10.95	8.91
	n	6	6	6	6	6

Keys:SD = Standard Deviation, n = Number of Animals

Table 3: Individual Animal Clinical Signs and Symptoms

Sex:Female

Animal No.	Group/ Dose (mg/kg)	Hours (Day 0)
		1/2	1	2	3	4
1	G1/ 2000	1	49^++	49^+	64^+ 49^+	64^+ 49^+
2		1	49^++	49^+	64^+ 49+	64^+ 49^+
3		1	49^++	49^+	64^+ 49^+	64^+ 49^+
4		1	49^+	49^++	64^+ 49^++	64^+ 49^+
5		1	1	49^+	64^+ 49^+	49^+ 64^+
6		1	49^++	49^++	64^+ 49^+	49^+ 64^+

Animal No.	Group/ Dose (mg/kg)	Days post dosing
		1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	G1/ 2000	1	1	1	1	1	1	1	1	1	1	1	1	1	1
2		182	1	1	1	1	1	1	1	1	1	1	1	1	1
3		1	1	1	1	1	1	1	1	1	1	1	1	1	1
4		1	1	1	1	1	1	1	1	1	1	1	1	1	1
5		1	1	1	1	1	1	1	1	1	1	1	1	1	1
6		1	1	1	1	1	1	1	1	1	1	1	1	1	1

Keys:1 = Normal, 49 = Diarrhoea, 64 = Epistaxis, 182 = Soiled anal region with fecal material,⁺= Mild,⁺⁺= Moderate

 

Table 4: Individual Animal Mortality Record

Sex:Female

Animal No.	Group/ Dose (mg/kg)	Day of Observation (Day 0 to 14)
		Morning Observations	Evening Observations
1	G1/ 2000	No mortality and morbidity	No mortality and morbidity
2		No mortality and morbidity	No mortality and morbidity
3		No mortality and morbidity	No mortality and morbidity
4		No mortality and morbidity	No mortality and morbidity
5		No mortality and morbidity	No mortality and morbidity
6		No mortality and morbidity	No mortality and morbidity

  

Table 5: Gross Necropsy Observation

 Sex:Female  

Animal No.	Group/ Dose (mg/kg)	Mode of Death	Gross Observation
			External	Internal
1	G1/ 2000	Terminal sacrifice	No abnormality detected	No abnormality detected
2		Terminal sacrifice	No abnormality detected	No abnormality detected
3		Terminal sacrifice	No abnormality detected	No abnormality detected
4		Terminal sacrifice	No abnormality detected	No abnormality detected
5		Terminal sacrifice	No abnormality detected	No abnormality detected
6		Terminal sacrifice	No abnormality detected	No abnormality detected

 

 

Interpretation of results:

other: Not classified

Conclusions:

Body weight gain was observed in all the animals treated with 2000 mg/kg body weight, during the 14-day observation period. At 2000 mg/kg, animal number 1, 2, 3, 4 and 6 were observed normal at 30 minutes, mild to moderate diarrhea at 1, 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. Additionally, animal number 2 was observed with soiled anal region with fecal material on day1. Animal number 5 was observed normal at 30 minutes and 1 hour, mild diarrhea at 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice
The acute oral LD50 value for the test chemical was considered to be greater than 2000 mg/kgbw.

Executive summary:

An acute oral toxicity study of test chemical was conducted in female Wistar rats as per OECD No. 423.Six Wistar female rats were for the acute oral toxicity study. Prior to dosing, the animals were fasted for minimum 16-18 hours and for 3-4 hours post dosing, with food withheld but drinking water provided ad libitum. Corn oil was selected as the vehicle because test item was not miscible in distilled water. Three rats of the first group were dosed with starting dose of 2000mg/kgbw and the animals didnot show any mortality so another three rats of the same group were dosed with 2000 mg/kgbw and no mortality was observed. Hence, further dosing was stopped. Body weight gain was observed in all the animals treated with 2000 mg/kg body weight, during the 14-day observation period. At 2000 mg/kg, animal number 1, 2, 3, 4 and 6 were observed normal at 30 minutes, mild to moderate diarrhea at 1, 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. Additionally, animal number 2 was observed with soiled anal region with fecal material on day1. Animal number 5 was observed normal at 30 minutes and 1 hour, mild diarrhea at 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice. The acute oral LD50 value for the test chemical was considered to be greater than 2000 mg/kgbw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is klimish 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:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Clinical signs:

other: 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 study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

The objective of the study was to assess the dermal toxicity of test chemical after single dose application by dermal route in rats.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: In-House Bred
- Age at study initiation: No data available-
- Weight (Prior to Treatment):Male: 225-245 g ; Female: 215-231 g
- Fasting period before study: No data available
- Housing:The animals were housed individually in polycarbonate cages with corn cob as bedding in a controlled environment
.- Diet (e.g. ad libitum): Conventional laboratory rodent diet (Nutrivet Life Sciences, Pune),ad libitum
- Water (e.g. ad libitum): Aqua guard filtered tap water, ad libitum
- Acclimation period: All animals were acclimatized to the test conditions for 5 days prior to administration of the test item.ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.20-24.40°C
- Humidity (%): 36.60-57.30%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12-hrs light/:12-hrs dark

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE

- Area of exposure: The test item was applied uniformly over clipped dorsal area of rat skin.
- % coverage: Approximately 10% body surface area of rat.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The residual test item was removed by using distilled water.
- Time after start of exposure: 24-hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): A limit dose of 2000 mg/ kg body weight of test item was applied.
- Constant volume or concentration used: Yes
- For solids, paste formed: No VEHICLE
- Amount(s) applied (volume or weight with unit): No data available
- Concentration (if solution): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

Duration of exposure:

24 hours

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

Total: 10 rats 2000 mg/kg: 5 males, 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily
- Necropsy of survivors performed: Yes
.- Other examinations performed: Clinical signs, body weight, local signs/skin reactions and mortality.

Statistics:

No statistical analysis was performed since the study was terminated with limit test.

Preliminary study:

No data available

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Non toxic to animals

Mortality:

No mortality was observed at 2000 mg/kg body weight during the 14 day observation period.

Clinical signs:

other: No systemic or local signs of toxicity were observed at 2000 mg/kg body weight during the experimental period.

Gross pathology:

The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Other findings:

No data available

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

 

Dose:2000 mg/ kg bodyweight                                                                                                                Density:0.96365

Animal No.	Sex	Dose (ml) Applied*	Body Weight (gram)			Body Weight Change (%)
			Day 0	Day 7	Day 14	Day 0-7	Day 0-14
1	Male	0.49	236	243	249	2.97	5.51
2		0.48	231	239	247	3.46	6.93
3		0.47	225	248	274	10.22	21.78
4		0.51	245	270	287	10.20	17.14
5		0.48	233	240	260	3.00	11.59
6	Female	0.45	215	223	215	3.72	0.00
7		0.47	227	231	233	1.76	2.64
8		0.48	231	228	233	-1.30	0.87
9		0.48	231	236	237	2.16	2.60
10		0.45	215	213	217	-0.93	0.93

Key:* = Based on density of test item and day 0 body weight taken prior to dose application.

 

Table 2: Individual Animal Clinical Signs and Symptoms

 

Dose:2000 mg/kg body weight

Animal No.	Sex	Hour(s) - Day 0				Day
		1	2	3	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	Male	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
2		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
3		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
4		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
5		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
6	Female	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
7		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
8		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
9		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
10		1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1

Key: 1 = Normal

 

Table 3: Individual Animal Mortality Record

 

Dose:2000 mg/kg body weight

Animal No.	Sex	Days of Observation (0 to 14)
		Morning Observations	Evening Observations
1	Male	No mortality and morbidity	No mortality and morbidity
2		No mortality and morbidity	No mortality and morbidity
3		No mortality and morbidity	No mortality and morbidity
4		No mortality and morbidity	No mortality and morbidity
5		No mortality and morbidity	No mortality and morbidity
6	Female	No mortality and morbidity	No mortality and morbidity
7		No mortality and morbidity	No mortality and morbidity
8		No mortality and morbidity	No mortality and morbidity
9		No mortality and morbidity	No mortality and morbidity
10		No mortality and morbidity	No mortality and morbidity

 

Table 4:Summaryof Animal Body Weight (g) and Body Weight Changes (%)

 

Dose:2000 mg/kg body weight

Sex		Body Weight (gram)			Body Weight Changes (%)
		Day 0	Day 7	Day 14	Day 0-7	Day 0-14
Male	Mean	234.00	248.00	263.40	5.97	12.59
	SD	7.35	12.79	17.01	3.88	6.86
	n	5	5	5	5	5
Female	Mean	223.80	226.20	227.00	1.08	1.41
	SD	8.20	8.76	10.20	2.14	1.17
	n	5	5	5	5	5

Keys:SD= Standard deviation, n = Number of animals

 

Table 5: GrossNecropsyObservation

 

 Dose:2000 mg/kg body weight                                                                             Mode of Death:Terminal Sacrifice

Animal No.	Sex	Gross Observation
		External	Internal
1	Male	No abnormalities detected	No abnormalities detected
2		No abnormalities detected	No abnormalities detected
3		No abnormalities detected	No abnormalities detected
4		No abnormalities detected	No abnormalities detected
5		No abnormalities detected	No abnormalities detected
6	Female	No abnormalities detected	No abnormalities detected
7		No abnormalities detected	No abnormalities detected
8		No abnormalities detected	No abnormalities detected
9		No abnormalities detected	No abnormalities detected
10		No abnormalities detected	No abnormalities detected

 

Interpretation of results:

other: Not classified

Conclusions:

No mortality was observed at 2000 mg/kg body weight during the 14 day observation period. No systemic or local signs of toxicity were observed at 2000 mg/kg body weight during the experimental period. The male and female animals were observed with body weight gain compared to day 0 throughout the experiment. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Based on the results, the acute dermal median lethal dose of test chemical was considered to be greater than 2000 mg/kg body weight.

Executive summary:

The acute toxicity of test chemical was determined after single dose application by dermal route in rats. This study was performed according to OECD 402 Guidelines. Five male and five female Wistar rats were treated with test item by a single dermal application at the dose level of 2000 mg/kg body weight and observed for 14 days after treatment. Since no test item related mortality was observed, the study was terminated with limit test only. The test item was applied uniformly over clipped dorsal area of rat skin. The porous gauze dressing was put on the intact skin and wrapped around the abdomen and it was anchored with non-irritating tape. After 24 hour application period, the residual test item was removed by using distilled water. The skin reactions were assessed. The animals were observed daily for clinical signs, mortality. The body weight changes were recorded on day 0 and on day 7 and day 14. All animals were necprosied and examined macroscopically. No statistical analysis was performed since the study was terminated with limit test. No mortality was observed at 2000 mg/kg body weight during the 14 day observation period. No systemic or local signs of toxicity were observed at 2000 mg/kg body weight during the experimental period. The male and female animals were observed with body weight gain compared to day 0 throughout the experiment. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Based on the results, the acute dermal median lethal dose of test chemical was considered to be greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is klimish 1 and from study report

Additional information

Acute oral toxicity:

An acute oral toxicity study of test chemical was conducted in female Wistar rats as per OECD No. 423. Six Wistar female rats were for the acute oral toxicity study. Prior to dosing, the animals were fasted for minimum 16-18 hours and for 3-4 hours post dosing, with food withheld but drinking water provided ad libitum. Corn oil was selected as the vehicle because test item was not miscible in distilled water. Three rats of the first group were dosed with starting dose of 2000mg/kgbw and the animals didnot show any mortality so another three rats of the same group were dosed with 2000 mg/kgbw and no mortality was observed. Hence, further dosing was stopped. Body weight gain was observed in all the animals treated with 2000 mg/kg body weight, during the 14-day observation period. At 2000 mg/kg, animal number 1, 2, 3, 4 and 6 were observed normal at 30 minutes, mild to moderate diarrhea at 1, 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. Additionally, animal number 2 was observed with soiled anal region with fecal material on day1. Animal number 5 was observed normal at 30 minutes and 1 hour, mild diarrhea at 2, 3 and 4 hours and mild epistaxis at 3 and 4 hours followed by normal clinical sign till day 14. No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice The acute oral LD50 value for the test chemical was considered to be greater than 2000 mg/kgbw.

Acute Inhalation toxicity:

For test chemical, acute toxicity testing by the inhalation route was considered for waiver given that the substance has low vapour pressure of 0.00162 Pa

at 25 degrees C. Thus, exposure by inhalation is also unlikely for test chemical given the comparitively larger size of the particulates.

Acute Dermal Toxicity:

The acute toxicity of test chemical was determined after single dose application by dermal route in rats. This study was performed according to OECD 402 Guidelines. Five male and five female Wistar rats were treated with test item by a single dermal application at the dose level of 2000 mg/kg body weight and observed for 14 days after treatment. Since no test item related mortality was observed, the study was terminated with limit test only. The test item was applied uniformly over clipped dorsal area of rat skin. The porous gauze dressing was put on the intact skin and wrapped around the abdomen and it was anchored with non-irritating tape. After 24 hour application period, the residual test item was removed by using distilled water. The skin reactions were assessed. The animals were observed daily for clinical signs, mortality. The body weight changes were recorded on day 0 and on day 7 and day 14. All animals were necropsied and examined macroscopically. No statistical analysis was performed since the study was terminated with limit test. No mortality was observed at 2000 mg/kg body weight during the 14 day observation period. No systemic or local signs of toxicity were observed at 2000 mg/kg body weight during the experimental period. The male and female animals were observed with body weight gain compared to day 0 throughout the experiment. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Based on the results, the acute dermal median lethal dose of test chemical was considered to be greater than 2000 mg/kg body weight.

 

Justification for classification or non-classification

Based on the above experimental studies on test chemical indicate that the test chemical is not likely to cause any toxicity when exposed to test organisms via oral, dermal or inhalation route of exposure. Hence, the test chemical can be classified under the category "Not Classified" as per CLP Regulation.