Registration Dossier

Administrative data

Description of key information

Oral:
One study is available (Latour, 2014). The oral LD50 value in Wistar rats was established to exceed 2000 mg/kg bodyweight.
Inhalation:
One study is available (van Huygevoort, 2014). The inhalatory LC 50, 4 h value in Wistar rats was established to exceed 5 mg/L.
Dermal:
One study is available (Latour, 2014). The dermal LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.

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 01 to 21 May 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a method comparable with current guidelines and to GLP
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approx. 10 or 11 weeks old
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean
- Fasting period before study: Overnight prior to dosing
- Housing: Labeled Makrolon cages containing sterilized sawdust as bedding material and paper as cage-enrichment
- Diet (e.g. ad libitum): Pelleted rodent diet
- Water (e.g. ad libitum): Tap water
- Acclimation period: At least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): at least 10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

IN-LIFE DATES: From 01 to 21 May 2014
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Justification for choice of vehicle: The vehicle was selected based on trial formulations formed at WIL Research Europe and on test substance data supplied by the sponsor

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females.
The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines.
Doses:
2000 mg/kg
No. of animals per sex per dose:
3 females at a dose level of 2000 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: yes, at the end of observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy.
- Other examinations performed: no.
Statistics:
No statistical analysis was performed.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture and pioerection were noted for all animals between Day 1 and 12.
Lean appearance was noted for one animal between Days 7 and 9.
Body weight:
Most animals showed bodyweight loss between Days 1 and 8. All animals gained bodyweight between Days 8 and 15.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
No information provided.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 value of the test substance in Wistar rats was established to exceed 2000 mg/kg bodyweight.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
1 (reliable without restriction)

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:
From 05 to 19 Jun 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a method comparable with current guidelines and to GLP
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles Rivers Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 12 weeks old
- Weight at study initiation: body weight did not exceed +/- 20% of the sex mean
- Fasting period before study: no
- Housing: labelled Makrolon cages
- Diet (e.g. ad libitum): pelleted rodent diet
- Water (e.g. ad libitum): tap water
- Acclimation period: At least 5 days before start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 40 - 70
- Air changes (per hr): at least 10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12-hour light / 12-hour dark cycle

IN-LIFE DATES: From 05 to 19 June 2014
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The design of the exposure chamber is based on the flow past nose-only inhalation chamber. The chamber consisted of three animal sections with eight animal ports each. Each animal port had its own atmosphere inlet and exhuast outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and numver of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
- System of generating particulates/aerosols: The test substance was fed to a stream of dried pressurized air by means of a accurate dust feeder and a micronizing jet mill. The aerosol was passed through two cyclones, in order to allow larger particles to settle, and subsequently entered the exposure chamber.
- Method of particle size determination: The paricle size distribution was characterized twice during the each exposure period. The samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters and a fiber glass back-up filter. Amounts of test substance collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter and the Geometric Standard Deviation were determined.
- Temperature, humidity, pressure in air chamber: The temperature of the atmosphere during the exposure was between 19.6 and 20.2 °C and relative humidity was between 36 and 38%.

TEST ATMOSPHERE
- Brief description of analytical method used: The nominal concentration was calculated by dividing the amount of test substance used by the volume of pressurized air entering the exposure chamber used for exposure of the animals. Due to the small volumn of the exposure chamber the equilibrium time was negligible. The volumn of air was calculated from the averaged air flow and the exposure time. The acutual concentration was determined twenty two times during the exposure period. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. Samples were drawn through a glass fiber filter. The collected amount of the test substance in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter. Subsequently the time-weighted mean concentration with the standard deviation was calculated.

VEHICLE
- Composition of vehicle (if applicable): dried pressurized air
- Concentration of test material in vehicle (if applicable): 5 mg/L
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
5 mg/L
No. of animals per sex per dose:
5 animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality: twice daily.
Clinical signs: During exposure: 3 times for mortality, behavioural signs of distress and effects on respiration. After exposure: on day 1, one and three hours after exposure and once daily thereafter until day 15.
Body weight: days 1 (pre-administration), 2, 4, 8 and 15.
- Necropsy of survivors performed: yes, all animals were sacrificed at the end of the observation period by an intraperitoneal injection with Euthasol. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded. Paticular attention was given to any changes in the respiratory tract.
- Other examinations performed: no.
Statistics:
No statistical analysis was performed.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortality occurred.
Clinical signs:
During exposure, no clinical signs were seen. After exposure, all animals showed hunched posture between days 1 and 5.
Body weight:
Reduced body weight gain and body weight loss was noted for the animals up to Day 8. All animals regained weight during the second week, except for one female that did not gain weight.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The inhalatory LC 50, 4 h value of the test substance in Wistar rats was established to exceed 5 mg/L.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
5 000 mg/m³
Quality of whole database:
1 (reliable without restriction)

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 30 Apr to 14 May 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a method comparable with current guidelines and to GLP
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approx. 10 weeks old
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean
- Fasting period before study:
- Housing: labeled Makrolon cages containing sterilized sawdust as bedding material and paper as cage-enrichment
- Diet (e.g. ad libitum): pelleted rodent diet
- Water (e.g. ad libitum): tap water
- Acclimation period: at least 5 days before start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): at least 10 air changes/hour
- Photoperiod (hrs dark / hrs light): a 12-hour light/12-hour dark cycle

IN-LIFE DATES: From 30 Apr to 14 May 2014
Type of coverage:
occlusive
Vehicle:
propylene glycol
Details on dermal exposure:
TEST SITE
- Area of exposure: 25 cm2 for males and 18 cm2 for females
- % coverage: 10% of the total body surface
- Type of wrap if used: a dressing, consisting of a surgical gauze patch, successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): tap water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg (10 mL/kg) body weight
- Concentration (if solution): 2000 mg/kg (10 mL/kg) body weight
- Constant volume or concentration used: yes
Duration of exposure:
24 hours
Doses:
2000 mg/kg (10 mL/kg) body weight
No. of animals per sex per dose:
5 animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
Body weights: Days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes, at the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
None stated
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Scabs or scales were seen in the treated skin-area of all female animals between Days 5 and 15.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
No information provided.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 value of the test substance in Wistar rats was established to exceed 2000 mg/kg body weight.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
1 (reliable without restriction)

Additional information

Oral: One study is available which was conducted according to OECD Guideline 423 under GLP (Latour, 2014). The oral LD50 value in Wistar rats was established to exceed 2000 mg/kg bodyweight.

Inhalation: One study is available which was conducted according to OECD Guideline 403 under GLP (van Huygevoort, 2014). The inhalatory LC 50, 4 h value in Wistar rats was established to exceed 5 mg/L.

Dermal: One study is available which was conducted according to OECD Guideline 402 under GLP (Latour, 2014). The dermal LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.


Justification for selection of acute toxicity – oral endpoint
Study run to a method comparable with current guidelines and to GLP

Justification for selection of acute toxicity – inhalation endpoint
Study run to a method comparable with current guidelines and to GLP

Justification for selection of acute toxicity – dermal endpoint
Study run to a method comparable with current guidelines and to GLP

Justification for classification or non-classification

Oral: rat LD50 > 2000 mg/kg bw (actual value > 2000 mg/kg bw);

Dermal: rat LD50 > 2000 mg/kg bw (actual value > 2000 mg/kg bw);

Inhalation: dust LC50 > 5 mg/L (actual value > 5 mg/L).

Therefore in accordance with Regulation (EC) No. 1272/2008 (as amended by Regulation (EC) No. 286/2011) Table 3.1.1, this substance should not be classified for acute toxicity endpoints.