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Description of key information

Acute toxicity: oral route.

- Key study: OECD 420 and EU Method B.1 bis. GLP study. The oral lethal dose 50 (LD50) of the test item in rats is greater than 2000 mg/kg b.w.

Acute toxicity: dermal route

- Key study: OECD 402 and EU B.3. GLP study. The dermal LD50 of the test item in rats is greater than 2000 mg/kg bw.

Acute toxicity: inhalation route

- Data waiving (other justification): According to Regulation (EC) No. 1907/2006, Annex VIII, 8.5.3, column 2, the acute toxicity by inhalation route is not required, as both oral and dermal studies are available.

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:
From 8 March 2016 to 27 April 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Purity: 92%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, humidity should be avoid.


Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 11 - 12 week old.
- Weight at study initiation: For the preliminary study 236-245 g. For the main study an average body weight of 231 g.
- Fasting period before study: 19h
- Housing:plastic cages covered with wire bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height). In the preliminary experiment, the animals were caged individually. In the main experiment, there were four animals in one cage.UV-sterilized wood shavings were used as bedding
- Diet: ad libitum
- Water: ad libitum
- Acclimation period:The animals were quarantined and observed daily for 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 – 25 °C
- Humidity (%): 30 – 51%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES:
Preliminary experiment: The animal treated with 300 mg/kg bw from 30.03.2016 to 13.04.2016. The animal treated with 2000 mg/kg bw from 05.04.2016 to 19.04.2016.
Main experiment: From 08.04.2016 to 22.04.2016.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle:
For the preliminary experiment - 60 mg test item/1 ml oil suspension in one animal and 400 mg/1 ml oil suspension in a second animal.
For the main experiment: 400 mg/1 ml oil suspension
- Amount of vehicle (if gavage): 0.5 mL/100 g b.w. in the preliminary and main experiment
- Justification for choice of vehicle: Due to the low water solubility of the test item.
Doses:
Preliminary experiment: 300 mg/kg b.w.and 2000 mg/kg b.w.
Main experiment: 2000 mg/kg b.w.
No. of animals per sex per dose:
Preliminary experiment: 1 animal per sex per dose
Main experiment: 4 animals per sex per dose
The animal from the preliminary experiment which had received the dose of 2000 mg/kg b.w. was included in the main experiment.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The observation of all animals for morbidity and mortality was conducted twice a day or once a day (on days off) during the 14-day experiment. Body weights of the animals were determined on days 0, 7, and 14.
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical signs: evaluation of general condition of the animals and detailed clinical observations (locomotor system, behaviour, reactions to stimuli, skin and hair, eyes and eyelides, respiratory system, urinary system, digestive system, reproductive system).
Gross examinations: observation of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with their content.
Statistics:
Not performed
Preliminary study:
No signs of toxicity were stated on animals treated with 300 mg/kg b.w.and 2000 mg/kg b.w. Both animals survived the experiment. Body weight gain was observed in both animals but gross examinations did not reveal any pathological changes. According to these results, in the main experiment was administered a dose of 2000 mg/kg b.w.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The animals survived the experiment.
Clinical signs:
No signs of toxicity were stated.
Body weight:
During the second week of the experiment, body weight loss in two females was observed. During the 14-day experiment body weights of all animals increased.
Gross pathology:
Gross examinations did not reveal any pathological changes in the examined animals.

Body weights of the animals (g) – overall list.

Dose

(mg/kg b.w.)

Animal

number

Day of the experiment/

Body weight (g)

Body weight

gain(g)

(From 0 to 14)

0

7

14

300

1*

236

261

264

28

  

2000

1*

245

266

269

24

  

2000

2

231

253

253

22

 

2000

3

227

252

247

20

  

2000

4

238

265

259

21

  

2000

5

228

259

264

36

* the female from the preliminary experiment.

Interpretation of results:
GHS criteria not met
Conclusions:
The median lethal dose (LD50) of the test item on female rats is greater than 2000 mg/kg b.w.
Executive summary:

The acute oral toxicity of the test item was determined on female Wistar rats in accordance to the OECD Guideline 420 with GLP. A preliminary experiment was conducted in which the test item at a single dose of 300 mg/kg b.w. was administered to one animal. Since no evident toxicity was observed in dose 300 mg/kg b.w., the test item at a single dose of 2000 mg/kg b.w. was administered to the second animal. No signs of toxicity or mortality were observed on both animals. On the grounds of the preliminary experiment results, four animals used in the main experiment were given the test item at a dose of 2000 mg/kg b.w. The test item in the form of an oil suspension in a volume of 0.5 mL/100 g b.w. was administered with a metal stomach tube. After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment and body weights of the animals were determined on day 0, 7 and 14. During the 14-day experiment body weights of all animals increased. No signs of toxicity were stated and the animals survived the experiment. Gross examinations did not reveal any pathological changes in the examined animals. On the grounds of the study results LD50 of the test item is greater than 2000 mg/kg b.w.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has a Klimisch score of 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
Study scientifically not necessary / other information available: In accordance with column 2 of REACH Annex VIII, the study does not need to be conducted because exposure of humans via inhalation is likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols or droplets of an inhalable size.
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:
From 8 March 2016 to 21 April 2016
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
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
- Analytical purity: 92%

Species:
rat
Strain:
Wistar
Remarks:
Cmdb: WI; outbred
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 9-week-old males and 15-week-old females
- Weight at study initiation: Males average body weight was 272.6 g, females average body weight was 225.2 g
- Housing: The animals were housed in plastic cages covered with wire bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height). After the application of the test item, each animal was housed individually. After the removal of the test item from the animals’ skin, there were five rats in one cage. Each sex was kept separately.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: The animals were quarantined and observed daily for 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 – 23 °C
- Humidity (%): 30 – 50%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES: From: 30.03.2016 To:13.04.2016
Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal area of the trunks, 30-45cm2 was shaved, patch size 6 x 6 cm
- % coverage: he area of skin treated with the test item was 10% of the body surface area
- Type of wrap if used: The patches were laid on the prepared skin and covered with PVC foil (no contact with skin). An elastic bandage and a sticking plaster were used to make a circular protecting band.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Test item was removed using water
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg b.w.

VEHICLE
- Amount(s) applied (volume or weight with unit): The test item, in form of powder was moistened of a few drops of water (to provide better contact with the skin).
Duration of exposure:
24h
Doses:
2000 mg/kg b.w.
No. of animals per sex per dose:
5 animals/sex/dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: evaluation of general condition twice a day or once a day (on days off) during the 14-day experiment. Body weights of the animals were determined on days 0 (directly before the application of the test item), 7, and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs - Detailed clinical observations were made on the application day (day 0) at hourly intervals up to 5 hours. From the 1st to the 14th day of the experiment, the detailed clinical observations were performed once a day.
Preliminary study:
Not carried out
Key result
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals survived the experiment.
Clinical signs:
The animals did not exhibit any general clinical signs. No pathological changes on the treated skin were noticed.
Body weight:
During the first week of the experiment, body weight loss in two females was observed. During the 14-day experiment, body weight loss in two females was observed. During the 14-day experiment body weights of the remaining animals increased.
Gross pathology:
The gross examination did not reveal any pathological changes in the examined animals.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal LD50 of the test item was > 2000 mg/kg bw in rats.
Executive summary:

The acute dermal toxicity of the test item was determined in accordance to the OECD Guideline 402 with GLP. The test item at a single dose of 2000 mg/kg b.w. was applied to the shaved dorsal area of the trunks of 5 males and 5 females rats for 24 hours. After the application of the test item, the animals were observed for 14 days. General and detailed clinical observations of all animals were performed daily during the entire experiment. Body weights of the animals were determined on days 0 (directly before the application of the test item), 7, and 14. After the 14-day observation period, the animals were euthanized, dissected, and subjected to detailed gross examinations. The animals did not exhibit any general clinical signs. No pathological changes on the treated skin were noticed. All animals survived the experiment. These results indicate that the acute dermal LD50 of the test item is > 2000 mg/kg bw in rats.

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

Additional information

Acute toxicity : oral

The acute oral toxicity of the test item was determined on female Wistar rats in accordance to the OECD Guideline 420 with GLP. A preliminary experiment was conducted in which the test item at a single dose of 300 mg/kg b.w. was administered to one animal. Since no evident toxicity was observed in dose 300 mg/kg b.w., the test item at a single dose of 2000 mg/kg b.w. was administered to the second animal. No signs of toxicity or mortality were observed on both animals. On the grounds of the preliminary experiment results, four animals used in the main experiment were given the test item at a dose of 2000 mg/kg b.w. After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment and body weights of the animals were determined on day 0, 7 and 14. During the 14-day experiment body weights of all animals increased. No signs of toxicity were stated and the animals survived the experiment. Gross examinations did not reveal any pathological changes in the examined animals. On the grounds of the study results LD50 of the test item is greater than 2000 mg/kg b.w.

Acute toxicity : dermal

The acute dermal toxicity of the test item was determined in accordance to the OECD Guideline 402 with GLP. The test item at a single dose of 2000 mg/kg b.w. was applied to the shaved dorsal area of the trunks of 5 males and 5 females rats for 24 hours. After the application of the test item, the animals were observed for 14 days. General and detailed clinical observations of all animals were performed daily during the entire experiment. Body weights of the animals were determined on days 0 (directly before the application of the test item), 7, and 14. After the 14-day observation period, the animals were euthanized, dissected, and subjected to detailed gross examinations. The animals did not exhibit any general clinical signs. No pathological changes on the treated skin were noticed. All animals survived the experiment. These results indicate that the acute dermal LD50 of the test item is > 2000 mg/kg bw in rats.

Justification for classification or non-classification

Based on the available information (oral and dermal LD50 higher than 2000 mg/kg bw), the substance is not classified for acute toxicity according to CLP Regulation (EC) no. 1272/2008.