4-(5-(3,5-dichloro-4-fluorophenyl)-5-(trifluoromethyl)-4,5-dihydro-1,2-oxazol-3-yl)-N-(2-ethyl-3-oxo-1,2-oxazolidin-4-yl)-2-methylbenzamide

Endpoint summary

Description of key information

- Oral: LD50 > 5000 mg/kg bw, female, rat, according to OECD 425, Tarcai 2016

- Inhalation: LC50 > 4.62 mg/L, males/female, rats, according to OECD 403, Rosos-Mating 2017

- Dermal: LD50 > 5000 mg/kg bw, male/female, rat, according to OECD 402, Tarcai, 2016

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:

8 Mar 2016 to 5 Apr 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

Version / remarks:

2018

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

2008

GLP compliance:

yes (incl. QA statement)

Test type:

up-and-down procedure

Limit test:

no

Species:

rat

Strain:

Wistar

Remarks:

Crl:(WI)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Sex: Female rat, nulliparous and non-pregnant
- Age: Young adult rats, 8 weeks old
- Body weight when treated: 199 – 216 g
- Fasting period: animals were fasted overnight prior to treatment and food was returned approximately 3 hours after dosing
- Diet: Animals received "Autoclavable complete diet for rats and mice – breeding and maintenance" ad libitum
- Water: Tap water from municipal supply, provided in 500 mL bottles ad libitum
- Housing: Individual caging Type II. polypropylene/polycarbonate cages. Laboratory bedding were available to animals during the study
- Acclimatisation period: At least 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.0 – 21.9
- Humidity (%): 26 – 70
- Air changes (exchanges/hour): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: from 08 Mar 2016 to 05 Apr 2016

Route of administration:

oral: gavage

Vehicle:

other: 0.5 % (w/v) carboxymethyl cellulose and 0.1 % (v/v) Tween 80

Details on oral exposure:

The applied dose volume was 10 mL/kg bw

Doses:

Starting dose of 1750 mg/kg bw, then 5000 mg/kg bw as no clinical signs or lethality were observed, followed by a 14 day observation period. Animals were treated with a single oral (gavage) dose.

No. of animals per sex per dose:

4

Control animals:

no

Details on study design:

All animals were observed individually after dosing at 30 minutes, then 1, 2, 3, 4, and 6 hours after dosing and once each day for 14 days thereafter. Individual observations were performed on the skin and fur, eyes and mucous membranes and also 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 weights were recorded on Days -1, 0 (before treatment), 7 and 14. All animals were euthanised at the end of the observation period by exsanguination under pentobarbital anaesthesia and all animals were subjected to macroscopic examination. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross pathological changes were recorded for each animal on the post mortem record sheets and the animals were discarded.

Statistics:

The oral LD50 was calculated using the statistical programme (AOT 425 Stat Pgm) version: 1.0, 2001, used for the selection of dose levels and calculation of the LD50 values.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs were observed during the observation period.

Gross pathology:

At necropsy, there were no observations to be reported at a dose level of 1750 and 5000 mg/kg bw.

Table 1: Acute oral toxicity of the test substance in rats, application scheme and mortality data

Animal Number	Dose [mg/kg body weight]	Volume Dosed [mL]	Bodyweight [g]	Mortality
1	1750	2.1	207	Survived
2	5000	2.0	199	Survived
3	5000	2.0	199	Survived
4	5000	2.2	216	Survived

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the acute oral median lethal dose (LD50) of the test substance was greater than 5000 mg/kg bw in female Crl:WI rats.

Executive summary:

An acute oral toxicity study (OECD 425, GLP compliant) (up and down procedure) was conducted with 4 female Crl:WI rats. Animals were treated with a single oral (gavage) dose of the test substance, at a dose level of 1750 or 5000 mg/kg body weight (bw) followed by a 14 day observation period. The animals were fasted overnight prior to treatment and food was returned 3 hours after dosing. Animals were observed individually after dosing at 30 minutes, then at 1, 2, 3, 4 and 6 hours post treatment and once each day for 14 days, thereafter. Body weight was measured on Day -1, just before dosing and weekly thereafter. All animals were examined macroscopically at the end of the observation period.  There was no mortality during the study. No clinical signs were observed during the observation period. There were no treatment related effects on body weight or body weight gain. Body weights were within the range commonly recorded for this strain and age. At necropsy, there were no observations to be reported at a dose level of 1750 and 5000 mg/kg bw.

Under the conditions of this study, the acute oral median lethal dose (LD50) of the test substance, the test substance, was greater than 5000 mg/kg bw in female Crl:WI rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Quality of whole database:

GLP compliant OECD 425 study.

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:

20 Jul 2016 to 17 Aug 2016

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

1998

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Version / remarks:

2008

GLP compliance:

yes

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:(WI)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 12 weeks old
- Body weight at study initiation: males: 396 - 420 g; females: 244 - 269 g
- Diet: Autoclavable Complete Feed for Rats and Mice – Breeding and Maintenance, ad libitum
- Water: Tap water, ad libitum
- Housing: Group caging in Type III solid floor cages with stainless steel mesh lids. Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities
- Acclimatisation period: At least 5 days

ENVIRONMENTAL CONDITION
- Temperature (°C): 19.0 – 24.3
- Humidity (%): 34 – 70
- Air changes (per hour): 15 - 20
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 20 Jul 2016 To:17 Aug 2016

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

>= 3.11 - <= 3.38 µm

Geometric standard deviation (GSD):

>= 2.34 - <= 2.37

Details on inhalation exposure:

EXPOSURE CONDITIONS
Prior to animal exposures, test material atmospheres were generated within the exposure chamber. During these technical trials, air-flow settings, test substance input rates were varied to achieve the required aerosol concentration of particles Measurements of aerodynamic particle size were performed from the animal’s breathing zone using a cascade impactor. Due to the physicochemical properties of the test substance, it was not possible to achieve the maximum (limit) concentration of 5 mg/L with an MMAD of 1 - 4 µm. More information is provided in a table in the section "Any other information on materials and methods incl tables"

GENERATION OF THE TEST ATMOSPHERE / CHAMBER DESCRIPTION
The test substance was aerosolised using a Wright’s Dust Feed System located at the top of the exposure chamber. Compressed air was supplied by means of an oil-free compressor and passed through a suitable filter system prior to introduction to the dust generator. The animals were exposed, nose-only, to an atmosphere of the test substance using a TSE Rodent Exposure System. This system comprised of 2 concentric anodised aluminium chambers and a computer control system incorporating pressure detectors and mass flow controllers. Fresh aerosol from the generation system was constantly supplied to the inner plenum (distribution chamber) of the exposure system from where, under positive pressure, it was distributed to the individual exposure ports. The animals were held in polycarbonate restraint tubes located around the chamber which allowed only the animal’s nares to enter the exposure port. After passing through the animal’s breathing zone, used aerosol entered the outer cylinder from where it was exhausted through a suitable filter system. Atmosphere generation was therefore dynamic. Airflows and relative pressures within the system were constantly monitored and controlled by the computer system thus ensuring a uniform distribution and constant flow of fresh aerosol to each exposure port (breathing zone). The flow of air through each port was at least 0.7 L/min. This flow rate was considered adequate to minimise re-breathing of the test atmosphere as it is about twice the respiratory minute volume of a rat. Homogeneity of the test atmosphere within the test chamber and amongst the exposure ports was not specifically determined during this study. However, chambers of this design have been fully validated and have shown to produce evenly distributed atmospheres in the animals’ breathing zones (Pauluhn, 1994).

TEST ATMOSPHERE CONCENTRATION
The test atmosphere was sampled at regular intervals during each exposure period. Samples were taken from an unoccupied exposure port (representing the animal’s breathing zone) by pulling a suitable, known volume of test atmosphere through weighed GF10 glass fibre filters. 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. The nominal concentration was calculated by dividing the mass of test material disseminated into the chamber by the total volume of air that through the chamber during the same period. Particle size determination: The particle size of the test atmosphere was determined three times during the exposure period using a 7-stage impactor of Mercer style. Such devices employ an inertial separation technique to isolate particles in the discrete aerodynamic size ranges. Samples were taken from an unoccupied exposure port (representing the animal’s breathing zone). The collection substrates and the backup filter were weighed before and after sampling and the weight of test item, collected at each stage, 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 0.55, 0.96, 1.55, 2.11, 3.56, 6.66 and 10.55 μm was calculated. From these data, using software supplied with the impactor , the Mass Median Aerodynamic Diameter (MMAD), and Geometric Standard Deviation (GSD) were calculated. In addition, the proportion (%) of aerosol less than 4 μm (considered to be the inhalable portion) was determined.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

4.76 mg/L

No. of animals per sex per dose:

Sighting exposure: 2
Main group: 5

Control animals:

no

Details on study design:

ANIMAL ASSIGNMENT AND TREATMENT
Prior to the start of the study the animals were examined to ensure that they were physically normal and exhibited normal activity. All animals were observed for mortality during the exposure period. The animals were examined for signs of gross toxicity, and behavioural changes upon removal from the exposure tube and at least once daily thereafter for 14 days. Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea and coma. In group 1, one female rat was found dead on Day 7 and was cannibalized. A specific cause of death was not determined for this animal. At necropsy collapsed, dark/red discoloured lungs were observed. No clinical observations were observed in this animal for several days prior to death and whilst the animal lost body weight following treatment, these losses were no greater than those observed in other animals in this treatment group. It is therefore uncertain that the death of this animal was a result of test item administration. Individual body weights were recorded prior to treatment on the day of exposure (Day 0) and on Days 1, 3, 7 and 14. At the end of the 14 day observation period, the surviving animals were sacrificed by exsanguination under anaesthesia (Euthanimal 40 % (pentobarbital sodium) injection and gross macroscopic examination was performed. All animals were subject to a gross necropsy which included a detailed examination of the abdominal and thoracic cavities. Special attention was given to the respiratory tract for macroscopic signs of irritancy or local toxicity.

Statistics:

The acute inhalation LC50 was not estimated.

Preliminary study:

During the sighting study, laboured respiration (slight) was observed. The animals were symptom free from Day 1

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

4.62 mg/L air (analytical)

Exp. duration:

4 h

Remarks on result:

other: Highest attainable concentration

Mortality:

In group 1, one female rat was found dead on Day 7 and was cannibalized. A specific cause of death was not determined for this animal

Clinical signs:

other: Ruffled fur or red-brown staining (as chromodacryorrhea) and fur staining by the test substance occurring in study animals from Day 0 - 2 were considered to be related to the restraint and exposure procedures not to be toxicologically significant.

Remarks:

During the sighting and the main study, laboured respiration (slight) was observed. The animals were symptom free from Day 1

Body weight:

In Group 0.1, slight and moderate body weight loss were observed in both sexes between Day 0 and 3 (males: 5.9 - 7.3 %, females: 13.7 - 17 % respectively) of the observation period. The body weight gain of male and female rats normalised by the end of the observation period. In Group 1, all male and all female animals showed slight or moderate body weight loss (males: 4.8 - 10 %, females: 11.7 - 13.9 % respectively) in the first 3 days of the observation period. In case of surviving animals, the body weight gain returned to the normal range, thereafter (Day 14).

Gross pathology:

A single four hour nose-only exposure of the test substance to Crl: WI Wistar strain rats exposed to the concentration of 4.38 and 4.62 mg/L during Sighting exposure and Main study, respectively, did not produce any test substance-related gross findings on Day 14.

Table 3: Mortality overview

Group Number	Mean Achieved (mg/L)	Male Deaths	Female Deaths	Total Deaths
1	(Maximum attainable)	0/5	1/5	1/10

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, one female was found dead in a group of 10 rats exposed to the mean achieved concentration of 4.62 mg/L (maximum attainable concentration) for 4 hours. The acute inhalation median lethal concentration of the test substance, in CRL:WI Wistar strain rats is therefore considered to be greater than 4.62 mg/L

Executive summary:

This OECD 403 study performed in compliance with GLP to assess the acute inhalation toxicity of the test substance following a 4 hour exposure at the maximum achievable concentration for the test substance was approximately 4.76.mg/L to 5 male and 5 female rats. A sighting exposure was performed prior to the main study with 2 males and 2 females at a mean achieved concentration of 4.38 mg/L. The main study group, 10 (5 male and 5 female) CRL:WI Wistar strain rats, was exposed to the mean achieved concentration of 4.62 mg/L the test substance The animals were exposed for 4 hours using a nose-only exposure system, followed by a 14 day observation period. The day of exposure was designated Day 0. Aerosol concentrations were measured gravimetrically. The particle size distribution of the test aerosol was determined regularly during the exposure period. Clinical observations and body weights were recorded throughout the study and at the end of the scheduled period the animals were euthanised and subjected to a gross examination post mortem.

In group 1, one female rat was found dead on Day 7 and was cannibalized.  A specific cause of death was not determined for this animal.  At necropsy collapsed, dark/red discoloured lungs were observed. Ruffled fur or red-brown staining (as chromodacryorrhea) and fur staining by the test substance occurring in study animals from Day 0 up to Day 2 were considered to be related to the restraint and exposure procedures not to be toxicologically significant. During the sighting and the main study, laboured respiration (slight) was observed. The animals were symptom free from Day 1. In Group 0.1, slight and moderate body weight loss were observed in both sexes between Day 0 and 3 (males: 5.9 - 7.3 %, females: 13.7 - 17 % respectively) of the observation period. The body weight gain of male and female rats normalised by the end of the observation period.

In Group 1, all male and all female animals showed slight or moderate body weight loss (males: 4.8 - 10 %, females: 11.7 - 13.9 % respectively) in the first 3 days of the observation period. In case of surviving animals, the bodyweight gain returned to the normal range, thereafter (Day 14). A single four hour nose-only exposure of the test substance to Crl: WI Wistar strain rats exposed to the concentration of 4.38 and 4.62 mg/L during Sighting exposure and Main study, respectively, did not produce any test substance-related gross findings on Day 14. Under the experimental conditions of this study, one female was found dead in a group of 10 rats exposed to the mean achieved concentration of 4.62 mg/L (maximum attainable concentration) for 4 hours. The acute inhalation median lethal concentration of the test substance, in CRL:WI Wistar strain rats is therefore considered to be greater than 4.62 mg/L.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

4 620 mg/m³ air

Quality of whole database:

GLP compliant OECD 403 study.

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:

8 Mar 2016 to 24 Mar 2016

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:

February 1987

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

August 1998

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No 440/2008, B.3 (L 142)

Version / remarks:

May 2008

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:(WI) Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Female rats were nulliparous and nonpregnant
- Age at study initiation: Young adult rats
- Weight at study initiation: 212 - 251 g
- Identification: Animals were identified by numbers written on the tail with an indelible marker. The cages were marked with individual identity cards with information about study number, sex, cage number, dose group and individual animal number.
- Housing: Animals were housed individually in Type II polypropylene/polycarbonate cages. Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
- Randomization: Selected by hand at time of delivery
- Acclimatization time: 5 or 7 days
- Diet: Autoclavable complete diet for rats and mice – breeding and maintenance, ad libitum
- Water: tap water, from the municipal supply, provided in 500 mL bottles ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 – 24.4
- Humidity (%): 31 - 61
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 08 Mar 2017 To: 24 Mar 2017

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

The back of the animal was shaven (approximately 10 % area of the total body surface) approximately 24 hours prior to the treatment. Only those animals without injury or irritation on the skin were used in the test.
On test Day 0, the test substance was applied as a single dose of 5000 mg/kg body weight, moistened with water, and distributed as uniformly as possible over the skin and remained on the skin throughout a 24-hour exposure period. Sterile gauze pads were placed on the skin of rats at the site of application. These gauze pads were kept in contact with the skin by 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, residual testsubstance was removed, using body temperature water

Duration of exposure:

24 hours

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

EXPERIMENTAL DESIGN
A single administration was performed by the dermal route and was followed by a 14-day observation period.
One male and one female sentinel rats were dosed initially and the remaining four male and four female rats were dosed 48 hours later when it was clear there were no adverse effects.

CLINICAL OBSERVATIONS
A clinical examination was performed on the day of treatment, at 1 and 5 hours after the application of the test item, and once each day for 14 days thereafter.
Observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behaviour pattern. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

SKIN IRRITATION
Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

MEASUREMENT OF BODY WEIGHT
The body weight of all animals was recorded on Day 0 (day of treatment), and on Days 7 and 14.

POST MORTEM INVESTIGATIONS
All animals were subjected to gross macroscopic examination. All animals were anaesthetised with Euthanimal 40% (Pentobarbital sodium 400 mg/mL) and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. Any gross macroscopic findings were recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: There were no adverse clinical signs noted in any animals throughout the study.

Gross pathology:

There was no evidence of the test item-related observations at a dose level of 5000 mg/kg bw at necropsy.

Other findings:

No treatment related skin irritation was observed in any animal throughout the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose of the test substance after a single dermal administration was found to be greater than 5000 mg/kg bw in male and female CRL:(WI) Wistar rats.

Executive summary:

In a OECD 402 study, in compliance with GLP, five male and five female Crl:WI rats were treated with a single semi-occlusive dermal application of test substance at the limit dose of 5000 mg/kg bw. The application period was 24 hours, followed by a 14-day observation period. Clinical observations along with a check of viability and mortality were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. All animals were euthanised and subjected to a gross macroscopic examination at the end of the 2 week observation period (Day 14).

No mortality occurred during the study. No adverse clinical signs were observed after treatment with the test substance or during the 14 day observation period and no effects were observed at the site of application. There were no treatment related body weight changes. Body weights for the animals were within the range commonly recorded for this strain and age. No macroscopic changes were observed at necropsy. The median lethal dose (LD50) of test substance after a single dermal administration was greater than 5000 mg/kg bw in male and female Crl:WI rats

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Quality of whole database:

GLP compliant OECD 402 study.

Additional information

Acute toxicity: oral

An acute oral toxicity study (OECD 425, GLP compliant) (up and down procedure) was conducted with 4 female Crl:WI rats. Animals were treated with a single oral (gavage) dose of the test substance, at a dose level of 1750 or 5000 mg/kg body weight (bw) followed by a 14 day observation period. The animals were fasted overnight prior to treatment and food was returned 3 hours after dosing. There was no mortality during the study. No clinical signs were observed during the observation period. There were no treatment related effects on body weight or body weight gain. At necropsy, there were no observations to be reported at a dose level of 1750 and 5000 mg/kg bw. Under the conditions of this study, the acute oral LD50 value was greater than 5000 mg/kg bw in female Crl:WI rats.

Acute toxicity: inhalation

An acute inhalation toxicity study (OECD 403, GLP compliant) was performed with 5 male and 5 female rats exposed (nose only) for a period of 4 hours to the maximum achievable aerosol concentration of approximately 4.62 mg/L. The exposure was followed by a 14 day observation period. Aerosol concentrations were measured gravimetrically. The particle size distribution of the test aerosol was determined regularly during the exposure period. One female rat was found dead on Day 7.  A specific cause of death was not determined for this animal, but collapsed, dark/red discoloured lungs were observed at necropsy. Ruffled fur or red-brown staining (as chromodacryorrhea) and fur staining by the test substance occurring in study animals from Day 0 up to Day 2 were considered to be related to the restraint and exposure procedures and not to be toxicologically significant. Slightly laboured respiration was observed. The animals were symptom free from Day 1. All male and all female animals showed slight or moderate body weight loss (males: 4.8 - 10%, females: 11.7 - 13.9% respectively) in the first 3 days of the observation period. The body weight gain returned to the normal range, thereafter (Day 14). No test substance-related gross were found at the end of the observation period. Under the experimental conditions the acute inhalation median lethal concentration of the test substance, in CRL:WI Wistar strain rats was considered to be greater than 4.62 mg/L.

Acute toxicity: dermal

An acute dermal toxicity study (OECD 402, GLP compliant) was performed with five male and five female Crl:WI rats receiving a single semi-occlusive dermal application of test substance at the limit dose of 5000 mg/kg bw. The application period was 24 hours, followed by a 14-day observation period. No mortality occurred during the study. No adverse clinical signs were observed after treatment with the test substance or during the 14 day observation period and no effects were observed at the site of application. There were no treatment related body weight changes. No macroscopic changes were observed at necropsy. The median lethal dose (LD50) of test substance after a single dermal administration was greater than 5000 mg/kg bw in male and female Crl:WI rats

Justification for classification or non-classification

Based on the results of the acute toxicity studies (oral, dermal, inhalation), classification for acute toxicity is not warranted in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.