Registration Dossier

Administrative data

Description of key information

The acute oral, inhalation and dermal toxicity studies indicate that the substance is of low  toxicity if swallowed (rat LD0 > or = 2000 mg/kg bw), applied onto the skin (rat LD0 > or = 2000 mg/kg bw) or inhaled (rat LC0> or = 5110 mg/m3)

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:
2010
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)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: 8 weeks old
- Weight at study initiation: From 180 to 220g (± 20%)
- Fasting period before study: the animals were fasted during the night before treatment but had free access to water. Fodd was given back approximately 4 hours after administration of the test item.
- Housing: in polycarbonate cages with a stainless steel lid (43 x 22 x 20 cm).
- Diet (e.g. ad libitum): ad libitum (SSNIFF R/M-H pellet diet)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: no data
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
VEHICLE
The solubility assay first started at the concentration of 200 mg/mL, and the first choice vehicle was drinking water treated by reverse osmosis.
Since a heterogeneous suspension at 200 mg/mL was obtained with drinking water treated by reverse osmosis, the test substance was tested with a 0.5% methylcellulose aqueous solution.
A homogenous suspension was obtained at the concentration of 200 mg/mL in a 0.5% methylcellulose aqueous solution.

DOSAGE PREPARATION (if unusual): The test substance was administered as a homogeneous suspension in the vehicle. The test substance was ground to a fine powder, using a mortar and pestle, and then mixed with the required quantity of vehicle.
Dose formulations preparations were prepared extemporaneously on the day of each administration.
The dose formulations were stored at room temperature and delivered to the study room in brown flasks.

CLASS METHOD (if applicable):
- Rationale for the selection of the starting dose:
Since no relevant toxicity data were available for the estimation of a lethal dose-level and any existing data were taken into account by the Sponsor, the starting dose level was 300 mg/kg for ethical reasons.
After treatment at the starting dose-level, the next higher dose-level of 2,000 mg/kg was tested.

ADMINISTRATION:
The dose formulations were administered once by gavage, using a plastic syringe fitted with a plastic gavage tube. The quantity of dose formulation administered to each animal was adjusted according to the bodyweight recorded on the day of the dose administration.
A constant dosage-volume of 10 mL/kg was used.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
300 mg/kg bw: 3 females
2000 mg/kg bw: 3+3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Clinical observations: At least once during the first 30 minutes, periodically during the first 4 hours, then once a day, at approximately the same time.
- Morbidity and mortality: Frequently during the hours following administration, then at least once a day until the end of the observation period, including weekends and public holidays.
- Bodyweight: Just before treatment on Day 1; then on Days 8 and 15.
- Necropsy of survivors performed: Yes (macroscopic). The main organs included the digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities. All gross observations were recorded individually for each animal.
Statistics:
None
Key result
Sex:
female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occured in any animals at any tested dose.
Clinical signs:
No clinical signs were observed.
Body weight:
When compared to laboratory historical control data, a lower body weight gain was noted between day 1 and day 8 in 1/6 females given 2000 mg/kg bw (30 g vs 41 +/- 9g in control data base). The body weight gain returned to normal thereafter. The body weight gain of the other animals given 300 or 2000 mg/kg bw was not affected by test-item treatment.
Gross pathology:
At necrospsy, in one female given 2000 mg/kg bw, both horns of uterus were dilated and had translucent content. No apparent abnormalities were observed in the other females given 300 or 2000 mg/kg bw.
Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD0 of the test substance was equal or higher than 2000 mg/kg bw.
Executive summary:

In an acute oral toxicity study, performed according to the OECD 423 test guidelines and in compliance with GLP, groups of fasted Sprague-Dawley female rats were administered a single oral dose of the test substance in 0.5 % methylcellulose by gavage. The animals were observed for mortality, clinical signs and body weight for 14 days and then necropsied for macroscopic observations.

Three animals were first given an oral dose of 300 mg/kg bw. As no mortality nor clinical signs were observed, a higher dose was tested. Therefore two independent and successive groups of 3 female rats each were given the maximum dose of 2000 mg/kg bw. Again, no mortality nor clinical signs were observed at 2000 mg/kg bw. A lower body weight gain was noted in 1/6 tested females between day 1 and 8 but the body gain returned to normal thereafter. At necropsy, one female treated with 2000 mg/kg bw, presented a dilatation of both horns of uterus accompagnied of a translucent content. Since no apparent abnormalities were observed in the other animals at the both given doses, these effects were considered not significant.

The acute oral LD0 was found equal or greater than 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Study complete and sufficient to fulfill the endpoint requirements.

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 2010- 04-08 to 2010-07-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Remarks:
The age of the preliminary investigation animals may have been 6 weeks, and not 7 to 8 weeks as per protocol.
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: from Charles River (UK) Ltd
- Age at study initiation: 7-8 weeks
- Weight at study initiation: in the range of 308 to 341 g for males and 220 to 274 g for females.
- Fasting period before study: no data
- Housing: the animals were housed inside a restricted entry rodent facility, one animal per cage during the preliminary investigation and five of one sex per cage during the main study.
The cages were made of a polycarbonate body with a stainless steel mesh lid. Wood shavings were used as bedding and were sterilised by autoclaving and changed at appropriate intervals each week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): within the range of 19 to 23°C
- Humidity (%): within the range of 40 to 70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): a cycle of 12 hours continuous light and 12 hours continuous dark per 24 hours.

IN-LIFE DATES: no data
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
3.4 µm
Geometric standard deviation (GSD):
2.72
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: flow through nose only chamber. This system was an aluminium alloy construction comprising a base unit, one animal exposure section and a top section.
- Method of holding animals in test chamber: Rats were held in polycarbonate tubes with their snouts protruding from the end of the tubes into the exposure chamber.
- Source and rate of air: From in-house compressed air system(breathing quality), Generator flow: 25 L/minute
- System of generating particulates/aerosols: Turn Table Dust Feed mechanism designed to produce and maintain atmospheres of dust by suspending material in a stream of dry air. Test article flowed from a hopper, assisted by agitation, into a concentric groove in the turntable. As the turntable rotated, the dust passed under the uplift tube which draws up the material using the negative pressure supplied using the venturi principle. The material is carried in the air stream and into the chamber.The concentration of dust in the air may be altered by modifying the speed of rotation of the turntable plate. The Turn Table Dust Feed mechanism was attached to an aerosol conditioning chamber for the delivery of the powder and the aerosol passed through it into the top of the exposure chamber.
- Temperature, humidity, pressure in air chamber:
Temperature was measured from the breathing zone of the animals via an unused exposure port using an electronic thermometer. The mean chamber temperatures were 20.5 and 20.1°C for the control and dosed groups respectively.
Chamber Relative Humidity was measured using an electronic hygrometer inserted into the breathing zone of the animal via an unused exposure port. The mean chamber relative humidity were 21.4 and 21.2% for the control and dosed groups respectively.
Pressure in air chamber was not reported.
- Air flow rate: Air flow from Turn Table Dust Feed mechanism was 36 L/min
- Method of particle size determination: Particle size analysis of generated atmospheres was performed using a 8-stage cascade impactor (Marple 298). Samples were collected twice during exposure. The collection substrates and the backup filter were weighed before and after sampling and the weight of test item, collected at each stage, was calculated by this difference. The total amount collected for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than < 0.42, 0.76, 1.27, 2.86, 4.9, 8.0, 12.08 and 17.39 µm (aerodynamic diameter) was calculated. From this data, the Mass Median Aerodynamic Diameter (MMAD), and Geometric Standard Deviation (GSD) were calculated.
- Treatment of exhaust air: Extract airflow was drawn at a flow rate of 40 L/minute.

TEST ATMOSPHERE
- Brief description of analytical method used: The actual concentration of the generated atmosphere was measured gravimetrically during the exposure by pulling a suitable, known volume of test atmosphere in the exposure chamber, through a glass fibre filters. Sampling was performed eight times during the exposure. Samples were collected from a vacant animal exposure port (animals breathing zone). The difference in the pre- and post-sampling weights, divided by the volume of atmosphere sampled, was equal to the actual achieved test atmosphere concentration.

- Samples taken from breathing zone: yes, samples were collected from a vacant animal exposure port.

VEHICLE (if applicable)
- none
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Table 7.2.2/1
Duration of exposure:
ca. 4 h
Concentrations:
Target concentration: 5.0 mg/L
Mean achieved atmosphere concentration: 5.11 mg/L ; Standard deviation: 0.76
No. of animals per sex per dose:
Five/sex/dose
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed intermittently for signs of reaction to the test substance during exposure and at least twice daily throughout the observation period. The weight of each animal was recorded on Days 1 (prior to dosing), 2, 3, 4, 8 and 15.
- Necropsy of survivors performed: yes, animals were killed by an intraperitoneal overdose of sodium pentobarbitone followed by exsanguination. All animals were subjected to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities.
- Other examinations performed: Clinical signs were recorded during exposure, immediately after exposure and then at 1-hour and 2-hours post-exposure. Throughout the study, all cages were checked at least twice daily, once in the morning and again towards the end of the normal working day, for dead or moribund animals. The lungs (including the larynx and trachea) were dissected free, weighed and the weights recorded.
Statistics:
None
Preliminary study:
A preliminary exposure, with 1 male and 1 female rat, was conducted at the target (Limit test) concentration of 5 mg/L for a period of 4-hours in order to assess the likely response of rats to the test substance. As a concentration of 5.63 mg/L was tolerated by the preliminary investigation animals, the main study test animals were also exposed to the target concentration of 5 mg/L for a period of 4-hours. Control animals received a single 4-hour exposure air only.
Key result
Sex:
male/female
Dose descriptor:
LC0
Effect level:
>= 5.11 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
There were no unscheduled deaths.
Clinical signs:
Immediately following exposure all test animals had reduced body temperature, whilst 7/10, 5/10 and 5/10 animals had irregular breathing, hunched posture and post dose salivation staining, respectively. In addition chin rubbing, unsteady gait and partially closed eyelids were recorded in 4/10, 3/10 and 2/10 animals, respectively. These signs generally persisted to 1 and 2-hours post-exposure, however with decreasing incidence, and were no longer present from Day 2 onwards.
Ungroomed coat and noisy breathing/râles were recorded at 2-hours post-exposure in 10/10 and 6/10 animals respectively, and persisted up to Days 3 and 5 respectively, however with decreasing incidence over time. In addition gasping and underactive behaviour were noted in 3/10 and 1/10 animals respectively at 2-hours post-exposure, whilst distended abdomen was recorded in one male and one female; these signs were not present at Day 2, with the exception of one male which had a distended abdomen and was gasping up to Day 2/3.
Piloerection, recorded in all test animals, was generally observed from 2-hours post-exposure, and persisted up to Day 4.
Body weight:
Bodyweight loss was recorded for all treated animals on Day 2 (7 to15% of pre-exposure Day 1 weights). On Day 3, one treated male had lost further weight (-19% of pre-exposure Day 1 weight), however all other treated animals had gained weight between Days 2 and 3. Subsequently, from Day 4 post-exposure all treated animals gained weight during the remainder of the observation period.
Gross pathology:
There were no macroscopic findings in the animals.
Other findings:
- Organ weights: Mean lung weights were slightly higher in treated animals (1.09X and 1.14X control, in males and females, respectively)

Table 7.2.2/1: Clinical signs in male animals

 

 

Number showing signs

Males

Signs

Time in hours

 

 

During exposure

IAE

1.0

2.0

Day 2

Day 3

Day 4

 

 

 

 

 

 

 

 

 

Control

Wet fur

0

2

0

0

0

0

0

 

 

 

 

 

 

 

 

 

Treated

Salivation

1

0

0

0

0

0

0

 

Wet fur

0

2

2

0

0

0

0

 

Chin rubbing

0

2

0

0

0

0

0

 

Post salivation staining

0

3

0

0

0

0

0

 

Reduced body temperature

0

5

0

0

0

0

0

 

Irregular breathing

0

2

0

1

0

0

0

 

Partially closed eyelids (both)

0

2

3

0

0

0

0

 

Unsteady gait

0

1

0

0

0

0

0

 

Piloerection

0

0

0

4

5

5

1

 

Ungroomed

0

0

0

5

1

1

0

 

Rales/noisy breathing

0

0

0

2

2

1

0

 

Gasping

0

0

0

1

1

1

0

 

Distended abdomen

0

0

0

1

1

0

0

 

Brown staining snout

0

0

0

0

1

1

0

IAE:    Immediately after exposure

Table 7.2.2/2: Clinical signs in female animals

 

 

Number showing signs

Females

Signs

Time in hours

 

 

During exposure

IAE

1.0

2.0

Day 2

Day 3

Day 4

Day 5

 

 

 

 

 

 

 

 

 

 

Control

Wet fur

0

3

0

0

0

0

0

0

 

 

 

 

 

 

 

 

 

 

Treated

Wet fur

0

3

2

1

0

0

0

0

 

Irregular breathing

5

5

5

1

0

0

0

0

 

Hunched posture

0

5

3

0

0

0

0

0

 

Chin rubbing

0

2

0

0

0

0

0

0

 

Reduced body temperature

0

5

2

0

0

0

0

0

 

Post salivation staining

0

2

3

0

0

0

0

0

 

Piloerection

0

1

2

3

4

5

1

0

 

Test substance staining

0

2

0

0

0

0

0

0

 

Gasping

0

1

0

2

0

0

0

0

 

Unsteady gait

0

2

2

0

0

0

0

0

 

Partially closed eyelids (both)

0

0

2

0

0

0

0

0

 

Ungroomed

0

0

0

5

2

2

0

0

 

Rales/noisy breathing

0

0

0

4

3

2

2

1

 

Brown staining snout

0

0

0

3

4

2

0

0

 

Underactive

0

0

0

1

0

0

0

0

 

Distended abdomen

0

0

0

1

0

0

0

0

 

 

 

 

 

 

 

 

 

 

IAE:    Immediately after exposure

Table 7.2.2/3: Male lung weights

Males

Animal

Lung weight (g)

 

number

 

 

 

 

Control

1

1.69

 

2

1.64

 

3

1.48

 

4

2.08

 

5

1.68

 

Mean

1.71

 

SD

0.22

 

 

 

Treated

11

1.94

 

12

1.92

 

13

1.84

 

14

1.98

 

15

1.67

 

Mean

1.87

 

SD

0.12

Table 7.2.2/3: Female lung weights

Females

Animal number

Lung weight (g)

 

 

 

Control

6

1.42

 

7

1.22

 

8

1.24

 

9

1.41

 

10

1.55

 

Mean

1.37

 

SD

0.14

 

 

 

Treated

16

1.62

 

17

1.65

 

18

1.48

 

19

1.51

 

20

1.53

 

Mean

1.56

 

SD

0.07

 

 

 

Interpretation of results:
GHS criteria not met
Conclusions:
The 4 hour inhalation LC0 value of the test substance was equal or higher to 5.11 mg/L.
Executive summary:

In an acute inhalation toxicity study, groups of five male and five female Sprague-Dawley rats were exposed, nose-only, to test substance concentration of 0 (control) or 5 mg/L. The mean Mass Median Aerodynamic Diameter (MMAD) value for exposure was within the guideline expected range (3.4 µm). This study was performed according to OECD Guideline 403 and with Good Laboratory Practices.

A preliminary exposure, with 1 male and 1 female rat, was conducted at the target (Limit test) concentration of 5 mg/L for a period of 4-hours in order to assess the likely response of rats to the test substance. As a concentration of 5.63 mg/L was tolerated by the preliminary investigation animals, the main study test animals were also exposed to the target concentration of 5 mg/L (concentration achieved:5.11 mg/L) for a period of 4-hours. Control animals received a single 4-hour exposure to air only.

No deaths were observed.

Immediately following exposure all test animals had reduced body temperature, whilst 7/10, 5/10 and 5/10 animals had irregular breathing, hunched posture and post dose salivation staining, respectively. In addition chin rubbing, unsteady gait and partially closed eyelids were recorded in 4/10, 3/10 and 2/10 animals, respectively. These signs generally persisted to 1 and 2-hours post-exposure, however with decreasing incidence, and were no longer present from Day 2 onwards.

Ungroomed coat and noisy breathing/râles were recorded at 2-hours post-exposure dose in 10/10 and 6/10 animals respectively, and persisted up to Days 3 and 5 respectively, however with decreasing incidence over time. In addition gasping and underactive behaviour were noted in 3/10 and 1/10 animals respectively at 2-hours post-exposure, whilst distended abdomen was recorded in one male and one female; these signs were not present at Day 2, with the exception of one male which had a distended abdomen and was gasping up to Day 2/3. Piloerection, recorded in all test animals, was generally observed from 2-hours post-exposure, and persisted up to Day 4.

Mean lung weights were slightly higher in treated animals (1.09X and 1.14X control, in male and females, respectively).

The macroscopic examination performed fourteen days after exposure did not reveal any treatment-related changes. The nature and incidence of all the findings were consistent with the commonly seen background macroscopic changes.

The 4 -hour inhalation LC50 was found to be greater than 5.11 mg/L (ie 5110 mg/m3)

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
5 110 mg/m³
Quality of whole database:
Study complete and sufficient to fulfill the endpoint requirements.

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 2010-09-28 to 2010-10-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: 8 weeks old
- Weight at study initiation: 322 ± 9g for the males and 212 ± 11g for the females
- Fasting period before study: the animals were fasted during the night before treatment but had free access to water.
- Housing: The animals were housed in polycarbonate cages with stainless steel lid. During the acclimation period, each cage (43 cm x 22 cm x 20 cm) contained one to seven animals of the same sex. During the treatment period, the animals were housed individually in smaller cages (35.5 cm x 23.5 cm x 19.3 cm).
- Diet (e.g. ad libitum): ad libitum (SSNIFF R/M-H pellet diet)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50± 20 %
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: no data
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 10% of body surface, dorsal site
- Type of wrap if used: hydrophilic gauze pad + adhesive hypoallergenic aerated semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Removal of dressing: 24h post-exposure
- Washing: at 24h post-exposure, with a moistened cotton pad

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw. The quantity of test item applied to each animal was adjusted according to the body weight recorded on the day of dose application.
- Constant volume or concentration used: no
- For solids, paste formed: The test item was placed on an area on the hydrophilic gauze pad pre-moistened with 2 mL of purified water, which was then applied to the skin.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Ten Sprague-Dawley rats (five males and five nulliparous and non pregnant females).
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment on day 1; then on days 8 and 15.
- Necropsy of survivors performed: yes
Statistics:
None
Key result
Sex:
male/female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths were observed during the study.
Clinical signs:
No clinical signs were observed during the study.
Body weight:
When compared to laboratory historical control data, a lower body weight gain was noted in 2 out of 5 males all over the observation period (34 and 26 g vs. 47 ± 7 g in control database between days 1 and 8 then 38 and 41 g vs. 51 ± 8 g in control database between days 8 and 15) and in another male between day 1 and 8 (34 g vs. 47 ± 7 g in control database). The body weight gain of the last returned to normal thereafter. The body weight of the other animals was not affected by the treatment with the test substance.
Gross pathology:
At necropsy, no apparent abnormalities were observed in any animals.
Other findings:
Erythema was noted in 1 out 5 males on day 2 only.

 

 

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD0 of the test substance was equal or higher than 2000 mg/kg bw.
Executive summary:

In a dermal acute toxicity study, the toxicity of the test substance was evaluated following a single dermal application to Sprague-Dawley rats according to OECD No. 402 guideline. The study was conducted in compliance with the principles of Good Laboratory Practice.

The test substance was applied in its original form for 24 hours to the skin of five males and five females Sprague-Dawley rats at the dose level of 2000 mg/kg bw. Then the site of application was covered by a semi-occlusive dressing.

Each animal was observed at least once a day for mortality and clinical signs for a period of up to 14 days following the single administration. From day 2, any local reaction at the treatment site was noted. The body weight of the animals was recorded on the day of treatment and then on days 8 and 15 for the surviving animals.

At the end of the observation period, all surviving animals were sacrified by carbon dioxide asphyxiation and a macroscopic post-mortem examination was performed on all animals.

No deaths and no clinical signs were observed during the study. Erythema was noted in 1 out of 5 males on day 2 only.

When compared to laboratory historical control data, a lower body weight gain was noted in 2 out of 5 males all over the observation period and in another male between days 1 and 8. The body weight gain of the last returned to normal thereafter. The body weight of the other animals was not affected by the treatment with the test substance.

At necropsy, no apparent abnormalities were observed in any animals.

The acute dermal LD50 was found greater than 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Study complete and sufficient to fulfill the endpoint requirements.

Additional information

Oral Route:

In an acute oral toxicity study, performed according to OECD 423 guideline and in compliance with Good Laboratory Practices, groups of 3 fasted Sprague-Dawley female rats were administered a single oral dose of  the test substance in 0.5 % methylcellulose by gavage. The animals were observed for mortality, clinical signs and body weight for 14 days and then necropsied for macroscopic observations.

Three animals were first given an oral dose of 300 mg/kg bw. As no mortality nor clinical signs were observed, a higher dose was tested. Therefore two independent and successive groups of 3 female rats each were given the maximum dose of 2000 mg/kg bw. Again, no mortality nor clinical signs were observed at 2000 mg/kg bw. A lower body weight gain was noted in 1/6 tested females between days 1 and 8 but the body gain returned to normal thereafter. At necropsy, one female treated with 2000 mg/kg bw presented a dilatation of both horns of uterus accompagnied of a translucent content. Since no apparent abnormalities were observed in the other animals at the both given doses, these effects were considered not significant.

The acute oral LD0 was found equal or greater than 2000 mg/kg bw.

Inhalation Route:

In an acute inhalation toxicity study, an aerosol of the test substance was tested in Sprague-Dawley rats (5/sex), by nose only way of exposure at nominal concentrations of 0 (as control) or 5 mg/mL. The mean Mass Median Aerodynamic Diameter (MMAD) value for exposure was within the guideline expected range (3.4 µm). A preliminary exposure, with 1 male and 1 female rat, was conducted at the target concentration of 5 mg/L for a period of 4-hours in order to assess the likely response of rats to the test substance.This study was performed according to OECD Guideline 403 (Limit test) and with Good Laboratory Practices.

According to results from the preliminary exposure, the main study test animals were exposed to the target concentration of 5 mg/L (concentration achieved: 5.11 mg/L) for a period of 4-hours. Control animals received a single 4-hour exposure to air only.

No deaths were observed.

Immediately following exposure all test animals had reduced body temperature, whilst 7/10, 5/10 and 5/10 animals had irregular breathing, hunched posture and post dose salivation staining, respectively. In addition chin rubbing, unsteady gait and partially closed eyelids were recorded in 4/10, 3/10 and 2/10 animals, respectively. These signs generally persisted to 1 and 2-hours post-exposure, however with decreasing incidence, and were no longer present from Day 2 onwards.

Ungroomed coat and noisy breathing were recorded at 2-hours post-exposure dose in 10/10 and 6/10 animals respectively, and persisted up to Days 3 and 5 respectively, however with decreasing incidence over time. In addition gasping and underactive behaviour were noted in 3/10 and 1/10 animals respectively at 2-hours post-exposure, whilst distended abdomen was recorded in one male and one female. These signs were not present at Day 2, with the exception of one male which had a distended abdomen and was gasping up to Day 2/3. Piloerection, recorded in all test animals, was generally observed from 2-hours post-exposure, and persisted up to Day 4.

Mean lung weights were slightly higher in treated animals. The macroscopic examination performed fourteen days after exposure did not reveal any treatment-related changes. The nature and incidence of all the findings were consistent with the commonly seen background macroscopic changes.

The 4 -hour inhalation LC50 was found to be greater than 5.11 mg/L (ie 5110 mg/m3)

Dermal route:

In a dermal acute toxicity study, the toxicity of the test substance was evaluated following a single dermal application to Sprague-Dawley rats according to OECD 402 guideline. The study was conducted in compliance with the principles of Good Laboratory Practices.

The test substance was applied in its original form for 24 hours to the skin of five males and five females Sprague-Dawley rats at the dose level of 2000 mg/kg bw. Then the site of application was covered by a semi-occlusive dressing.

Each animal was observed at least once a day for mortality and clinical signs for a period of up to 14 days following the single administration. From day 2, any local reaction at the treatment site was noted. The body weight of the animals was recorded on the day of treatment and then on days 8 and 15 for the surviving animals. At the end of the observation period, all surviving animals were sacrificed by carbon dioxide asphyxiation and a macroscopic post-mortem examination was performed on all animals.

No deaths and no clinical signs were observed during the study. Erythema was noted in 1/5 males on day 2 only. When compared to historical control data, a lower body weight gain was noted in 2/5 males all over the observation period and in another male between day 1 and 8. The body weight gain of the last returned to normal thereafter. The body weight of the other animals was not affected by the treatment . At necropsy, no apparent abnormalities were observed in any animals.

The acute dermal LD50 was found greater than 2000 mg/kg bw.


Justification for classification or non-classification

No mortalities were noted in rats either after treatment by oral or dermal routes with a single dose of 2000 mg/kg body weight or exposure by inhalation for 4 hours to 5.11 mg/L.

On the basis of these results and according to regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures, no classification is warranted with respect to acute oral, dermal and inhalation toxicities.