Registration Dossier

Administrative data

Description of key information

LD50, acute oral, rat: >2000 mg/kg bw

LD50, acute dermal, rat: >2000 mg/kg bw

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:
November 05, 2012 - December 13, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Qualifier:
according to
Guideline:
other: Japan MAFF 8147
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008
GLP compliance:
yes (incl. certificate)
Remarks:
Bioassay, Labor für biologische Analytik GmbH, INF 515, 69120 Heidelberg
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: approx. 10 - 11 weeks
- Weight at study initiation: 174 - 207 g
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum
- Housing: Single housing in Makrolon cages, type III
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany)
- Water: Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
olive oil
Remarks:
Ph.Eur
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20 g/ml
- Amount of vehicle (if gavage): 10 ml/kg
Doses:
2000 mg/kg
No. of animals per sex per dose:
2 groups of 3 female animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. A check for any dead or moribund animals was made at least once each workday.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
In two out of three animals of the first test group impaired general state, dyspnoea and piloerection were observed at hour 0 and 1 after administration. On study day 2 and 3 all animals showed impaired general state and piloerection. In all animals of this test group reduced feces was noted from study day 1 until study day 3 after administration.
In the second 2000 mg/kg bw dose group all three animals showed impaired general state and piloerection from study day 1 until study day 3. In two of these animals these findings were observed until study day 9. In all animals reduced feces were observed on study day 2 only. Additionally light brown discolored feces were noted in two animals on study day 2 and 3 after administration.
Body weight:
The mean body weight of the test groups increased throughout the study period within the normal range.
Gross pathology:
There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study the median lethal dose of the test article after oral administration was found to be greater than 2000 mg/kg bw in rats.
Executive summary:

In a GLP-compliant acute oral toxicity study performed according to the Acute Toxic Class method (OECD 423), 2000 mg/kg bw of the test item (preparation in olive oil Ph.Eur.) were administered to two test groups of three fasted Wistar rats by gavage. The following test substance-related clinical observations were recorded:

2000 mg/kg (first test group):

- No mortality occurred

- Impaired general state in all animals

- Dyspnoea in two out of three animals

- Piloerection in all animals

- Reduced feces in all animals  

2000 mg/kg (second test group):

- No mortality occurred

- Impaired general state in all animals

- Piloerection in all animals

- Reduced feces in all animals

- Light brown discoloration of feces in two out of three animals

The mean body weight increased within the normal range throughout the study period. There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period. The acute oral LD50 in rats was calculated to be > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw

Acute toxicity: via inhalation route

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:
February 10, 2012 - March 03, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008
Qualifier:
according to
Guideline:
other: Japan MAFF 8147
GLP compliance:
yes (incl. certificate)
Remarks:
Bioassay, Labor für biologische Analytik GmbH, 69120 Heidelberg
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: male animals approx. 8 weeks, female animals approx. 12 weeks
- Weight at study initiation: approx. 230 g (males) and approx. 200 g (females)
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum
- Housing: Single housing in Makrolon cages, type III
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany)
- Water: Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Vehicle:
olive oil
Remarks:
Ph.Eur
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal and dorso-lateral parts of the trunk
- % coverage: at least 10 % of total body surface area
- Type of wrap if used: The test item was covered with an air-permeable dressing (4 layers of absorbent gauze (Ph. Eur. supplied by Lohmann GmbH & Co., KG) and stretch bandage (Fixomull® Stretch (adhesive fleece) supplied by Beiersdorf AG).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with warm water
- Time after start of exposure: 24 h

VEHICLE
- Amount(s) applied (volume or weight with unit): 3.33 ml/kg body weight
Duration of exposure:
24 h
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: A check for any dead or moribund animals was made at least once each workday, these records are archived by Bioassay. Individual body weights were determined shortly before administration (day 0), weekly thereafter and on the last day of observation. Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals.
- Necropsy of survivors performed: yes
- Other examinations performed: Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation. The evaluation of skin reactions was performed according to Draize, J.H. (1959).
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination. No local effects were observed.
Body weight:
The mean body weight of the male animals increased within the normal range throughout the study period with the exception of one male which showed stagnation of body weight during the second week. The mean body weight of the female animals did not adequately increase during the first post-exposure observation week probably due to the bandage procedure, but increased during the second week within the normal range.
Gross pathology:
No macroscopic pathologic abnormalities were noted in the animals (5 males and 5 females) examined on the last day of observation.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study the median lethal dose (LD50) of the test article after dermal application was found to be greater than 2000 mg/kg bw in male and female rats.
Executive summary:

In a GLP-compliant acute dermal toxicity study (Limit Test) following OECD guideline 402, young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the test article (as suspension in olive oil Ph.Eur) to the clipped skin (dorsal and dorso-lateral parts of the trunk) and covered by semi-occlusive dressing for 24 hours. The application area comprised at least 10% of the total body surface area. The animals were observed for 14 days. No mortality occurred. No signs of systemic toxicity or skin effects and no macroscopic pathologic abnormalities were noted in the animals examined at the end of the study. The mean body weight of the female animals did not adequately increase during the first post-exposure observation week, probably due to the bandage procedure, but increased during the second week within the normal range. The mean body weight of the male animals increased within the normal range throughout the study period, with the exception of one male which showed stagnation of body weight during the second week. Accordingly, the acute dermal median lethal dose (LD50) in rats was determined to be greater than 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw

Additional information

Acute oral toxicity

An acute oral toxicity study following OECD TG 423 and in compliance with GLP was performed to characterize the acute toxicity of the test material (Bioassay, 2013). The test material was administered to two test groups each consisting of three female Wistar rats at the limit dose of 2000 mg/kg body weight. No mortality occurred within the 14 day observation period. Clinical signs observed were impaired general state, dyspnoea and piloerection. In addition, in all animals reduced feces was reported and 2 animals of the second group displayed light brown discolored feces. The mean body weight of the test groups increased throughout the study period within the normal range. There were no macroscopic findings noted during necropsy at the end of the observation period. In conclusion, under the conditions of this study, the median lethal dose in rats after oral administration was found to be greater than 2000 mg/kg body weight.

In a supporting study performed by IBT in 1972, groups of 4 rats (2 male and 2 female) were treated with the test article by gavage at dose levels of 6834, 10250, 15380 and 23070 mg/kg. No mortality was observed in the lowest dose group, whereas 2/4 animals died in the two mid dose groups and all animals died in the highest dose group. Clinical signs included hypoactivity, ruffled fur, muscular weakness, diarrhea, diuresis, hemorrhagic rhinitis and labored breathing. Based on these results, the LD50 was set at 12560 mg/kg. Considering the history of IBT (reporting of fake data), the reliability of this study is questionable and an accurate Klimisch rating is impossible. Consequently, the study is rated with Klimisch 4. However, since falsified data was predominantly reported for studies with repeated exposure, the data from this IBT study was taken into account in a weight of evidence approach. The IBT data supports the findings observed in the reliable key study. It is concluded that the test article is not toxic after oral ingestion and does not require classification.

Acute dermal toxicity

In a GLP-compliant acute dermal toxicity study (Limit Test) following OECD guideline 402, young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the test article (as suspension in olive oil Ph.Eur) to the clipped skin (dorsal and dorso-lateral parts of the trunk) and covered by semi-occlusive dressing for 24 hours. The application area comprised at least 10% of the total body surface area. No mortality occurred during the observation period of 14 days. No signs of systemic toxicity or skin effects and no macroscopic pathologic abnormalities were noted in the animals examined at the end of the study. The mean body weight of the female animals did not adequately increase during the first post-exposure observation week, probably due to the bandage procedure, but increased during the second week within the normal range. The mean body weight of the male animals increased within the normal range throughout the study period, with the exception of one male which showed stagnation of body weight during the second week. In conclusion, under the conditions of this study the median lethal dose (LD50) in rats after dermal application was found to be greater than 2000 mg/kg body weight.

Acute inhalation toxicity

No reliable key study is available. In the study performed by IBT, 5 male and female rats were exposed to an atmosphere saturated with dusts of the test material for 4 consecutive hours. No mortality occurred. The measured concentration of test material was given as 0.4 mg/l. Similar as above, the reliability of this study is questionable. However, since 2 routs are covered (oral and dermal), no further testing is required according to REACh).

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. Based on the data, classification for acute toxicity is not warranted under Regulation (EC) No.1272/2008.