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Administrative data

Description of key information

Oral:
The oral LD50 of the test substance in rat was established to be in the range from 300 to 2000 mg/kg bw.
Inhalation:
The inhalation LD50 of the test substance in rat was found to be 5.31 mg/L aerosol.
Dermal:
The dermal LD50 of the test substance in rats was determined to be greater than 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:
2015-07-29 to 2015-09-14
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)
Version / remarks:
2001
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: 8-9 weeks
- Weight at study initiation: 170.0-211.1 g
- Fasting period before study: overnight
- Housing: single, Stainless wire mesh cage, 260W*350D*210H (mm)
- Diet: ad libitum, Pelleted rodent chow, Harlan Laboratories Inc. U.S.A.
- Water: ad libitum, tap water
- Acclimation period: in quarantine room 7 days, in animal room 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5-23.2
- Humidity (%): 46.3-61.3
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 60 and 400 mg/mL
- Amount of vehicle: 5 mL/kg bw
- Lot/batch no.: MKBS6944V

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: 300 mg/kg bw was selected as starting dose
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
6 for 300 mg/kg bw, 3 for 2000 mg/kg bw, all female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observation: 30 min, 1, 2, 4 and 6 h, and once daily after administration; weighing: before, on day 1, 3, 7 and 14 after administration
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology
Statistics:
Statistical analysis was not performed.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Sex:
female
Dose descriptor:
LD0
Effect level:
300 mg/kg bw
Based on:
test mat.
Sex:
female
Dose descriptor:
LD100
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Mortality:
2000 mg/kg bw: 2 animals dead on day 2, 1 on day 3
300 mg/kg bw: all animals survived
Clinical signs:
300 mg/kg bw: Mucous stool in 4 animals 2 h after application and in 2 animals 4 h after application. Soiled perineal region and decreased fecal volume were observed on day 1. From day 2 onward no clinical signs were observed.
2000 mg/kg bw: Abnormal gait, lacrimation, salvation, decreased fecal volume, tremor, tonic convulsion, hypersensitivityy, loss of locomotor activity, lying on side and hypothermia were observed starting 4-6 h after treatment and worsening on day 1 and 2 after application.
Body weight:
300 mg/kg bw: decrease ob body weight or supressed body weight gain were observed in 3 animals on day 1. Normal body weight gain was obserevd from day 3 onwards.
Gross pathology:
300 mg/kg bw: no abnormalities were detected.
2000 mg/kg bw: 2 animals had mottled yellowish-white or pale livers. Histopathology of the livers showed minimal or moderate hepatocellular vacuolation. Those hepatocytes had foamy cytoplasm and were distributed diffusely in hepatic lobes.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral LD50 of the test substance was established to be in the range from 300 to 2000 mg/kg bw in rats.
Executive summary:

The acute oral toxicity of the test substance was investigated with the acute toxic class method according to OECD Guideline 423 in rats. Experiments were conducted using GLP. The test substance was administered via gavage at 300 and 2000 mg/kg bw and the animals were observed for 14 days after treatment. All animals treated with 300 mg/kg bw showed mucous stool after administration. They recovered and no clinical signs were observed from day 2 onwards. Animals treated with 2000 mg/kg bw showed various clinical signs, e.g. loss of motor activity and hypothermia, and died on day 2 or 3 after application. The acute oral LD50 for rats was established to be in the range from 300 to 2000 mg/kg bw. Therefore the test substance was classified in category 4. H302: Harmful if swallowed

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
300 mg/kg bw
Quality of whole database:
The available study is sufficient for assessment.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-03-16 to 2015-03-31
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
2009
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: 10-11 weeks
- Weight at study initiation: Males: 253.4 to 298.6 g, Females: 191.1 to 218.6 g
- Fasting period: during study
- Housing: individually, standard polysulfone cages (Size: approximately L 425 x B 266 x H 185 mm),
- Diet: ad libitum, Teklad Certified (2014C) Global 14 % Protein Rodent Maintenance Diet -Pellet (Certified) manufactured by Harlan Laboratories B.V. AN Venray, The Netherlands
- Water: ad libitum, deep bore-well water passedc through charcoal filter an exposed to UV rays
- Acclimation period: 6-7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-24
- Humidity (%): 65-67
- Air changes (per hr): 12-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
clean air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: nose only exposure chamber
- Exposure chamber volume: 53 L
- Method of holding animals in test chamber: restrainers, polycarbonate tubes having facility to trap the faeces for individual exposure tubes, sizes males: length 19.0 cm, diameter 6.0 cm, females: length 15.0 cm, diameter 6.0 cm
- Source and rate of air: Dehumidified and filtered air from air compressor, 20 L/min
- Method of conditioning air: injection (0.2, 0.4 and 0.8 mL/min) and atomizing (Atomizer pressure 1.4 kg/cm2)
- System of generating particulates/aerosols: A glass atomizer was used. Manufactured by: Sai Scientifics Bengaluru Specification: Injection capacity of 1.6 mL/minute.
- Method of particle size determination: Instrument: GALAI CIS-50 particle size analyzer, Manufactured by: Galai Pvt. Ltd., Israel, Principle of measurement: Laser based 'Time-of-Transition Theory', Unit of measurement: Aerosol particle size in µm
- Treatment of exhaust air: collected in aerosol exhaust chamber and via filters to the exhaust system
- Temperature, humidity in air chamber: inner chamber: 22.9-25.3 °C, 73.4-77.5 % relative humidity, outer chamber: 19.9-26.8 °C, 64.9-78.3 % relative humidity

TEST ATMOSPHERE
- Brief description of analytical method used: during exposure areosol particle size, oxygen content in chamber air,
- Samples taken from breathing zone: yes
- Mean aerosol particle size: 0.2 mL/min: 1.79 ± 0.97 µm, 0.4 mL/min: 1.83 ± 0.97 µm, 0.8 mL/min: 1.73 ± 1.00 µm
- Geometric standard deviation (GSD): 0.2 mL/min: 1.99, 0.4 mL/min: 1.99, 0.8 mL/min: 2.19

A preliminary test with 6 (3M/3F) animals was performed to select the concentration for the main test. The concentration tested was 0.4 mL/min.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
test item injection rate was 0.2, 0.4 and 0.8 mL/min
average concentration: 0.2 mL/min = 3.31 mg/L, 0.4 mL/min = 9.23 mg/L, 0.8 mL/min = 10 mg/L
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: observation: during exposure 1/h, directly after and 1 h after exposure, once daily from day 2-15, body weight: during acclimatisation, pre-exposure (day 1) and on day 2, 4, 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Statistics:
No statistical analysis was performed.
Preliminary study:
A preliminary test with 6 (3m/3f) animals was performed to select the concentration for the main test. The concentration tested was 0.4 mL/min. Animals showed Clear nasal discharge, slight salivation and moderate ataxia on day 1. Hypoactivity, arched back, slight /moderate ataxia and convulsions were observed. One female rat died on day 3.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
5.31 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
0.2 mL/min: no mortality during exposure, 3 male and 1 female died on day 2
0.4 mL/min: no mortality during exposure, 3 male and 1 female died on day 2, 2 females died on day 3
0.8 mL/min: no mortality during exposure, 3 male and 3 females died on day 2, 2 females died on day 3, 1 male died on day 4
Clinical signs:
other: 0.2 mL/min: clear nasal discharge, hypoactivity, slight tremors, slight salivation, slight/severe ataxia and slight piloerection; From day 4/5/6/7 onwards no clinical signs were observed. 0.4 mL/min: clear nasal discharge, hypoactivity, slight tremors, sl
Body weight:
0.2 mL/min: day 2 all decrease, day 4 decrease (one increase), day 8 decrease (three increase), day 15 all increase
0.4 mL/min: days 2, 4 and 8 all decrease, day 15 all increase
0.8 mL/min: 1 surviving rat: decrease on day 2, 4 and 8, increase on day 15
All animals that died during the observation time had a decreased body weight compared to the weight before exposure.
Gross pathology:
0.2 mL/min: In one preterminally dead male lung congestion was observed. All other animals showed no abnormalities.
0.4 mL/min: In two preterminally dead male and 1 female lung congestion was observed. All other animals showed no abnormalities.
0.8 mL/min: In three preterminally dead male and three female lung congestion was observed. All other animals showed no abnormalities.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute inhalation LD50 of the test substances aerosol was established to be 5.31 mg/L in rats.
Executive summary:

The acute inhalation toxicity of the test substance was investigated according to OECD Guideline 403 in rats. Experiments were conducted using GLP. In each case 10 animals were exposed for 4 h to an aerosol of test substance at concentrations of 3.31 mg/L (0.2 mL/min), 9.23 mg/L (0.4 mL/min) and 10 mg/L (0.8 mL/min). The animals were observed for 14 days after treatment. Animals exposed to 3.31 mg/L showed clear nasal discharge, hypoactivity, slight tremors, slight salivation, slight/severe ataxia and slight piloerection. 4 animals died on the day after exposure while the surviving animals recovered and no clinical signs were observed from day 7 onwards. Animals exposed to 9.23 and 10 mg/L showed clear nasal discharge, hypoactivity, slight tremors, slight/moderate salivation, slight/severe ataxia and slight piloerection. After exposure to 9.23 mg/L 6 animals died within the first 2 days and 9 animals died within the first 3 days after exposure to 10 mg/L. The surviving animals recovered and showed no clinical signs after day 8/9 after exposure. In all animals a decrease in body weight was detected after exposure. The surviving animals showed an increase in body weight 14 days after exposure. The acute inhalation LD50 for rats was established to be 5.31 mg/L aerosol. Therefore the test substance is not classified. 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
5 310 mg/m³
Quality of whole database:
The available study is sufficient for assessment.

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:
1990-06-14 to 1990-06-28
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1981
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: appr. 10-14 weeks
- Weight at study initiation: 218-243 g
- Housing: individually in solid-floor polypropylene cages furnished with soft wood shavings.
- Diet: ad libitum, Rat and Mouse Expanded Diet No. 1, Special Diet Services Limited, Witham, Essex, U.K.
- Water: ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 60-77
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: back and flanks
- % coverage: appr. 10 %
- Type of wrap if used: A piece of surgical gauze measuring 7 cm x 4 cm was placed over the treatment area and semioccluded with a piece of self-adhesive bandage (HYPERTIE). The bandage was further secured with a piece of BLENDERM wrapped around each end.

REMOVAL OF TEST SUBSTANCE
- Washing: after removal of the bandage the treated skin and surrounding hair was wiped with cotton wool moistened with distilled water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 2.07 mL/kg bw, dose level: 2000 mg/kg bw

Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical observations were performed 1, 2, 3 and 4 hours after dosing and daily afterwards, body weights were determined weekly
- Necropsy of survivors performed: yes
Statistics:
Not required.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occured.
Clinical signs:
No abnormalities were detected.
Body weight:
No toxicologically significant effects an bodyweight were noted during the study.
Gross pathology:
No abnormalities were noted at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test material in the rat was found to be greater than 2000 mg/kg bw.
Executive summary:

A study was performed to assess the acute dermal toxicity of the test material in the Sprague-Dawley strain rat. The study was performed according to OECD Guideline 402. A group of ten animals (five males and five females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. No signs of systemic toxicity or skin irritation were noted during the study and no deaths occurred. No toxicologically significant effects on bodyweight were noted during the study and no abnormalities were noted at necropsy of animals killed at the end of the study. Therefore, the acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
The available study is sufficient for assessment.

Additional information

Oral:

Key study

The acute oral toxicity of the test substance was investigated with the acute toxic class method according to OECD Guideline 423 in rats. The test substance was administered via gavage at 300 and 2000 mg/kg bw and the animals were observed for 14 days after treatment. All animals treated with 300 mg/kg bw showed mucous stool after administration. They recovered and no clinical signs were observed from day 2 onwards. Animals treated with 2000 mg/kg bw showed various clinical signs, e.g. loss of motor activity and hypothermia, and died on day 2 or 3 after application. The acute oral LD50 for rats was established to be in the range of 300 to 2000 mg/kg bw. Therefore the test substance was classified in category 4.

Supporting studies

A study (according to OECD Guideline 401) was performed to determine the acute oral LD50 of the test material in rats. A group of ten fasted animals was given a single oral dose of test material preparation at a dose level of 200 mg/kg bodyweight. No deaths occurred. Signs of toxicity noted were confined to hunched posture and pilo-erection during the day of dosing. All animals appeared normal one day after treatment and no further abnormalities were detected. The oral LD50 of the test material in rats was found to be greater than 200 mg/kg bw. As no classification and labelling can be derived from this result, the study was considered as supporting information.

Conclusion

The OECD 423 guideline conform study conducted in rats was selected as key study for acute oral toxicity. In this study a LD50 in the range from 300 to 2000 mg/kg bw was determined. The oral study according to OECD 401 in rats was selected as supporting study as the test method was not the preferred method to determine the acute oral LD50 and only one dose was administered which was below the 2000 mg/kg bw limit for discriminating dose tests. As the study determined a LD50 greater than 200 mg/kg bw the result is considered to support the key study but does not allow a further refinement of the LD50 range.

Taken together both studies show LD50 values in the same range and support the same classification. The most reliable study was chosen as key study for the reasons mentioned above.

 

Inhalation:

The acute inhalation toxicity of the test substance was investigated according to OECD Guideline 403 in rats. In each case 10 animals were exposed for 4 h to an aerosol of test substance at concentrations of 3.31 mg/L (0.2 mL/min), 9.23 mg/L (0.4 mL/min) and 10 mg/L (0.8 mL/min). The animals were observed for 14 days after treatment. All animals showed several clinical signs of toxicity. 4 animals died on the day after exposure while the surviving animals recovered and no clinical signs were observed from day 7 onwards After exposure to 9.23 mg/L 6 animals died within the first 2 days and 9 animals died within the first 3 days after exposure to 10 mg/L. The surviving animals recovered and showed no clinical signs after day 8/9 after exposure. In all animals a decrease in body weight was detected after exposure. The surviving animals showed an increase in body weight 14 days after exposure. The acute inhalation LD50 for rats was established to be 5.31 mg/L aerosol. Therefore the test substance is not classified. 

 

Dermal:

A study was performed to assess the acute dermal toxicity of the test material rats. The study was performed according to OECD Guideline 402. A group of ten animals was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bw. No signs of systemic toxicity or skin irritation were noted during the study and no deaths occurred. No toxicologically significant effects on bodyweight were noted during the study and no abnormalities were noted at necropsy of animals killed at the end of the study. Therefore, the acute dermal LD50 of the test material in rats was determined to be greater than 2000 mg/kg bw.

Justification for classification or non-classification

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

The available experimental test data is reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.

Based on available data for acute oral toxicity the substance is considered to be classified for acute oral toxicity into category 4 and labeled with H302 (Harmful if swallowed) according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in Regulation (EU) No 2016/918.

Based on available data on acute toxicity via inhalation and dermal route, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in Regulation (EU) No 2016/918.