(2E)-2-methyl-3-(4-methylphenyl)prop-2-en-1-ol

Endpoint summary

Description of key information

Oral: measured LD50 > 300 and < 2000 mg/kg bw and the estimated LD50 cut-off was considered to be 2000 mg/kg bw, female rat, OECD TG 423, 2012

Inhalation: LC50 (male/female): > 5.1 mg/L, mean maximum achievable atmosphere concentration, male/female rat, OECD TG 403, 2012

Dermal: measured LD50 > 2000 mg/kg bw and the estimated LD50 cut-off value was considered to be > 5000 mg/kg bw, male/female rat, OECD TG 402, 2012

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:

08-09-2011 to 07-10-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Justification for type of information:

Information as to the availability of the in vivo study is provided in 'attached justification'.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: 8 - 9 weeks (nulliparous and non-pregnant)
- Weight at study initiation: 142 - 195 g; Body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: Overnight and until 3-4 hours after administration of the test substance.
- Housing: Group housing of 3 animals per cage containing sterilized sawdust as bedding material and paper as cage-enrichment.
- Water: ad libitum
- Acclimation period: At least five (5) days under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual: 19.0 to 22.6°C)
- Humidity (%): 40 to 70 (actual: 44 – 82%)
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 hours light/dark

IN-LIFE DATES: 08-09-2011 to 07-10-2011

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Identity: Polyethylene glycol 400
- Concentration in vehicle: The concentration of the test item in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION (if unusual): The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Since the test substance was in a fluid state at the time of weighing, it was not heated.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: In the absence of data suggesting the test material was toxic, 2000 mg/kg was chosen as 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. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

Doses:

300 mg/lg and 2000 mg/kg

No. of animals per sex per dose:

6

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Mortality/Viability: Twice daily; Bodyweights: 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. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded.
- Necropsy of survivors performed: yes

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

2000 mg/kg bw: there was 2/6 mortalities (2/3 in the second group.
300 mg/kg bw: there was no mortalities.

Clinical signs:

other: 2000 mg/kg bw: Lethargy, flat and/or hunched posture, piloerection, slow breathing, uncoordinated movements, dark eye and/or watery discharge from the eyes were noted in all animals between Days 1 and 6. One animal showed scales and scabs on the snout dur

Gross pathology:

No abnormalities were found at macroscopic post mortem examination.

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the oral LD50 established to be > 300 and < 2000 mg/kg bw in female Wistar rats. According to OECD TG 423 the LD50 cut-off value was considered to be 2000 mg/kg bw.

Executive summary:

The study was performed according to OECD TG 423 and EU Method B1 tris Acute Toxicity, US EPA OPPTS 870.1100 and Japanese JMAFF guidelines and in accordance with GLP to assess the acute oral toxicity of the test item following a single oral administration in the female Wistar strain rat by the acute class method. The test item was administered by oral gavage in a PEG 400 vehicle to two sequential groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). There was two mortalities in the second group. Lethargy, flat and/or hunched posture, piloerection, slow breathing, uncoordinated movements, dark eye and/or watery discharge from the eyes were noted in all animals between Days 1 and 6. In addition one animal showed scales and scabs on the snout during the observation period. A second dose level was then employed at 300 mg/kg bodyweight. All animals showed hunched posture and one animal showed piloerection on Day 1. No further effects were subsequently seen. The body weight gain shown by survivors over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals at any dose level. The oral LD50 value of the test substance in Wistar rats was established to be > 300 and < 2000 mg/kg body weight. According to the OECD TG 423 test guideline, the LD50 cut-off value was considered to be 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

300 mg/kg bw

Quality of whole database:

The available information as a whole meets the tonnage driven information requirements of REACH.

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:

21-05-2013 to 04-06-2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Justification for type of information:

Information as to the availability of the in vivo study is provided in 'attached justification'.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

inspected: March 2013 ; signature: May 2013

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Recognised supplier
- Age at study initiation: 10 - 11 weeks
- Weight at study initiation: Approximately the same age and body weight variation did not exceed +/- 20% of the sex mean. Females: 194-224 g and Males: 310 – 325 g
- Fasting period before study: None.
- Housing: Group housing of five animals per sex per cage in labelled Makrolon cages (type IV), containing sterilised sawdust bedding and paper cage enrichment.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for exposure period period)
- Water (e.g. ad libitum): mains drinking water, ad libitum (except for exposure period period)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

IN-LIFE DATES: From: 21-05-2013 To: 04-06-2013

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

>= 3.3 - <= 3.4 µm

Geometric standard deviation (GSD):

>= 1.9 - <= 2

Remark on MMAD/GSD:

MMAD/GSD relates to: 6.9 mg/L (nominal), 5.1 ± 0.1 mg/L (mean achieved concentration) dose - determined twice during the study

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: An aerosol was generated by nebulization of the test substance by means of a nebulizer (type 950) and pressurized air. The nebulizer was placed in a water bath of approximately 40 ºC which was placed on a magnetic stirrer. The primary aerosol was diluted with humidified pressurized air before it entered the exposure chamber
- Exposure chamber volume: Not reported. Was consistent with Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983.
- Method of holding animals in test chamber: Restraining tubes.
- Source and rate of air: filtered air
- Method of conditioning air: Compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters before it was introduced to the nebulizer.
- System of generating particulates/aerosols: nebulizer; the chamber mean total flow rate was maintained at 18 L/min.
- Method of particle size determination: Particle size was determined using a cascade impactor. The device consisted of eight impactors stages containing fiber glass filters.
- Treatment of exhaust air: From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.
- Temperature, humidity, in air chamber: The temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter: 27.6-29.1°C, 87-91% humidity.

TEST ATMOSPHERE
- Brief description of analytical method used: Actual concentration was determined sixteen 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. 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. The time-weighted mean concentration with the standard deviation was calculated. Full details of the analytical method are provided in the full study report.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: The particle size of the generated atmosphere inside the exposure chamber was determined two times during each exposure period using a cascade impactor. The particle size distribution for each group is reported in table 1.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The Mass Median Aerodynamic Diameter (MMAD) was determined and is reported for each group in table 1.
stribution for each group is reported in table 1.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

6.9 mg/L (nominal), 5.1 mg/L (mean achieved concentration) with a generation efficiency of 74%.

No. of animals per sex per dose:

5 per sex per dose. Full details are provided in table 2.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Mortality, twice daily. Clinical signs three times during exposure and on day one at one and three hours and then once daily. Any evidence of mortality or overt toxicity was recorded at each observation. Individual body weights were recorded on arrival, prior to treatment on the day of exposure and on Days 1, 4, 8 and 15 or after mortality.
- Necropsy of survivors performed: yes (and in the event of any mortalities)
- Other examinations performed: clinical signs, body weight, organ weights, and any other relevant toxicological effects were reported.

Statistics:

No statistical analysis was performed.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.1 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

1 female mortality occurred on day 1 ; mortalities are reported in table 3.

Clinical signs:

other: No clinical signs were noted during exposure. After exposure, the clinical signs observed among the animals were lethargy, tremors, flat posture, hunched posture, uncoordinated movements, slow breathing, laboured respiration, shallow respiration, rales, g

Body weight:

All animals exhibited body weight losses on the first week post-exposure. Body weight gains were noted in all surviving animals during the remainder of the recovery period.

Gross pathology:

In the female mortality: macroscopic post mortem examination of the female found dead revealed light red discoloration of the lungs.
In the survivors: cloudy eyes were observed. No other abnormalities were found at macroscopic examination.

Other findings:

- Other observations: The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity during necropsy.

Additional comments regarding test atmosphere generation:

It is noted within the study that the time-weighted mean actual concentration was 5.1 ± 0.1 mg/L and that the generation efficiency was 74%. The concentration measurements equally distributed over time demonstrated that the atmosphere was sufficiently stable at the mean total airflow of 18 L/min. Full details are available within the full study report.

 

Table 1. Characteristics of the achieved atmosphere:

Group Number	Mean Achieved Atmosphere Concentration (mg/L)	Mean Mass Median Aerodynamic Diameter (µm)	Geometric Standard Deviation	Comments
1	5.1	> 3.3 - < 3.4	> 1.9 - < 2.0	MMAD of 10% of atmosphere was < 0.5 µm

 

Table 2. Mean achieved atmosphere concentrations:

Group Number	Mean Achieved Atmosphere Concentration (mg/L)	Standard Deviation	Nominal (mg/L)
1	5.1	0.115	6.9

 

Table 3. Mortality data summary:

Group Number	Mean Achieved Atmosphere Concentration (mg/L)	Mortalities
		Female	Male	Total
1	5.1	1/5	0/5	1/10

Interpretation of results:

not classified

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the study, the inhalation 4h-LC50 (male/female) was considered to be > 5.1 mg/L within the Crl:WI(Han) rat.

Executive summary:

The study was performed according to OECD TG 403, EU Method B.2, US EPA OPPTS 870.1300 and Japanese JMAFF guidelines in accordance with GLP to assess the acute inhalation toxicity of the test item. A single group of ten Wistar: Crl:WI(Han) strain rats (five males and five females) were exposed to an aerosol atmosphere of the test item. The groups were exposed for four hours using a nose only exposure system, followed by a fifteen day observation period. The mean achieved atmosphere concentrations were as follows: 5.1 ± 0.1 mg/L based on a nominal concentration of 6.9 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size): > 3.3 μm and < 3.4 μm with geometric Standard Deviation > 1.9 and < 2.0. There was no male and one female mortalities in the 5.1 mg/L mean achieved atmosphere concentration. The single female mortality occurred within 4 hours post-exposure.No clinical signs were noted during exposure. After exposure, the clinical signs observed among the animals were lethargy, tremors, flat posture, hunched posture, uncoordinated movements, slow breathing, laboured respiration, shallow respiration, rales, gasping, piloerection, chromodacryorrhoea and/or ptosis. The surviving animals had recovered from the signs between Days 5 and 6, except for the rales in one male that were also present on Day 8. Body weight loss was noted in all animals during the first week post-exposure. All animals regained weight during the second week. Macroscopic post mortem examination of the female found dead revealed light red discoloration of the lungs. Cloudy eyes were seen in two surviving males. No other abnormalities were found at macroscopic examination. Under the conditions of this study, the inhalation 4h-LC50 (male/female) was considered to be > 5.1 mg/L within the Wistar: Crl:WI(Han) rat.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

5 100 mg/m³ air

Quality of whole database:

The available information as a whole meets the tonnage driven information requirements of REACH.

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:

06-10-2011 to 20-10-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Justification for type of information:

Information as to the availability of the in vivo study is provided in 'attached justification'.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Recognised supplier
- Age at study initiation: approximately 10 weeks.
- Weight at study initiation: Female: 194 - 210 g ; Male: 292 – 312 g ; The weight variation did not exceed +/- 20% of the mean weight per sex at the start of treatment.
- Fasting period before study: Not applicable.
- Housing: Individually housed in Makrolon cages furnished with sterilized sawdust and paper cage enrichment.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual: 19.4 to 21.2°C)
- Humidity (%): 40 to 70 (actual: 44 – 73%) - deviation had no impact on the study
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 hours light/dark

IN-LIFE DATES: 06-10-2011 to 20-10-2011

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Remarks:

The test item was a liquid at the time of dosing and could be applied as such

Details on dermal exposure:

TEST SITE
- Area of exposure: The day before treatment the back and flanks were clipped free of hair. Dorsal area application.
- % coverage: Approximately 10% of total body surface
- Type of wrap if used: The area of application was covered piece of surgical gauze was placed over the treatment area and occluded with a piece of aluminium foil and elastic bandage affixed with micropore tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair wiped with cotton wool moistened with water to remove any residual test item
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): dose volume: 1.98 mL/kg bodweight.
- Concentration (if solution): 2000 mg/kg
- Constant volume or concentration used: Not applicable.

VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 per sex per dose (5 male/5 female)

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Clinical observations and mortality checks were conducted at approximately 0.5, 1, 2, and 4 hours and subsequently once daily for 15 days. Local effects were examined once daily for 15 days after the completion of the 24-hour exposure period. Full details on the scoring and criteria are given in the full study report. Individual bodyweights were recorded prior to application of the test item on Day 0 and on Days 8 and 15.
- Necropsy of survivors performed: yes

Statistics:

No statistical analyses were performed.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

other: - Clinical observations: No signs of systemic toxicity were noted during the observation period. Chromodacryorrhoea (Snout) (score = 1) was noted in two females on day 1. - Dermal reactions: Focal erythema (score = 1) (Days 2 and 3) and scales (score = 1)

Gross pathology:

No abnormalities were noted at necropsy.

Interpretation of results:

not classified

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Executive summary:

The study was performed according to OECD TG 402, EU Method B.3 Acute Toxicity (Dermal), US EPA OPPTS 870.1100 and Japanese JMAFF guidelines in accordance with GLP to assess the acute dermal toxicity of the test item in the Wistar strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, occluded dermal application of the undiluted test item to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. Clinical signs and/or local effects consisted of chromodacryorrhoea in two females on Day 1. Whereas, focal erythema (Days 2 and 3) and scales (between Days 4-7) were observed on the treated skin-area of two males and one female. The body weight gain during the observation period was within the expected range. Under the conditions of this study, the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

The available information as a whole meets the tonnage driven information requirements of REACH.

Additional information

ORAL: Key data: OECD TG 420, 2012 - The study was performed according to OECD TG 423 and EU Method B1 tris Acute Toxicity, US EPA OPPTS 870.1100 and Japanese JMAFF guidelines and in accordance with GLP to assess the acute oral toxicity of the test item following a single oral administration in the female Wistar strain rat by the acute class method. The test item was administered by oral gavage in a PEG 400 vehicle to two sequential groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). There was two mortalities in the second group. Lethargy, flat and/or hunched posture, piloerection, slow breathing, uncoordinated movements, dark eye and/or watery discharge from the eyes were noted in all animals between Days 1 and 6. In addition one animal showed scales and scabs on the snout during the observation period. A second dose level was then employed at 300 mg/kg bodyweight. All animals showed hunched posture and one animal showed piloerection on Day 1. No further effects were subsequently seen. The body weight gain shown by survivors over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals at any dose level. The oral LD50 value of the test substance in Wistar rats was established to be > 300 and < 2000 mg/kg body weight. According to the OECD TG 423 test guideline, the LD50 cut-off value was considered to be 2000 mg/kg body weight.

 

INHALATION: Key data: OECD TG 403, 2013 - The study was performed according to OECD TG 403, EU Method B.2, US EPA OPPTS 870.1300 and Japanese JMAFF guidelines in accordance with GLP to assess the acute inhalation toxicity of the test item. A single group of ten Wistar: Crl:WI(Han) strain rats (five males and five females) were exposed to an aerosol atmosphere of the test item. The groups were exposed for four hours using a nose only exposure system, followed by a fifteen day observation period. The mean achieved atmosphere concentrations were as follows: 5.1 ± 0.1 mg/L based on a nominal concentration of 6.9 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size): > 3.3 μm and < 3.4 μm with geometric Standard Deviation > 1.9 and < 2.0. There was no male and one female mortalities in the 5.1 mg/L mean achieved atmosphere concentration. The single female mortality occurred within 4 hours post-exposure.No clinical signs were noted during exposure. After exposure, the clinical signs observed among the animals were lethargy, tremors, flat posture, hunched posture, uncoordinated movements, slow breathing, laboured respiration, shallow respiration, rales, gasping, piloerection, chromodacryorrhoea and/or ptosis. The surviving animals had recovered from the signs between Days 5 and 6, except for the rales in one male that were also present on Day 8. Body weight loss was noted in all animals during the first week post-exposure. All animals regained weight during the second week. Macroscopic post mortem examination of the female found dead revealed light red discoloration of the lungs. Cloudy eyes were seen in two surviving males. No other abnormalities were found at macroscopic examination. Under the conditions of this study, the inhalation 4h-LC50 (male/female) was considered to be > 5.1 mg/L within the Wistar: Crl:WI(Han) rat.

DERMAL: Key data: OECD TG 402, 2012 - The study was performed according to OECD TG 402, EU Method B.3 Acute Toxicity (Dermal), US EPA OPPTS 870.1100 and Japanese JMAFF guidelines in accordance with GLP to assess the acute dermal toxicity of the test item in the Wistar strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, occluded dermal application of the undiluted test item to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. Clinical signs and/or local effects consisted of chromodacryorrhoea in two females on Day 1. Whereas, focal erythema (Days 2 and 3) and scales (between Days 4-7) were observed on the treated skin-area of two males and one female. The body weight gain during the observation period was within the expected range. Under the conditions of this study, the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Applicant assessment indicates, under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Justification for classification or non-classification

The substance meets classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: oral: category 4: H302

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: inhalation

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: dermal