Registration Dossier

Administrative data

Description of key information

Acute toxicity was studied in rodents after oral and dermal administration. The oral route was chosen as most relevant route of exposure. The dermal study was initiated based on data obtained in the OECD 429 dose range finiding assay. Here, the test material caused severe systemic toxicity in mice after dermal administration using DMF as vehicle at concentrations down to 10%, significant body weight loss down to 2.5% and local irritation at lower concentrations down to 0.5 %. Therefore, an additional dermal toxicity study in rats was initiated to ensure occupational safety. The following results have been obtained:

OECD 423: LD50 > 2000 mg/kg bw

OECD 402: LD50 > 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:
August 19, 2016 - March 7, 2017
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)
Version / remarks:
adopted on December 17, 2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Version / remarks:
Council Regulation (EC) No. 440/2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006 of the European Parliament and the council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final preparation of a solid: The test item preparation was made directly before administration. Appropriate amounts of the test item were suspended in the vehicle using a spatula, a mini shaker (Vortex Genie 2®, Scientific Industries Inc, New York, USA), Ultra-Turrax device (Ultra-Turrax T25, IKA®-Werke GmbH & Co. KG, Staufen, Germany) and a magnetic stirrer. The test item preparation was administered within less than 1 hour after preparation. The stability of the test item in the vehicle was not investigated.
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at start of study: 9 weeks
- Weight at study initiation: The mean initial body weight at the start of study was 167 g (range from 157 to 182 g).
- Fasting period before study: about 17 to 20 hours before start of treatment until 4 hours after administration
- Housing: separately in type III Makrolon cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.4 – 23.3°C
- Humidity (%): 40.9 – 67.8%
- Photoperiod (hrs dark / hrs light): 12 hour light - 12 hour dark regime

IN-LIFE DATES: From: day 1 To: day 15
Route of administration:
oral: gavage
Vehicle:
methylcellulose
Remarks:
Methocel K4M Premium solution
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 5, 30 and 200 g/L
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: well tolerated and established standard vehicle

MAXIMUM DOSE VOLUME APPLIED: 1 mL/100 g
Doses:
50, 300 and 2000 mg/kg bw
No. of animals per sex per dose:
6 (f)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Frequency of weighing: day 1, 2, 4, 6, 8, 11, 13 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No mortality occurred during the course of this study.
Clinical signs:
No clinical signs of toxicity were observed.
Body weight:
The body weight development was inconspicuous throughout the study.
Gross pathology:
The gross pathological examination revealed no organ alterations.
Interpretation of results:
GHS criteria not met
Conclusions:
The test item has no acute toxic potential under the conditions of the present study, and the LD50 value is higher than 2000 mg/kg bw after single oral administration in female rats.
Executive summary:

Objective

The objective of the present study was to identify potential toxic effects of the test item after single oral administration to rats in a stepwise procedure.

Study Design

The study was started with 50 mg/kg bw in 3 female rats, continued with further 3 females treated with 50 mg/kg. Due to the fact, that no mortality was seen after treatment with 50 mg/kg bw, 6 further females were treated with 300 mg/kg bw, where no mortality was seen as well. Therefore, the study was continued with 6 females treated with 2000 mg/kg bw.

Mortality and clinical signs were monitored for at least 6 hours after administration and then daily. All animals were weighed before treatment (day 1) and on days 2, 4, 6, 8, 11, 13, and 15. At the end of the observation period, all surviving rats were sacrificed and subjected to a detailed necropsy.

Results

No mortality occurred during the course of this study.

No clinical signs of toxicity were observed.

The body weight development was inconspicuous throughout the study.

The gross pathological examination revealed no organ alterations.

Conclusion

The test item has no acute toxic potential under the conditions of the present study, and the LD50 value is higher than 2000 mg/kg bw after single oral administration in female rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Guideline study under GLP conditions

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:
August 31, 2016 - March 28, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
February 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
Council Regulation (EC) No. 440/2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006 of the European Parliament and the council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item preparation was prepared directly before administration. Appropriate amounts of the test item were moistened sufficiently with the vehicle and grounded in a mortar using a pestle. The test item preparation was administered within less than 1 hour after preparation. The stability of the test item in the vehicle was not investigated.
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Sulzfeld, Germany
- Age at study initiation: 9 weeks (males), 12 weeks (females)
- Weight at study initiation: The mean initial body weight at the start of study was 226 g (range from 199 to 265 g).
- Fasting period before study: no
- Housing: separately in type III Makrolon cages with a shelter, placed on mobile racks
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.4 – 23.1°C
- Humidity (%): 40.9 – 59.7%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12 h

IN-LIFE DATES: From: day 1 To: day 15
Type of coverage:
semiocclusive
Vehicle:
paraffin oil
Details on dermal exposure:
TEST SITE
- Area of exposure: 39 cm2
- % coverage: approx. 10%
- Type of wrap if used: self-adhesive fabric (Fixomull stretch, Beiersdorf)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50, 200, 1000 and 2000 mg/kg bw
- For solids, paste formed: yes

VEHICLE
Designation: Art. 1.07174
Synonym: Liquid paraffin
Supplier: Merck KGaA, Germany
Batch: K45409474
Released until: April 30, 2019


Duration of exposure:
24 h
Doses:
50, 200, 1000 and 2000 mg/kg bw
No. of animals per sex per dose:
50 mg/kg bw: 5 (f)
200 mg/kg bw: 5 (f)
1000 mg/kg bw: 5 (f)
2000 mg/kg bw: 5 (m) / 5 (f)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 2, 4, 6, 8, 11, 13, and 15 of the experimental part.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Statistics:
Standard statistical methods have been applied for data processing.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the course of this study.
Clinical signs:
No clinical signs of toxicity were observed.
Body weight:
One female rat treated with 2000 mg/kg showed a decrease of the body weight during the observation period. The body weight development of all other rats was inconspicuous throughout the study.
Gross pathology:
Gross pathological examination revealed no organ alterations in all animals.
Interpretation of results:
GHS criteria not met
Conclusions:
The test item has no acute dermal toxic potential under the conditions of the present study, and the LD50 value exceeds 2000 mg/kg body weight after single dermal administration in rats.
Executive summary:

Objective

The objective of the present study was to identify potential toxic effects of the test item, after single dermal administration to rats in a stepwise procedure.

Study Design

Groups of 5 female rats each were dermally treated with the test item for 24 hours at dose levels of 50, 200, 1000 and 2000 mg/kg bw. The study was started with the lowest dose level of 50 mg/kg bw. As no mortalities were observed up to the highest dose, a further group of 5 male rats was exposed at a dose level of 2000 mg/kg bw.

Mortality and clinical signs were monitored for at least 6 hours after administration and then daily. All animals were weighed before treatment (day 1) and on days 2, 4, 6, 8, 11, 13, and 15. At the end of the observation period, all surviving rats were sacrificed and subjected to a detailed necropsy.

Results

No mortality occurred during the course of this study.

No clinical signs of toxicity were observed.

One female rat treated with 2000 mg/kg bw showed a decrease of the body weight during the observation period. The body weight development of all other rats was inconspicuous throughout the study.

Gross pathological examination revealed no organ alterations in all animals.

Conclusion

The test item has no acute dermal toxic potential under the conditions of the present study, and the LD50 value exceeds 2000 mg/kg body weight after single dermal administration in rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Guideline study under GLP conditions

Additional information

Justification for classification or non-classification

Based on the provided information the test item is not classified for acute toxicity after oral and dermal exposure according to the EU Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.