Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: Key study. Test method according to OECD 423, GLP study. The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off may be considered to be 5000 mg/kg body weight by oral route in the rat.

Acute inhalation toxicity: Data waiving (study scientifically not necessary): According to REACH Annex VIII, column 2: In addition to the oral route, for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. The information is provided for dermal route.

Acute dermal toxicity: Key study. Test method according to OECD 402, GLP study. The LD50 of the test item was found to be higher than 2000 mg/kg body weight by dermal route in rats.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 November 2018 - 13 December 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-house bred animals
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 10 weeks
- Weight at study initiation: 177.28-205.76 (minimum and maximum mean weights of the 4 groups tested)
- Fasting period before study: Animals were fasted overnight (16 to 18 hours) prior to dosing. Water was provided ad libitum during fasting period. Feed was offered 3 to 4 hours after dosing.
- Housing: Three animals were housed in standard polypropylene cage (Size: L 430 × B 285 × H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized paddy husk was provided as bedding material.
- Diet (e.g. ad libitum): Ad libitum. Altromin Maintenance Diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG).
- Water (e.g. ad libitum): Ad libitum. Deep bore-well water passed through Reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes.
- Acclimation period: the animals were acclimatized for at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3ºC to 22.8ºC
- Humidity (%): 46 to 66%
- Air changes (per hr): 12-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL and 200 mg/mL
- Amount of vehicle (if gavage): 10 mL
- Justification for choice of vehicle: As per the in-house miscibility test, test item was miscible in corn oil. Corn oil is universally accepted and routinely used vehicle in oral toxicity studies.
- Lot/batch no. (if required): A1712001
- Purity: not reported

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The animals were dosed in a stepwise procedure with three female animals per step. A starting dose of 300 mg/kg body weight was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg body weight since the LD50 for test item was unavailable.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
6 (3 female rats per step. 2 steps per dose were conducted as per OECD TG 423)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At each step, the animals were observed for clinical signs of toxicity and mortality at 30 to 40 min, 1 hr (±10 mins), 2 hrs (±10 mins), 3 hrs (±10 mins) and 4 hrs (±10 mins) post dosing on day 1 and once daily thereafter for clinical signs of toxicity and twice daily for mortality during the 14 days observation period. The body weights were recorded on day 1 (before test item administration) and on day 8 and 15 during the observation period.
- Necropsy of survivors performed: yes. At termination, gross pathological findings were recorded and reported.
- Other examinations performed:
Clinical observations included: changes in skin, fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study.
Clinical signs:
No clinical signs of toxicity were observed in Step-I and Step-I confirmation animals dosed with 300 mg/kg bw.
In Step-II and Step-II confirmation, animals were dosed with 2000 mg/kg bw. No clinical signs were observed on day 1; diarrhea and wet perineum was observed on day 2 and wet perineum was observed on day 3. The observed clinical signs reversed back to normal by day 4 observation.
Body weight:
No changes were observed in body weight and percent change in body weight with respect to day 1. All the animals revealed physiologically normal increase in the body weight.
Gross pathology:
No gross pathological changes were observed in any of the animals.

Table 1: Individual animal clinical signs of toxicity and mortality record

Study Steps

&

Dose

(mg/kg body weight)

Animal No.

Sex

Clinical Signs of Toxicity and Mortality on

Day 1

Clinical Signs of Toxicity and Mortality on Day

30 to 40

min

1 hr

(±10 min)

2hrs

(±10 min)

3hrs

(±10

min)

4hrs

(±10

min)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Step-I

&

300

Rd2486

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2487

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2488

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Step-I

Confirmation &

300

Rd2489

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2490

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2491

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Step-II

&

2000

Rd2492

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2493

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2494

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Step-II

Confirmation &

2000

Rd2495

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2496

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2497

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

N: Normal; F: Female; min/mins: minutes; hr/hrs: Hour/Hours; 36: Diarrhea; 110: Wet perineum

Table 2: Individual animal body weight (g) and percent change in body weight with respect to day 1

Study Steps

&

Dose

(mg/kg body weight)

Animal No.

Sex

Body Weight (g) on Day

 

Percent Change in Body Weight with Respect to Day

1

8

15

 

1 to 8

1 to 15

Step-I

&

300

Rd2486

F

180.86

200.45

220.37

 

10.83

21.85

Rd2487

F

171.06

190.71

209.52

 

11.49

22.48

Rd2488

F

179.93

197.34

215.24

 

9.68

19.62

 

Mean

 

177.28

196.17

215.04

 

10.66

21.32

 

±SD

 

5.41

4.97

5.43

 

0.92

1.50

Step-I

Confirmation

&
300

Rd2489

F

188.74

205.41

224.83

 

8.83

19.12

Rd2490

F

191.22

210.57

229.70

 

10.12

20.12

Rd2491

F

212.24

230.23

251.49

 

8.48

18.49

 

Mean

 

197.40

215.40

235.34

 

9.14

19.25

 

±SD

 

12.91

13.10

14.20

 

0.86

0.82

Step-II

&
2000

Rd2492

F

187.11

205.49

223.81

 

9.82

19.61

Rd2493

F

202.21

220.31

241.70

 

8.95

19.53

Rd2494

F

215.18

232.84

251.93

 

8.21

17.08

 

Mean

 

201.50

219.55

239.15

 

8.99

18.74

 

±SD

 

14.05

13.69

14.23

 

0.81

1.44

Step-II

Confirmation

&
2000

Rd2495

F

209.20

230.71

258.74

 

10.28

23.68

Rd2496

F

211.92

232.42

255.27

 

9.67

20.46

Rd2497

F

196.16

215.89

239.16

 

10.06

21.92

 

Mean

 

205.76

226.34

251.06

 

10.00

22.02

 

±SD

 

8.42

9.09

10.45

 

0.31

1.61

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off may be considered to be 5000 mg/kg body weight by oral route in the rat.
Executive summary:

The acute oral toxicity of the test item has been tested in accordance with OECD Test Guideline 423, following GLP. 12 female Sprague-Dawley rats divided in 4 groups were administered sequentially with test item previously dissolved in corn oil by oral gavage. The first set of 3 female rats was given a single dose of 300 mg/kg bw. No mortality was observed at this dose level, so a confirmatory set of 3 female rats was treated at the same dose level. No mortality was observed at this dose level. Thus, a third set of 3 female rats was given a single dose of 2000 mg/kg bw. No mortality was observed at this dose level, so finally a confirmatory set of 3 female rats was treated at the same dose level. The body weight evolution of all animals remained normal during the study. No clinical signs of toxicity were observed in Step-I and Step-I confirmation animals dosed with 300 mg/kg bw. In Step-II and Step-II confirmation (animals dosed with 2000 mg/kg bw) no clinical signs were observed on day 1; diarrhea and wet perineum was observed on day 2 and wet perineum was observed on day 3. The observed clinical signs reversed back to normal by day 4 observation. The macroscopic examination of the animals at the end of the study did not reveal treatment related effects. Based on these results, the LD50 of the test item is deterined to be higher than 2000 mg/kg bw. As per OECD TG 423, the LD50 cut-off of the test item may be considered to be 5000 mg/kg bw.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw

The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.
Executive summary:

The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance is greater than 5000 mg/kg bw in rats.
Executive summary:

According to a peer reviewed article, the acute oral LD50 value of the test substance is greater than 5000 mg/kg bw in rats.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
1 680 mg/kg bw
95% CL:
> 1 460 - < 1 900

The acute oral LD50 value in rats was reported as 1.68 g/kg bw.

Interpretation of results:
study cannot be used for classification
Conclusions:
The acute oral LD50 value in rats was reported as 1.68 g/kg bw.
Executive summary:

The acute oral LD50 value in rats was reported as 1.68 g/kg bw (1.46 - 1.90 g/kg).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
4.39 mL/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance is 4.39 mL/kg bw in rats (equivalent to 3789 mg/kg bw).
Executive summary:

According to a peer reviewed article, the acute oral LD50 value of the test substance is 4.39 mL/kg bw in rats (equivalent to 3789 mg/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
3 700 mg/kg bw
Remarks on result:
other: confidence limit: 2300-5100 mg/kg bw

The acute oral LD50 value in rats was reported as 3.7 g/kg bw (2.3 -5.1 g/kg bw)

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value in rats was reported as 3.7 g/kg bw (2.3 -5.1 g/kg bw)
Executive summary:

The acute oral LD50 value in rats was reported as 3.7 g/kg bw (2.3 -5.1 g/kg bw)

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance is greater than 5000 mg/kg bw in rats.
Executive summary:

According to a peer reviewed article, the acute oral LD50 value of the test substance is greater than 5000 mg/kg bw in rats.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw

The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.
Executive summary:

The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1975
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
(Original reference in Japanese language)
Principles of method if other than guideline:
- Principle of test: Standard acute method
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
mouse
Strain:
other: ddN
Sex:
male/female
Details on test animals or test system and environmental conditions:
None
Route of administration:
oral: unspecified
Vehicle:
other: 10% gum arabic-water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 6, 7, 8.5, 10.5, 14, 15, 16.5 and 20 mL/kg bw

DOSAGE PREPARATION (if unusual): Test solution was prepared with 10% gum arabic-water.
Doses:
3000, 3500, 4300, 5300, 7000, 7500, 8300 and 10000 mg/kg bw
No. of animals per sex per dose:
- At 3000 mg/kg bw: 10 males
- At 3500-8300 mg/kg bw: 10 mice/sex
- At 10000 mg/kg bw: 10 females
Control animals:
not specified
Details on study design:
None
Statistics:
No data
Preliminary study:
Not applicable
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
5 600 mg/kg bw
Based on:
test mat.
95% CL:
>= 4 800 - <= 6 500
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
6 600 mg/kg bw
Based on:
test mat.
95% CL:
>= 5 500 - <= 7 900
Mortality:
- Mortalities (male): 0, 10, 0, 30, 60, 90 and 100% at 3000, 3500, 4300, 5300, 7000, 7500 and 8300 mg/kg bw, respectively.
- Mortalities (female): 0, 20, 20, 30, 60, 90 and 100% at 3500, 4300, 5300, 7000, 7500, 8300 and 10000 mg/kg bw, respectively.
Clinical signs:
No data
Body weight:
No data
Gross pathology:
No data
Other findings:
None

None

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The oral LD50 for d-limonene is higher than 5000 mg/kg bw in mice therefore it is not classified according to the CLP Regulation (EC) N° (1272-2008).
Executive summary:

In an acute oral toxicity study, groups (10/sex) of ddN mice were given a single oral dose of d-limonene (50%) in 10% gum-arabic-water at 3000, 3500, 4300, 5300, 7000, 7500, 8300 or 10000 mg/kg bw. Mortalities in males were 0, 10, 0, 30, 60, 90 and 100% at 3000, 3500, 4300, 5300, 7000, 7500 and 8300 mg/kg bw, respectively. Mortalities in females were 0, 20, 20, 30, 60, 90 and 100% at 3500, 4300, 5300, 7000, 7500, 8300 and 10000 mg/kg bw, respectively. The oral LD50 in males and females were 5600 (4800-6500) and 6600 (5500-7900) mg/kg bw, respectively. The oral LD50 for d-limonene is higher than 5000 mg/kg bw in mice therefore it is not classified according to the CLP Regulation (EC) N° (1272-2008).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1975
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
(Original reference in Japanese language)
Principles of method if other than guideline:
- Principle of test: Standard acute method
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Remarks:
(Wistar JCL)
Sex:
male/female
Details on test animals or test system and environmental conditions:
None
Route of administration:
oral: unspecified
Vehicle:
other: 10% gum arabic-water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 3.6, 4.6, 6.0, 7.8, 10.2, 13.2, 17.2, 22.4, 29.2 and 37.8 mL/kg bw

DOSAGE PREPARATION (if unusual): Test solution was prepared with 10% gum arabic-water.
Doses:
1500, 1900, 2500, 3300, 4300, 5600, 7300, 9400, 12200 and 15900 mg/kg bw
No. of animals per sex per dose:
10
Control animals:
not specified
Details on study design:
None
Statistics:
No data
Preliminary study:
Not applicable
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
4 400 mg/kg bw
Based on:
test mat.
95% CL:
>= 3 400 - <= 5 900
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
5 200 mg/kg bw
Based on:
test mat.
95% CL:
>= 3 900 - <= 7 000
Mortality:
- Mortalities (male): 0, 30, 30, 40, 60, 40, 90, 70, 80 and 90% at 1500, 1900, 2500, 3300, 4300, 5600, 7300, 9400, 12200 and 15900 mg/kg bw, respectively.
- Mortalities (female): 0, 20, 50, 40, 50, 60, 60, 90 and 100% at 1500, 1900, 2500, 3300, 4300, 5600, 7300, 9400, 12200 and 15900 mg/kg bw, respectively.
Clinical signs:
No data
Body weight:
No data
Gross pathology:
No data
Other findings:
None

None

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The oral LD50 for d-limonene is higher than 2000 mg/kg bw in rats therefore it is not classified according to the CLP Regulation (EC) N° (1272-2008).
Executive summary:

In an acute oral toxicity study, groups (10/sex) of Wistar JCL rats were given a single oral dose of d-limonene (50%) in 10% gum-arabic-water at 1500, 1900, 2500, 3300, 4300, 5600, 7300, 9400, 12200 and 15900 mg/kg bw. Mortalities in males were 0, 30, 30, 40, 60, 40, 90, 70, 80 and 90% at 1500, 1900, 2500, 3300, 4300, 5600, 7300, 9400, 12200 and 15900 mg/kg bw, respectively. Mortalities in females were 0, 20, 50, 40, 50, 60, 60, 90 and 100% at 1900, 2500, 3300, 4300, 5600, 7300, 9400, 12200 and 15900 mg/kg bw, respectively. The oral LD50 in males and females were 4400 (3400-5900) and 5200 (3900-7000) mg/kg bw, respectively. The oral LD50 for d-limonene is higher than 2000 mg/kg bw in rats therefore it is not classified according to the CLP Regulation (EC) N° (1272-2008).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Method similar to OECD guideline 401, but no information on doses were reported
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
No information on tested doses
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Osborne-Mendel
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: young adults rats
- Fasting period before study: 18 h
- Housing: cages
- Diet (e.g. ad libitum): Food was replaced in cages as soon as animals received their doses.
- Water (e.g. ad libitum): Ad libitum.

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Doses:
Not reported
No. of animals per sex per dose:
5 females and 5 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 2 weeks
- Necropsy of survivors performed: no specified
- Other examinations performed: clinical signs, body weight and time of death
Statistics:
LD50 were computed by the method of Litchfield & Wilcoxon (1949).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
5 075 mg/kg bw
Based on:
test mat.
95% CL:
>= 4 160 - <= 6 190
Mortality:
Death time: 4 hr-5 days
Clinical signs:
Depression, scrawny appearance, porphyrin-like deposits around eyes and nose.
Body weight:
no data
Gross pathology:
no data
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is 5075 mg/kg body weight by oral route in the rat.

Executive summary:

The acute oral toxicity of the test compound was tested following a method similar to OECD Test Guideline 401. Ten young adult Osborne-Mendel rats evenly divided by sex were administered by oral gavage.

Animals were observed for 2 weeks during which time the development of toxic signs was followed and time of death recorded. Animals exposed to terpinyl acetate alpha showed depression, scrawny appearance, and porphyrin-like deposits around eyes and nose. The acute oral LD50 of the test item was determined to be 5075 mg/kg bw (95% confidence limits: 4160 -6190 mg/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
(It is published in a peer reviewed journal but with limitations in design and/or reporting, e.g., it was conducted in mice instead of rats or the lack of data on number of animals used or doses administered)
Qualifier:
no guideline followed
Principles of method if other than guideline:
- Principle of test: The test substance was administered by gavage to groups of 10 male dd-K mice. From the number of deaths during a 7 day observation period after the administration of the drug, the LD50 value was calculated by the Litchfield-Wilcoxon method.
GLP compliance:
not specified
Test type:
standard acute method
Limit test:
no
Species:
mouse
Strain:
other: dd-k
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 25 g
- Diet (e.g. ad libitum): Ad libitum.
- Water (e.g. ad libitum): Ad libitum.

Route of administration:
oral: gavage
Vehicle:
not specified
Doses:
no data
No. of animals per sex per dose:
no data
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 7 days
Statistics:
Litchfiel-Wilcoxon method
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
2 830 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 290 - <= 3 497
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value in mice was reported to be 2830 mg/kg bw (95% CI = 2290-3497).
Executive summary:

Alpha-Terpineol was administered by gavage to groups of 10 male dd-K mice. From the number of deaths during a 7 day observation period after the administration of the test substance, the LD50 value was calculated by the Litchfield-Wilcoxon method. The oral acute toxicity of the test substance was 2830 mg/kg bw (95% CI = 2290 -3497).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Method similar to OECD guideline 401, but no information on doses were reported
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
No information on tested doses
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Osborne-Mendel
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: young adults rats
- Fasting period before study: 18 h
- Housing: cages
- Diet (e.g. ad libitum): Food was replaced in cages as soon as animals received their doses.
- Water (e.g. ad libitum): Ad libitum.

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Doses:
Not reported
No. of animals per sex per dose:
5 females and 5 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 2 weeks
- Necropsy of survivors performed: no specified
- Other examinations performed: clinical signs, body weight and time of death
Statistics:
LD50 were computed by the method of Litchfield & Wilcoxon (1949).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
4 750 mg/kg bw
Based on:
test mat.
95% CL:
>= 3 720 - <= 6 060
Mortality:
Death time: 4 hr-12 days
Clinical signs:
Depression soon after dosing, coma, bloody lacrimation, diarrhoea. Irritable, scrawny appearance for as long as 2 weeks.
Body weight:
no data
Gross pathology:
no data
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is 4750 mg/kg body weight by oral route in the rat.

Executive summary:

The acute oral toxicity of the test compound was tested following a method similar to OECD Test Guideline 401. Ten young adult Osborne-Mendel rats evenly divided by sex were administered by oral gavage.

Animals were observed for 2 weeks during which time the development of toxic signs was followed and time of death recorded. Animals exposed to the test material showed depression soon after dosing, coma, bloody lacrimation and diarrhoea as well as irritable, scrawny appearance for as long as 2 week. The acute oral LD50 of the test item was determined to be 4750 mg/kg bw (95% confidence limits: 3720 -6060 mg/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
3 650 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 710 - <= 4 590
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance was found to be 3650 mg/kg bw in rats.
Executive summary:

According to a peer reviewed article, the acute oral LD50 value of the test substance is 3650 mg/kg bw in rats.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Method similar to OECD guideline 401, but no information on number of animals or doses used were reported
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
No information on number of animals or tested doses
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
mouse
Strain:
not specified
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: no data
- Fasting period before study: No. Mice were treated on full stomachs
- Housing: cages
- Diet (e.g. ad libitum): Food was replaced in cages as soon as animals received their doses.
- Water (e.g. ad libitum): Ad libitum.

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 10-20% (w/v)
Doses:
Not reported
No. of animals per sex per dose:
Not reported
Control animals:
no
Details on study design:
- Duration of observation period following administration: 2 weeks
- Necropsy of survivors performed: no specified
- Other examinations performed: clinical signs, body weight and time of death
Statistics:
LD50 were computed by the method of Litchfield & Wilcoxon (1949).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 250 mg/kg bw
Based on:
test mat.
95% CL:
>= 812 - <= 1 920
Mortality:
Death time: 1 hr-4 days
Clinical signs:
Depression soon after treatment, coma on higher doses
Body weight:
no data
Gross pathology:
no data
Interpretation of results:
other: category 4 (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is 1250 mg/kg body weight by oral route in the mouse.

Executive summary:

The acute oral toxicity of the test compound was tested following a method similar to OECD Test Guideline 401. Groups of mice were administered by oral gavage on full stomachs with test material diluted in corn oil (10 -20% (w/v)). Animals were observed for 2 weeks during which time the development of toxic signs was followed and time of death recorded. Animals exposed to the test material showed depression soon after treatment and coma on higher doses. The acute oral LD50 of the test item was determined to be 1250 mg/kg bw (95% confidence limits: 812 - 1920 mg/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Method similar to OECD guideline 401, but no information on doses were reported
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
No information on tested doses
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Osborne-Mendel
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: young adults rats
- Fasting period before study: 18 h
- Housing: cages
- Diet (e.g. ad libitum): Food was replaced in cages as soon as animals received their doses.
- Water (e.g. ad libitum): Ad libitum.

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Doses:
Not reported
No. of animals per sex per dose:
5 females and 5 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 2 weeks
- Necropsy of survivors performed: no specified
- Other examinations performed: clinical signs, body weight and time of death
Statistics:
LD50 were computed by the method of Litchfield & Wilcoxon (1949).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 820 mg/kg bw
Based on:
test mat.
95% CL:
>= 1 670 - <= 1 980
Mortality:
Death time: 4 hr-8 days
Clinical signs:
Marked depression, some rats in coma for 24 hr, rough fur, wet posterior, porphyrin-like deposit around eyes.
Body weight:
no data
Gross pathology:
no data
Interpretation of results:
other: category 4 (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is 1820 mg/kg body weight by oral route in the rat.

Executive summary:

The acute oral toxicity of the test compound was tested following a method similar to OECD Test Guideline 401. Ten young adult Osborne-Mendel rats evenly divided by sex were administered by oral gavage.

Animals were observed for 2 weeks during which time the development of toxic signs was followed and time of death recorded. Animals exposed to the test material showed marked depression, some rats in coma for 24 hr, rough fur, wet posterior, porphyrin-like deposit around eyes. The acute oral LD50 of the test item was determined to be 1820 mg/kg bw (95% confidence limits: 1670 - 1980 mg/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
not specified
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
3 100 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 600 - <= 3 600
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance is 3.1 g/kg bw in rats.
Executive summary:

According to a peer reviewed article, the acute oral LD50 value of the test substance is 3.1 g/kg bw in rats (95% CL: 2.6 - 3.6 g/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
1 300 mg/kg bw
Based on:
test mat.
95% CL:
>= 840 - <= 2 100
Interpretation of results:
other: category 4 (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance is 1.3 g/kg bw in rats.
Executive summary:

According to a peer reviewed article, the acute oral LD50 value of the test substance is 1.3 g/kg bw in rats (95% CL: 0.84 - 2.1 g/kg bw).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Principles of method if other than guideline:
No data on test method
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Doses:
2050, 2560, 3200, 4000 and 5000 mg/kg bw.
No. of animals per sex per dose:
2 animals for doses of 2050, 2560 and 4000 mg/kg bw
3 animals for dose of 3200 mg/kg bw
5 animals for dose of 5000 mg/kg bw
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Other examinations performed: clinical signs
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
2 050 mg/kg bw
Based on:
test mat.
Mortality:
All deaths occurred between days 1 and 3.
At 2050 and 2560 mg/kg bw, 1/2 animals died; 3/3 animals died at 3200 mg/kg bw; 2/2 animals died at 4000 mg/kg bw and 5/5 animals died at 5000 mg/kg bw.
Clinical signs:
Clinical signs included lethargy, ataxia, tearing, comatose and flaccid (no further details were reported).
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute oral LD50 value of the test substance was found to be 2050 mg/kg bw in rats.
Executive summary:

The acute oral toxicity of the test item was evaluated in rats. The test substance was administered at dose levels of 2050, 2560, 3200, 4000, and 5000 mg/kg body weight. Mortality and clinical observations were analyzed for 14 days after exposure. All deaths occurred between days 1 and 3. At 2050 and 2560 mg/kg bw, 1/2 animals died; 3/3 animals died at 3200 mg/kg bw; 2/2 animals died at 4000 mg/kg bw and 5/5 animals died at 5000 mg/kg bw. Clinical signs included lethargy, ataxia, tearing, comatose and flaccid. The acute oral LD50 value of the test substance was determined to be 2050 mg/kg bw.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Lork’s guidelines (Lorke D., 1983).
Qualifier:
according to guideline
Guideline:
other: Lork’s guidelines (Lorke D., 1983).
GLP compliance:
not specified
Remarks:
this information was not reported
Test type:
other: Lork's method (Lorke D., 1983)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Central Animal House of Urmia University
- Weight at study initiation: 200–230 g
- Fasting period before study: 18–24 h prior to experimentation (food but not water)
- Housing: The animals were housed in polypropylene cages
- Diet (e.g. ad libitum): rodent pellet diet (Dane-Pars Co., Tehran, Iran), ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2ºC
- Humidity (%): 55-60%

- Photoperiod (hrs dark / hrs light): 12 h light/dark cycle



Route of administration:
oral: gavage
Vehicle:
other: Tween 80 (2%)
Details on oral exposure:
VEHICLE
- Amount of vehicle (if gavage): 2 mL

MAXIMUM DOSE VOLUME APPLIED: 2 mL
Doses:
10, 100, 600, 1000, 1600 and 2900 mg/kg
No. of animals per sex per dose:
3
Control animals:
yes
Remarks:
2mL of Tween 80 (2%)
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: not reported.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, food and water consumption, body weight, visual inspection at necropsy
Statistics:
LD50 was calculated based on the pattern of death observed using the Probit-log analysis.
The obtained results of observations and examinations were compared with the control group. p < 0.05 values were considered significant.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 280 mg/kg bw
Based on:
test mat.
Mortality:
There were no treatment-related mortality in animals treated with 1,8-cineole at doses of 10, 100 and 600 mg/kg for 14 d.
Clinical signs:
There were no visible signs of toxicity in animals treated with 1,8-cineole at doses of 10 and 100 mg/kg for 14 d. Even high-dose males and females did not exhibit a significant decrease in food and water consumption for 14 d.
Visible signs of toxicity were reported in the animals at doses of greater than 100 mg/kg. The signs include tremor, convulsion, abnormal gait and ataxia, increased respiration, decreased activity, unresponsiveness to writhing test, and flaccid paralysis that lead to recumbencey.
Body weight:
There was no significant post treatment drop in body weight in both males and females of all groups.
Gross pathology:
Visual inspection on necropsy did not reveal any signs of damage to organs.

Table 1. Acute oral toxicity of 1,8-cineole in rats.

 

 

Effects

Treatment
(g/kg)

D/T

Mortality
latency (h)

Symptoms of toxicity

10

0/3

-

None

100

0/3

-

None

600

0/3

-

Ataxia, hypoactivity, labored
respiration, convulsion,
recovery after 185±5 min

1000

2/3

<6, <15

Ataxia, hypoactivity, labored
respiration, head tics, body
tremor, convulsion, paddling,
flaccid paralysis, recumbencey,
unresponsive to writhing test,
recovery after 24±5 min

1600

1/3

<2, <10

Ataxia, hypoactivity, labored
respiration, head tics, body
tremor, convulsion, flaccid
paralysis, recumbencey,
unresponsive to writhing test,
recovery after 73±5 min

2900

3/3

<2, <3

Ataxia, hypoactivity, labored
respiration, body tremor,
convulsion, paddling, flaccid
paralysis, recumbencey,
unresponsive to writhing test

Symptoms of toxicity are described for a group; D/T, dead/treated rats; latency, time to death after the dose;

None, no toxic symptom during the observation period.

Interpretation of results:
other: Category 4 (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of 1,8-cineole was estimated to be 1280 mg/kg for the oral route in an adult rat.
Executive summary:

The acute oral toxicity of 1,8-cineole in rat was measured in accordance with Lork’s method (Lorke, 1983). 3 rats per group were administered orally (gavage) in a single dose with test material diluted in Tween 80 (2%) at concentrations of 0 (control only solvent), 10, 100, 600, 1000, 1600, and 2900 mg/kg bw to the final volume of 2 mL. Animals were under observation for a period of 14 consecutive days. There were no treatment-related mortality at doses of 10, 100 and 600 mg/kg. Visible signs of toxicity were reported in the animals at doses of greater than 100 mg/kg. The signs include tremor, convulsion, abnormal gait and ataxia, increased respiration, decreased activity, unresponsiveness to writhing test, and flaccid paralysis that lead to recumbencey. No effects were reported in body weight evolution and the visual inspection on necropsy did not reveal any signs of damage to organs. Based on these results, the LD50 of 1,8-cineole was estimated to be 1280 mg/kg.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Key study with Klimisch score 1.

Acute toxicity: via inhalation route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
According to REACH Annex VIII, column 2: In addition to the oral route (Annex VII, 8.5.1.), for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. The information is provided for dermal route.
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
significant methodological deficiencies
Remarks:
Test method was not according to any guideline. The species used was the rabbit. However, the recommended species for the acute inhalation toxicity is the rat. No GLP.
Principles of method if other than guideline:
- Principle of test: Studies on the pharmacological expectorant activity of nutmeg oil and camphene given by inhalation to rabbits arranged for the collection of respiratory tract fluid.
- Short description of test conditions: see below
- Parameters analysed / observed: volumen of respiratory tract fluid, expectorant activity and odor
GLP compliance:
no
Test type:
other: Study of the pharmacological expectorant activity of test substance
Limit test:
no
Species:
rabbit
Strain:
not specified
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: Young
- Weight at study initiation: 2-3 kg
- Housing: They were housed in boarding cages, one animal per cage.
- Diet (e.g. ad libitum): They were fed a standard rabbit ration of mixed natural foods, salts, and vitamins.
Route of administration:
inhalation: vapour
Type of inhalation exposure:
nose only
Vehicle:
other: ethanol
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:
In preparation for the collection of respiratory tract fluid, the rabbits were anesthetized with urethane (ethyl carbamate) given by intraperitoneal injection in a dose of 4.0 to 5.0 mL/kg body weight of a 25% (w/v) solution in distilled water. This produced a light anesthesia lasting over 24 hours with no reported effect on the volume and composition of respiratory tract fluid except that it augmented the concentration of potassium ion.
One arm of a T-shaped cannula was ligated into the trachea, a collecting tube attached to the second arm, and the third arm was connected with a reservoir of conditioned air maintained at 39 ºC and saturated with water vapor. The conditioned air was prepared by mixing laboratory air with steam from a thermostatically controlled water vaporizer. Steam generated by this vaporizer volatilized the test item which was carried to the conditioned air reservoir from which it was inhaled by each anesthetized rabbit.
Respiratory tract fluid was collected for a control period of two to four hours. The collecting tube was then replaced by a cleaned, empty tube, test item added to the vaporizer, and respiratory tract fluid collected for a subsequent period of four to six hours.


Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
>= 4 - <= 6 h
Concentrations:
1, 3, 9, 27, 81, and 243 mg/kg
No. of animals per sex per dose:
4-6 male rabbits per dose
Control animals:
other: The controls were the same animals, before being treated.
Details on study design:
- Duration of observation period following administration: 24 hours, which is a standard method of expressing the volume output of respiratory tract fluid.
Key result
Sex:
male
Dose descriptor:
LC0
Effect level:
>= 243 other: mg/kg
Based on:
test mat.
Exp. duration:
6 h
Mortality:
No animals died during the study.
Clinical signs:
other: Inhalation of camphene by urethanized rabbits in doses 3-27 mg/kg produced significant increase in volume output of respiratory tract fluid, accompanied by decrease in specific gravity and increase in total solids. Dose-dependent increase in volume output

The LC 0 was equal or greater than 243 mg/kg.

Interpretation of results:
study cannot be used for classification
Conclusions:
The LC 0 was equal or greater than 243 mg/kg.
Executive summary:

Camphene was given by inhalation to male rabbits that were anesthetized with urethane (ethyl carbamate) for 24 hours and were arranged for collection of respiratory tract fluid. Test substance was dissolved in 1 mL of ethanol in amounts of 1, 3, 9, 27, 81, and 243 mg. Each dose was given to 4 to 6 rabbits. Respiratory tract fluid was collected for a control period of two to four hours. The collecting tube was then replaced by a cleaned, empty tube, test item added to the vaporizer, and respiratory tract fluid collected for a subsequent period of four to six hours. Steam generated by the water vaporizer volatilized the test item which was carried to the conditioned air reservoir from which it was inhaled by each anesthetized rabbit. Inhalation of camphene by urethanized rabbits in doses 3-27 mg/kg produced significant increase in volume output of respiratory tract fluid, accompanied by decrease in specific gravity and increase in total solids. Dose-dependent increase in volume output of respiratory tract fluid varied with the season. The expectorant action began to disappear following inhalation of a dose of 81 mg/kg, which is well below the level detectable by odour (243 mg/kg) and far below the probably fatal dose. Based on these results, the LC0 was established to be equal or greater than 243 mg/kg.

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Route of administration:
inhalation: vapour
Type of inhalation exposure:
not specified
Vehicle:
not specified
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
Test animals inhaled saturated vapours of alpha pinene from steam distillation of wood.


Analytical verification of test atmosphere concentrations:
not specified
Concentrations:
26 mg/L
No. of animals per sex per dose:
no data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LC50
Effect level:
625 other: mg/L/min
Based on:
test mat.
Remarks on result:
other: Effect level is the LC for the shortest period causing death
Interpretation of results:
study cannot be used for classification
Conclusions:
The LC50 of the test item (for the shortest period causing death) is 625 mg/l/min in rats.
Executive summary:

Alpha pinene was given to rats through inhalation of saturated vapours (26 mg/L of alpha pinene) obtained from steam distillation of wood. The LC50 of the test item (for the shortest period causing death) was found to be 625 mg/l/min.

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 November 2018 - 13 December 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-house bred animals
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 10 weeks
- Weight at study initiation: 256.78 (mean weight of the 5 animals tested)
- Fasting period before study: Animals were fasted overnight (16 to 18 hours) prior to dosing. Water was provided ad libitum during fasting period. Feed was offered 3 to 4 hours after dosing.
- Housing: Maximum of three animals were housed in a standard polypropylene cage (size: L 430 x B 285 x H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube. For range finding study, animals were housed individually after treatment throughout the observation. For main study, during treatment, the animals were housed individually and after patch removal they were housed together. Clean sterilized paddy husk was provided as bedding material. Paper shredding was provided as enrichment.
- Diet (e.g. ad libitum): Ad libitum. Altromin Maintenance Diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG).
- Water (e.g. ad libitum): Ad libitum. Deep bore-well water passed through Reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes.
- Acclimation period: the animals were acclimatized for at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3ºC to 22.8ºC
- Humidity (%): 46 to 66%
- Air changes (per hr): 12-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: dorso-lateral area of the trunk.
- % coverage: at least 10% of the body surface area.
- Type of wrap if used: semi-occlusive dressing (crepe bandage).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test item was washed using distilled water and dried with absorbent cotton.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 200, 1000 and 2000 mg/kg body weight
- Concentration (if solution): N/A
- Constant volume or concentration used: No; Volume applied (mL) = [Dose (mg/kg)/1000 x Body weight (g)/Density (mg/mL)]
- For solids, paste formed: N/A
Duration of exposure:
24 h
Doses:
Range finding study: 200, 1000 and 2000 mg/kg body weight
Main study: 2000 mg/kg body weight
No. of animals per sex per dose:
Range finding study: 1 female per dose
Main study: 2 females per dose
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At each step, the animals were observed for clinical signs of toxicity and mortality at 20 to 30 min, 1 hr (±10 mins), 2 hrs (±10 mins), 4 hrs (±10 mins) and 6 hrs (±10 mins) post dosing on Day 1 and thereafter once daily for clinical signs of toxicity and twice daily for mortality. Individual animal body weight was recorded at receipt, on day 1 before test item application and on day 8 and day 15 during the experimental period.
- Necropsy of survivors performed: yes. At termination, gross pathological findings were recorded and reported.
- Other examinations performed:
Clinical observations: changes in skin, fur, eyes and mucous membranes along with changes in respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during both range finding study and main study.
Clinical signs:
No treatment related clinical signs of toxicity were observed in both range finding study and main study animals.
Body weight:
All animals revealed physiologically normal increase in the body weight in both range finding study and main study.
Gross pathology:
No treatment related gross pathological changes were observed in any of the animals in both range finding study and main study animals.

Table 1:Individual animal clinical signs of toxicity and mortality record

Phase of the Experiment

 

Dose (mg/kg body weight)

Animal No.

Sex

Time of Dosing

(AM)

Clinical Signs of Toxicity and Mortality on Day 1

Clinical Signs of Toxicity and Mortality on days

20-30

mins

1 hr

(±10 mins)

2 hrs

(±10 mins)

4 hrs

(±10 mins)

6 hrs

(±10 mins)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Range Finding Study

200

Rd2498

F

10:58

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

1000

Rd2499

F

10:56

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

2000

Rd2500

F

10:54

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Main Study

2000

Rd2501

F

11:18

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2502

F

11:20

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N: Normal; F: Female; min: minutes; hr/hrs: hour/hours

Table 2: Individual animal body weight (g) and percent change in body weight with respect to day 1

Phase of the Experiment

Dose (mg/kg)

Animal No.

Sex

Body Weight (g) on Days

Percent Change in Body Weight with Respect to Day

1

8

15

1 to 8

1 to 15

Range Finding Study

200

Rd2498

F

250.15

268.38

285.71

7.29

14.22

1000

Rd2499

F

245.11

263.27

284.52

7.41

16.08

2000

Rd2500

F

265.53

283.47

301.23

6.76

13.44

Main Study

2000

Rd2501

F

262.04

281.40

297.94

7.39

13.70

Rd2502

F

251.51

272.18

287.07

8.22

14.14

Mean

 

256.78

276.79

292.51

7.80

13.92

±SD

 

7.45

6.52

7.69

0.59

0.31

Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is higher than 2000 mg/kg body weight by dermal route in rats.

Executive summary:

The acute dermal toxicity of the test item was studied according to OECD Guideline 402 (GLP study). 5 female Sprague-Dawley rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted at doses of 200, 1000 and 2000 mg/kg bw in one animal per dose. Based on the results of this preliminary study two additional rats were tested at a dose of 2000 mg/kg bw. No treatment related clinical signs of toxicity and mortality were observed. The body weight evolution of the animals remained normal during the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related effects. Based on these results, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method.
GLP compliance:
no
Limit test:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Duration of exposure:
No data
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 2 500 mg/kg bw

The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.
Executive summary:

The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method.
GLP compliance:
no
Limit test:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Duration of exposure:
No data
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw

The acute dermal LD50 value in rabbits was reported as greater than 5 g/kg bw.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value in rabbits was reported as greater than 5 g/kg bw.
Executive summary:

The acute dermal LD50 value in rabbits was reported as greater than 5 g/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
not specified
Limit test:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Duration of exposure:
No data
Doses:
No data
No. of animals per sex per dose:
No data
Control animals:
not specified
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 2 500 mg/kg bw

The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.
Executive summary:

The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
No. of animals per sex per dose:
10 animals in total
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
0/10 deaths at the highest dose tested (5000 mg/kg bw)
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance in rabbits was >5 g/kg bw based on 0/10 deaths at that dose.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 2 500 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >2500 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance was >2500 mg/kg bw in rabbits.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Limit test:
yes
Species:
guinea pig
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
2 animals in total
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths ocurred.
Clinical signs:
Not specified
Body weight:
Not specified
Gross pathology:
Not specified
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >2000 mg/kg bw in guinea pigs.
Executive summary:

The acute dermal toxicity of Fenchol was evaluated in 2 guinea pigs. Test substance was administered via dermal application at 2000 mg/kg/body weight. The animals were observed for 14 days. No deaths occurred. Thus, the acute dermal LD50 value of the test item was determined to be greater than 2000 mg/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
No data on test method
GLP compliance:
no
Species:
rabbit
Strain:
not specified
Sex:
not specified
Type of coverage:
not specified
Vehicle:
not specified
Control animals:
not specified
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.
Executive summary:

According to a peer reviewed article, the acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Key study with Klimisch score 1.

Additional information

Acute oral toxicity: Key study. The acute oral toxicity of the test item has been tested in accordance with OECD Test Guideline 423, following GLP. 12 female Sprague-Dawley rats divided in 4 groups were administered sequentially with test item previously dissolved in corn oil by oral gavage. The first set of 3 female rats was given a single dose of 300 mg/kg bw. No mortality was observed at this dose level, so a confirmatory set of 3 female rats was treated at the same dose level. No mortality was observed at this dose level. Thus, a third set of 3 female rats was given a single dose of 2000 mg/kg bw. No mortality was observed at this dose level, so finally a confirmatory set of 3 female rats was treated at the same dose level. The body weight evolution of all animals remained normal during the study.No clinical signs of toxicity were observed in Step-I and Step-I confirmation animals dosed with 300 mg/kg bw.In Step-II and Step-II confirmation (animals dosed with 2000 mg/kg bw) no clinical signs were observed on day 1; diarrhea and wet perineum was observed on day 2 and wet perineum was observed on day 3. The observed clinical signs reversed back to normal by day 4 observation.The macroscopic examination of the animals at the end of the study did not reveal treatment related effects. Based on these results, the LD50 of the test item is deterined to be higher than 2000 mg/kg bw. As per OECD TG 423, the LD50 cut-off of the test item may be considered to be 5000 mg/kg bw.

Acute oral toxicity: Supporting studies. Individual acute toxicities of the main components are available from peer review publications:

Alpha terpineol: Peer reviewed publication (Yamahara J, 1985). The acute oral LD50 value in mice was reported to be 2830 mg/kg bw.

Alpha terpinyl acetate: Peer reviewed publication (Jenner P M, 1964). The LD50 of the test item is 5075 mg/kg body weight by oral route in the rat.

Terpinolene: Peer reviewed publication (Opdyke DLJ, 1976). The acute oral LD50 value of the test substance is 4.39 mL/kg bw in rats (equivalent to 3789 mg/kg bw).

Cineole: Peer reviewed publication (Jalilzadeh-Amin G, 2014). The acute oral LD50 value in rat was reported to be 1280 mg/kg bw.

Isocineole: Peer reviewed publication (fragrance monograph, 1988). The acute oral LD50 value in rats was reported as 3.1 g/kg bw.

L-Limonene: Peer reviewed publication (Opdyke DLJ, 1978). The acute oral LD50 value of the test substance is greater than 5000 mg/kg bw in rats.

D-Limonene: Peer reviewed publication (Opdyke DLJ, 1975). The acute oral LD50 value of the test substance is greater than 5000 mg/kg bw in rats.

D-Limonene: Peer reviewed publication (Tsuji, 1975). The oral LD50 for d-limonene is higher than 2000 mg/kg bw in rats.

D-Limonene: Peer reviewed publication (Tsuji, 1975). The oral LD50 for d-limonene is higher than 5000 mg/kg bw in mice.

Camphene: Peer reviewed publication (Opdyke DLJ, 1975). The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.

Camphene: Peer reviewed handbook (Patty's2001). The acute oral LD50 value in rats was reported as greater than 5 g/kg bw.

Alpha terpinene: Peer reviewed publication (Opdyke DLJ, 1976). The acute oral LD50 value in rats was reported as 1.68 g/kg bw.

Gamma terpinene: Peer reviewed publication (Opdyke DLJ, 1976). The acute oral LD50 value in rats was reported as 3.65 g/kg bw.

Alpha pinene: Peer reviewed publication (Opdyke DLJ, 1978). The acute oral LD50 value in rats was reported as 3.7 g/kg bw.

p-cymene: Peer reviewed publication (Jenner P M, 1964). The LD50 of the test item is 4750 mg/kg body weight by oral route in the rat.

Estragole: Peer reviewed publication (Jenner P M, 1964). The LD50 of the test item is 1820 mg/kg body weight by oral route in the rat and 1250 mg/kg body weight by oral route in the mouse.

Terpinen-4 -ol: Peer reviewed publication (Opdyke DLJ, 1982). The acute oral LD50 value in rats was reported as 1.3 g/kg bw.

Fenchyl alcohol: Peer reviewed publication (Bhatia S P, 2008). The acute oral LD50 value in rats was reported as 2.05 g/kg bw.

Acute inhalation toxicity: Data waiving (study scientifically not necessary): According to REACH Annex VIII, column 2: In addition to the oral route, for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. The information is provided for dermal route.

Acute inhalation toxicity: Supporting studies. Individual acute toxicities of some main components are available from experimental tests.

Camphene: Peer reviewed publication (Boyd EM, 1970). The LC 0 was equal or greater than 243 mg/kg.

Alpha pinene: Peer reviewed publication (Opdyke DLJ, 1978). The LC50 of the test item (for the shortest period causing death) is 625 mg/l/min in rats.

Acute dermal toxicity: Key study. The acute dermal toxicity of the test item was studied according to OECD Guideline 402 (GLP study). 5 female Sprague-Dawley rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted at doses of 200, 1000 and 2000 mg/kg bw in one animal per dose. Based on the results of this preliminary study two additional rats were tested at a dose of 2000 mg/kg bw. No treatment related clinical signs of toxicity and mortality were observed. The body weight evolution of the animals remained normal during the study.The macroscopic examination of the animals at the end of the study did not reveal treatment related effects.Based on these results, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Acute dermal toxicity: Supporting studies. Individual acute toxicities of the main components are available from experimental tests.

Terpinolene: Peer reviewed publication (Opdyke DLJ, 1976). The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Isocineole: Peer reviewed publication (fragrance monograph, 1988).The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

L-Limonene: Peer reviewed publication (Opdyke DLJ, 1978).The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

D-Limonene: Peer reviewed publication (Opdyke DLJ, 1975).The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Camphene: Peer reviewed publication (Opdyke DLJ, 1975). The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.

Camphene: Peer reviewed handbook (Patty's2001). The acute dermal LD50 value in rabbits was reported as greater than 2.5 g/kg bw.

Alpha pinene: Peer reviewed publication (Opdyke DLJ, 1978). The acute dermal LD50 value in rabbits was reported as greater than 5 g/kg bw.

Gamma terpinene: Peer reviewed publication (Opdyke DLJ, 1976). The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Terpinen-4 -ol: Peer reviewed publication (Opdyke DLJ, 1982).The acute dermal LD50 value of the test substance was >2500 mg/kg bw in rabbits.

Fenchyl alcohol: Peer reviewed publication (Bhatia S P, 2008).The acute dermal LD50 value of the test substance was >2000 mg/kg bw in guinea pigs.

Estragole: Peer reviewed publication (Opdyke DLJ, 1976).The acute dermal LD50 value of the test substance was >5000 mg/kg bw in rabbits.

Justification for classification or non-classification

Based on the available data, the substance is not classified for acute toxicity according to CLP Regulation (EC) no. 1272/2008.