Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Acute oral toxicity: Key study. Method according to OECD guideline 423, GLP study. The LD50 of the test item is lower than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off may be considered to be 500 mg/kg body weight by oral route in the rat.

Acute inhalation toxicity: Data waiving (study scientifically not necessary/other information available): According to REACH Annex VIII, column 2, in addition to the oral route, information from at least other route must be provided depending on the nature of the substance and the likely route of human exposure. Due to the low vapour pressure of the test material the dermal route is the second route of exposure selected.

Acute dermal toxicity: Key study. Method according to OECD guideline 402, GLP study. The LD50 of the test item is 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
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 December 2017 - 25 December 2017
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:
Wistar
Remarks:
RccHan: WIST
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation, India
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: 166.9 to 188.5 g
- Fasting period before study: overnight prior to dosing until three hours post-dosing
- Housing: 3 females in polypropylene rat cages covered with stainless steel grid top. Autoclaved clean rice husk was used as the bedding material. Wooden blocks were provided as enrichment material.
- Diet (e.g. ad libitum): Ad libitum. Teklad Certified Global High Fiber Rat/Mice Feed manufactured by Envigo, USA. The quality of feed is regularly monitored at Jai Research Foundation.
- Water (e.g. ad libitum): Ad libitum. UV sterilized water filtered through Reverse Osmosis water filtration system. The quality of water is regularly monitored at Jai Research Foundation.
- Acclimation period: 6 to 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23°C
- Humidity (%): 49 to 66%
- Air changes (per hr): At least 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light (06.00 to 18.00 h) and 12 hours darkness.
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: recommended by OECD 423 guideline

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: without preliminary information, the selected starting dose is 300 mg/kg body weight.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
Set I (300 mg/kg bw): 3 animals; Set II (300 mg/kg bw): 3 animals; Set III (2000 mg/kg bw): 3 animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: toxicity and mortality at 30 min, 1h, 2 h, 3h, 4h, 6 h after adminitration. Morbidity and mortality twice a day and clinical sings daily during 14 days. Weighing= D0 (prior to dosing) then on D7, and D14.
- Necropsy of survivors performed: yes. At termination, macroscopic observation was observed.
- Other examinations performed:
Clinical observations: evaluation of skin and fur, eyes and mucous membranes, respiratory and circulatory effects, autonomic effects and central nervous system effects, behavioural pattern, somatomotor activity and observation of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
ca. 500 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed in set I and set II treated at the dose level of 300 mg/kg bw.
All the rats were found dead in set III treated at the dose level of 2000 mg/kg bw.
Clinical signs:
other: No clinical sign was observed in rats treated with 300 mg/kg bw. Clinical sign like lethargy was observed in all the rats treated with 2000 mg/kg bw.
Gross pathology:
External examination of found dead and terminally sacrificed rats did not reveal any abnormality.
Visceral examination of found dead rats revealed pale liver (rats treated with 2000 mg/kg bw) whereas terminally sacrificed rats did not reveal any lesion.
Lesion observed in the found dead rats could be correlated with the test item used in the present study.

Table 1: Mortality

Dose

(mg/kg body weight)

Set

Number of Rats Used

Mortality after Dosing

At Hour

On Day

0.5 – 4

6

1

2

3

4 - 7

8 - 14

300

I

3

0

0

0

0

0

0

0

II

3

0

0

0

0

0

0

0

2000

III

3

0

0

0

0

0

0

3

Table 2: Individual and Mean Body Weight (g) and Body Weight Change (%)

Dose

(mg/kg body weight)

Rat N°

Date and Time of Dosing

Volume of Test Item Administered (mL)

Body Weight (g) on Day

Percent Body Weight Change on Day

0

7

14

At Death

7

14

300

1

December 07, 2017

and

10:00 am

1.67

166.9

170.4

207.1

-

2.1

24.1

2

1.71

170.8

181.3

193.2

-

6.1

13.1

3

1.73

173.3

185.6

200.5

-

7.1

15.7

Mean

170.3

179.1

200.3

-

5.1

17.6

Standard Deviation

(±)

3.2

7.8

7.0

-

2.6

5.7

4

December 11, 2017

and

10:30 am

1.88

187.7

199.3

209.4

-

6.2

11.6

5

1.89

188.5

191.9

196.3

-

1.8

4.1

6

1.86

186.4

195.9

204.5

-

5.1

9.7

Mean

187.5

195.7

203.4

-

4.4

8.5

Standard Deviation

(±)

1.1

3.7

6.6

-

2.3

3.9

2000

7

December 13, 2017

and

10:30 am

1.82

181.7

154.9

-

134.1

-14.7

-

8

1.85

185.1

155.7

-

132.3

-15.9

-

9

1.80

180.2

151.5

-

130.6

-15.9

-

Mean

182.3

154.0

-

-

-15.5

-

Standard Deviation

(±)

2.5

2.2

-

-

0.7

-

Key: Day 0 = Before dosing , - = Not applicable

Table 3: Individual clinical observations

Set I                                                                                                                          

Dose

(mg/kg body weight)

Rat N°

Clinical Signs Observed Post-dosing

At Hour (Day 0)

0.5

1

2

3

4

6

300

1

1

1

1

1

1

1

2

1

1

1

1

1

1

3

1

1

1

1

1

1

 

Dose

(mg/kg body weight)

Rat N°

Clinical Signs Observed Post-dosing

On Days

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300

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

3

1

1

1

1

1

1

1

1

1

1

1

1

1

1

 

Set II                                                                                                                           

Dose

(mg/kg body weight)

Rat N°

Clinical Signs Observed Post-dosing

At Hour (Day 0)

0.5

1

2

3

4

6

300

4

1

1

1

1

1

1

5

1

1

1

1

1

1

6

1

1

1

1

1

1

 

Dose

(mg/kg body weight)

Rat N°

Clinical Signs Observed Post-dosing

On Days

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300

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

Set III                                                                                                                                                                   

Dose

(mg/kg body weight)

Rat N°

Clinical Signs Observed Post-dosing

At Hour (Day 0)

0.5

1

2

3

4

6

2000

7

1

1

1

1

1

1

8

1

1

1

1

1

1

9

1

1

1

1

1

1

 

Dose

(mg/kg body weight)

Rat N°

Clinical Signs Observed Post-dosing

On Days

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

7

1

1

1

1

1

1

1

11

11

11, 2

-

-

-

-

8

1

1

1

1

1

11

11

11

11

2

-

-

-

-

9

1

1

1

1

1

1

1

11

11

11, 2

-

-

-

-

Clinical Signs: 1 = Normal, 2 = Dead, 11 = Lethargy

Note: Day 0 = Day of dosing, - = Not Applicable

Table 4: Individual necropsy findings

Dose

(mg/kg body weight)

Set

Rat

Mode of Death

External

Internal

300

I

1

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

II

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

2000

III

7

Found dead

No abnormality detected

Liver:Pale (1+)

8

Found dead

No abnormality detected

Liver:Pale (2+)

9

Found dead

No abnormality detected

Liver:Pale (+)

Key: + = Minimal, 1+ = Mild, 2+ = Moderate

Interpretation of results:
other: Category 4 (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is lower than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off may be considered to be 500 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. 9 female Wistar rats divided in 3 groups were administered sequentially with test item by oral gavage. The first set of 3 female rats was given a single dose of 300 mg/kg body weight. No mortality was observed at this dose level, so the second set of 3 female rats was treated at the same dose level. No mortality was observed at this dose level, so the third set of 3 female rats was treated at the higher dose level of 2000 mg/kg bw. All the rats were found dead at this dose level. Clinical sign like lethargy was observed from day 6 until day of death and also decrease in body weight was observed on day 7 in rats treated at this high dose. External examination of found dead and terminally sacrificed rats did not reveal any abnormality. Visceral examination of found dead rats revealed pale liver whereas terminally sacrificed rats did not reveal any lesion. Based on these results, the LD50 of the test item is lower than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be 500 mg/kg body weight by oral route in the rat.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
500 mg/kg bw
Quality of whole database:
Key study with Klimisch score = 1

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
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
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
According to REACH Annex VIII, column 2, in addition to the oral route information from at least other route must be provided depending on the nature of the substance and the likely route of human exposure. Due to the low vapour pressure of the test material the dermal route is the second route of exposure selected.
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 December 2017 - 01 January 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:
Wistar
Remarks:
RccHan: WIST
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation, India
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: 219.8 to 226.0 g
- Fasting period before study: No
- Housing: 3 females in polypropylene rat cages covered with stainless steel grid top. Autoclaved clean rice husk was used as the bedding material. Wooden blocks were provided as enrichment material.
- Diet (e.g. ad libitum): Ad libitum. Teklad Certified Global High Fiber Rat/Mice Feed manufactured by Envigo, USA. The quality of feed is regularly monitored at Jai Research Foundation.
- Water (e.g. ad libitum): Ad libitum. UV sterilized water filtered through Reverse Osmosis water filtration system. The quality of water is regularly monitored at Jai Research Foundation.
- Acclimation period: 6 to 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23°C
- Humidity (%): 49 to 66%
- Air changes (per hr): At least 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light (06.00 to 18.00 h) and 12 hours darkness.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure:
- % coverage: at least 10%
- Type of wrap if used: porous gauze dressing (not more than 8 ply) and a non-irritating tape (Medi tape 330 hypo-allergenic surgical tape)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cotton soaked in RO water (freshly collected)
- 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: N/A
- For solids, paste formed: no
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: toxicity and mortality at 0.5, 2, 4 and 6 h post-dermal application on the day of dosing (day 0). Morbidity and mortality twice a day. Erythema and oedema were recorded at 24 h, 48 h and 72 h post patch removal. The clinical signs were recorded once a day. Individual body weight was recorded prior to dermal application on day 0 and on days 7 and 14 post dermal application.
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical observations: evaluation of skin and fur, eyes and mucous membranes, respiratory and circulatory effects, autonomic effects and central nervous system effects, behavioural pattern, somatomotor activity and observation of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study.
Clinical signs:
other: No clinical signs related to the administration of the test item were observed. Erythema and oedema was not evident at 24, 48 and 72 h post patch removal in all the rats.
Gross pathology:
The macroscopic examinations of the animals at the end of the study did not reveal treatment-related changes.

Table 1: Individual and Mean Body Weight (g) and Body Weight Change (%)

 

Dose

(mg/kg body weight)

Date and

Time of Dosing

Animal N°

Volume

 of

test itemApplied (mL)

Body Weights (g) on Day

Percent Body Weight Change on Day

0

7

14

7

14

200

December 07, 2017 and

10:15 am

1

43.96

219.8

235.0

250.3

6.9

13.9

1000

December 11, 2017 and

10:20 am

2

226.0

226.0

232.9

236.3

3.1

4.6

2000

December 13, 2017 and

10:35 am

3

451.8

225.9

226.9

232.0

0.4

2.7

December 15, 2017 and

10:40 am

4

447.4

223.7

231.7

240.1

3.6

7.3

December 18, 2017 and

10:45 am

5

441.0

220.5

210.0

212.1

-4.8

-3.8

Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Remarks:
EU criteria.
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 402 (GLP study). 5 female Wistar rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted sequentially at doses of 200, 1000 and 2000 mg/kg bw in one animal per dose. Based on the results of range finding two additional rats were tested sequentially at a dose of 2000 mg/kg bw. At the end of the 24 h exposure period, the residual test item was removed with cotton soaked in RO water.The skin reactions of each rats were observed at 24, 48 and 72 h post patch removal.The rats were observed for a period of 14 days. There were no treatment-related mortality, clinical signs or necropsy findings recorded. Body weight gain was normal for all rats except for one rat whose body weight decreased on days 7 and 14 compared to day 0 but increased from day 7 to 14. Erythema and oedema was not evident at 24, 48 and 72 h post patch removal in all the rats. Based on the results of the study, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

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. 9 female Wistar rats divided in 3 groups were administered sequentially with test item by oral gavage.The first set of 3 female rats was given a single dose of 300 mg/kg body weight. No mortality was observed at this dose level, so the second set of 3 female rats was treated at the same dose level. No mortality was observed at this dose level, so the third set of 3 female rats was treated at the higher dose level of 2000 mg/kg bw. All the rats were found dead at this dose level. Clinical sign like lethargy was observed from day 6 until day of death and also decrease in body weight was observed on day 7 in rats treated at this high dose. External examination of found dead and terminally sacrificed rats did not reveal any abnormality. Visceral examination of found dead rats revealed pale liver whereas terminally sacrificed rats did not reveal any lesion. Based on these results, the LD50 of the test item is lower than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be 500 mg/kg body weight by oral route in the rat.

Acute inhalation toxicity: Data waiving (study scientifically not necessary/other information available): According to REACH Annex VIII, column 2, in addition to the oral route, information from at least other route must be provided depending on the nature of the substance and the likely route of human exposure. Due to the low vapour pressure of the test material the dermal route is the second route of exposure selected.

Acute dermal toxicity: Key study. The acute dermal toxicity of the test item was studied according to OECD 402 (GLP study). 5 female Wistar rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted sequentially at doses of 200, 1000 and 2000 mg/kg bw in one animal per dose. Based on the results of range finding two additional rats were tested sequentially at a dose of 2000 mg/kg bw. At the end of the 24 h exposure period, the residual test item was removed with cotton soaked in RO water.The skin reactions of each rats were observed at 24, 48 and 72 h post patch removal.The rats were observed for a period of 14 days. There were no treatment-related mortality, clinical signs or necropsy findings recorded. Body weight gain was normal for all rats except for one rat whose body weight decreased on days 7 and 14 compared to day 0 but increased from day 7 to 14. Erythema and oedema was not evident at 24, 48 and 72 h post patch removal in all the rats. Based on the results of the study, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Justification for classification or non-classification

Based on the available data, the substance is classified as category 4 for acute toxicity (oral) according to CLP Regulation (EC) no. 1272/2008.

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