Registration Dossier

Administrative data

Description of key information

LD50 (oral, rat) > 2000 mg/kg

LC50 (inhaled, dust, rat) > 4.19 mg/L

LD50 (dermal, rat) > 2000 mg/kg

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:
13 October 2015 - 28 October 2015
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)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
- batch No.of test material: 141016


- Storage condition of test material: Room temperature 15-25°C, below 70 RH%


Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 196-206g

- Housing: 3 animals / cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5-6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4 – 24.8°C
- Humidity (%): 30 - 68%
- Air changes (per hr): 15 – 20 air exchanges/hour
- Photoperiod (hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Route of administration:
oral: gavage
Vehicle:
methylcellulose
Details on oral exposure:
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose.
Doses:
2000 mg/kg bw of B331
No. of animals per sex per dose:
6 females (2 groups of 3 animals)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter
- Necropsy of survivors performed: yes
- Other examinations performed: Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
B331 did not cause mortality at a dose level of 2000 mg/kg bw.
Clinical signs:
Treatment with B331 caused hunched back in all the experimental animals at the dose level of 2000 mg/kg bw. All animals were symptom free from 3 hours after treatment (Day 0). However coloured faeces (blackish) were observed for all the experimental animals on Day 1 (ascribed to the presence of test item in faeces).
Body weight:
There were no effects on body weights or body weight gains that could be attributed to treatment with of B331.
Gross pathology:
There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the test item B331 was found to be above 2000 mg/kg bw in female CRL:WI rats.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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:
17 March 2016 - 07 July 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Sighting study 11 weeks; main study 8 weeks
- Weight at study initiation: signthing: 402 g (male), 244 g (female); main study
- Fasting period before study: Not recorded
- Housing: Polycarbonate solid floor cages with stainless steel mesh. 5 animals per cage, separated by sex
- Diet: Unrestricted supply
- Water: Unrestricted supply
- Acclimation period: Animals were acclimated to laboratory conditions for 27 days (sighting study) or 14 days (main study) prior to involvement in the study. Animals were also acclimatised to the test apparatus (restrain procedures) for a short period prior to testing in order to lessen the stress during exposure.

ENVIRONMENTAL CONDITIONS
- Temperature: 19.0 - 25.9°C
- Humidity: 34 - 74% RH
- Air changes: At least 15 air changes per hour
- Photoperiod: 12 hours of continuous artificial light in each 24-hour period (from 6.00 a.m. to 6.00 p.m.)

IN-LIFE DATES: From: 12 May 2016 (receipt of first animals) To: 07 July 2016 (last necropsy occasion)
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
clean air
Mass median aerodynamic diameter (MMAD):
>= 1 - <= 4 µm
Geometric standard deviation (GSD):
>= 1.5 - <= 3
Remark on MMAD/GSD:
Measurements of aerodynamic particle size were performed from the animal’s breathing zone using a cascade impactor.
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
The test item was aerosolized using a dust generator according to Wright located at the top of the exposure chamber. In this device a fixed blade scrapes continuously the surface of the test item compacted by hydraulic press in a rotating reservoir. Dispersion was carried out by a high velocity air flow inside the outlet nozzle.
Dried compressed air was supplied by means of an oil-free compressor and passed through a suitable filter system prior to introduction to the nebuliser.

TEST ATMOSPHERE
- Brief description of analytical method used: Dust concentration was measured gravimetrically. Samples were collected at 10-20 minute intervals during the 4-hour exposure period
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- MMAD (Mass median aerodynamic diameter): 3.71 µm (sighting study); 3.92 µm (main study)
Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
4 h
Concentrations:
Achieved concentration: 4.23 mg/L sighting study, 4.19 mg/L main study
No. of animals per sex per dose:
5 main study, 1 (sighting study)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were checked hourly during exposure, 1 hour after exposure and twice daily (early and late in the working day) during the 14-day observation period for morbidity and/or mortality.
- Necropsy of survivors performed: yes
- Other examinations performed:
Individual body weights were recorded prior to treatment on the day of exposure (Day 0) and on Days 1, 3, 7 and 14.

All animals were observed for clinical signs at hourly intervals during exposure whilst the animals were still restrained. Following exposure, clinical observations were performed twice on the day of exposure (following removal from the restrainer and approximately one hour after completion of the exposure) and subsequently once daily for 14 days. Observations included changes in the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, somato-motor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4.19 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortality occurred during the study.
Clinical signs:
other: Fur staining by the test item occurred anywhere in study animals were considered to be related to the restraint and exposure procedures but not to be toxicologically significant. Laboured respiration (slight) was recorded in both animals of the Group 0.1
Body weight:
Slight body weight losses, noted in any study animals of both Group 0.1 and Group 1 on Day 0-1, were considered to be related to the restraint and exposure procedures.
From Day 3, positive body weight gain was recorded in all rats of the study.
Gross pathology:
A single four hours nose-only exposure of B331 to Crl:WI rats followed by a 14-day observation period produced test item-related gross changes. Red discoloration of the non-collapsed lungs and blue/dark blue discoloration of the lung-associated lymph nodes were noted at the concentration of 4.23 mg/L during sighting exposure. In main study, blue diffuse discoloration of the non-collapsed lungs and blue diffuse discoloration of the tails were recorded at the dose level of 4.19 mg/L. The blue discoloration was related to the administration of the test item and was attributed to its appearance (blue powder)
Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, no mortality occurred in Group 1 (main study) when exposed to a test atmosphere concentration of 4.19 mg/L as a maximum feasible concentration for 4 hours. The acute inhalation median lethal concentration (LC50) of B331 in Crl:WI rats was therefore considered to be above 4.19 mg/L.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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:
30 June 2016 - 20 July 2016
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:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 7-8 weeks old
- Weight at study initiation: Between 218 g and 244 g
- Housing: Individual caging
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0 – 24.1°C
- Humidity (%): 30 – 70 %
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 30 June 2016 To: 20 July 2016
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment. The test item was applied as a single dose, moistened with water to the shaved skin and remained in contact with the skin for the 24-hour exposure period. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours. At the end of the exposure period, the treated area of skin with the test item was washed with water of body temperature.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
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: On the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, skin irritation, body weight,
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Test item did not cause mortality at the dose level of 2000 mg/kg bw.
Clinical signs:
Systemic Clinical Signs: There were no systemic clinical signs noted in any animal throughout the study.
Local Dermal Signs: The test item coloured the surface of the skin to bluish for 3-6 days. No adverse local dermal signs were observed after treatment with the test item during the 14 days observation period.
Body weight:
There were no treatment related effects on body weight or body weight gain during the observation period.
Gross pathology:
There was no evidence of any gross changes at a dose level of 2000 mg/kg bw.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item B331 was found to be greater than 2000 mg/kg body weight in male and female Crl:WI rats.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Justification for classification or non-classification

Three relevant acute toxicity studies are available to assess the acute toxicity of B331. No mortality was seen in an acute toxicity study in rats dosed orally at 2000 mg/kg; no mortality was observed in rats dosed by inhalation at 4.19 mg/L; no mortality was observed in rats receiving a dermal dose of 2000 mg/kg. In accordance with Regulation (EC) 1272/2008 including amendments to May 2017, Annex I Part 3.1, B331 does not meet the criteria for classification as acutely toxic.

No specific studies are available to address specific target organ toxicity following a single exposure (STOT-SE) however it is noted that in each of the available acute toxicity studies no significant gross pathological signs or clinical observations were recorded which would indicate significant sub-lethal toxicity and no target organ was apparent.