N-[[3,5-dichloro-2-fluoro-4-(1,1,2,3,3,3-hexafluoropropoxy)phenyl]carbamoyl]-2,6-difluorobenzamide

Endpoint summary

Description of key information

Oral toxicity: LD50 > 5000 mg/kg, male and female Fischer 344 rats, OECD 401, EPA OPPTS 870.1100, EU Method B.1 and Japan MAFF Acute Oral Toxicity Study, Stebbins & Brooks 1999.

Inhalation toxicity: LD50 > 5.24 mg/L (aerosol dust), male and female Fischer 344 rats, OECD 403, EPA OPPTS 870.1300, EU Method B.3, and Japan MAFF testing guidelines, Clements 2000.

Dermal toxicity: LD50 > 5000 mg/kg, male and female New Zealand White rabbits, OECD 402, EPA OPPTS 870.1200, EU Method B.3 and Japan MAFF Acute Dermal Toxicity Study, Stebbins & Brooks 1999.

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:

9 August 1999 to 24 August 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Acute Oral Toxicity Study, 1985.

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 106-165 g
- Fasting period before study: rats were fasted the night prior to treatment.
- Diet: pelleted rodent diet, ad libitum.
- Water: municipal water, ad libitum.
- Acclimation period: at least two weeks prior to study start.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 40-70 %
- Air changes (per hr): 12-15 times/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/dark photocycle, lights on at 6:00 a.m. and off at 6:00 p.m.

IN-LIFE DATES: From: 14 June 1999 To: 24 August 1999

Route of administration:

oral: gavage

Vehicle:

other: 0.5 % methylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 50 %

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

Five per sex per dose.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Detailed clinical observations were performed each day during the study.
Hand-held and open-field observations included a careful physical examination were performed during the study.
Each animal was weighed prestudy, the day of treatment, and on days 2, 8, and 15.
- Necropsy of survivors performed: yes
All rats submitted alive for necropsy were anesthetised by inhalation of methoxyflurane vapours and euthanised by decapitation after exposure and clamping of the trachea. A complete necropsy was conducted on all animals by a veterinary pathologist assisted by a team of trained individuals. The eyes were examined in situ using a moistened glass microscope slide applied to the corneal surface. Following inspection of the externum and body orifices, the nasal, cranial, oral, thoracic, and abdominal cavities were opened and the visceral organs were examined both in situ and following dissection.

Statistics:

Means and standard deviations were calculated for body weights. The data were evaluated for statistical outliers by a sequential test (Grubbs, 1969), however, outliers were not routinely excluded from statistical analysis.

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: One male and one female had perineal faecal soiling on test days 1 or 2, and one male had perineal faecal soiling on test day 15. All other animals were clinically normal throughout the study.

Gross pathology:

There were no treatment related gross pathologic observations for any animal.

Table 1: Individual Body Weights

Dose (mg/kg)	Animal Sex	Animal No.	Day of Test
			-1	1	2	8	15
5000	Male	3901	172.6	160.5	179.1	202.2	225.6
		3902	175.9	164.5	181.8	207.4	231.9
		3903	170.5	157.5	175.3	198.0	219.9
		3904	176.4	164.2	182.7	205.0	232.4
		3905	167.4	154.1	172.4	196.3	216.8
		Mean	172.6	160.2	178.3	201.8	225.3
		SD	3.8	4.4	4.4	4.6	7.0
	Female	3906	120.7#	111.2#	123.1	140.9	151.9
		3907	115.1	106.3	120.5	132.1	145.0
		3908	116.3	106.0	121.2	135.0	146.3
		3909	115.9	105.9	120.7	134.2	144.3
		3910	115.4	105.8	120.6	133.2	142.6
		Mean	116.7	107.0	121.2	135.1	146.0
		SD	2.3	2.3	1.1	3.4	3.5

# statistical outlier included

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the acute oral LD50 of the test material in male and female Fischer 344 rats was greater than 5000 mg/kg.

Executive summary:

The acute toxicity of the test material was determined in an acute oral toxicity test performed under GLP conditions and in accordance with the standardised guidelines OECD 401, EPA OPPTS 870.1100, EU Method B.1 and Japan MAFF Acute Oral Toxicity Study.

Five male and five female Fischer 344 rats received 5000 mg/kg of the test material as a 50 % mixture in 0.5 % methylcellulose by single dose oral gavage. Parameters evaluated during the two-week observation period included body weights and detailed clinical observations. All animals were examined for gross pathologic changes.

All animals survived for the duration of the study. One male and one female had perineal faecal soiling on test days 1 or 2, and one male had perineal faecal soiling on test day 15. All other animals were clinically normal throughout the study. Rats had a transient body weight loss on day 1, which was associated with the overnight fast prior to treatment, and then gained body weight for the remainder of the study. There were no treatment-related gross pathologic observations for any animal.

Under the conditions of the test, the acute oral LD50 of the test material in male and female Fischer 344 rats was greater than 5000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The key study was conducted under GLP conditions and followed standardised guidelines, producing reliable and conclusive results. The study was assigned a reliability score of 1, reliable without restrictions, in line with the principles of Klimisch et al (1997). A supporting study is also available. This was performed in accordance with generally accepted scientific principles, with incomplete reporting that did not affect the quality of the relevant results. This study was assigned a reliability score of 2, reliable with restrictions. The overall quality of the database is considered to be high.

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:

3 May 2000 (study initiation date)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guidelines

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: approximately 9 weeks of age (when exposed to the test material)
- Housing: Animals were housed in groups of 2-3 in stainless steel cages during acclimatisation. Animals were housed individually during exposure and the observation period.
- Diet: Pelleted rodent diet, ad libitum.
- Water: Water from the municipal supply, ad libitum.
- Acclimation period: Rats were acclimatised to test conditions at least 7 days prior to study initiation, and to nose cones for a single 2 hour period on the day preceding exposure.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Under controlled conditions.
- Humidity (%): Under controlled conditions.
- Photoperiod (hrs dark / hrs light): 12hr light/dark photoperiod.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Modified ADG nose-only chamber, 30 cm diameter x 60 cm height.
- Exposure chamber volume: 42 litres.
- Method of holding animals in test chamber: Animals were held in a chamber so that their nose was located in the exposure port.
- Source and rate of air: Compressed air (ambient temperature) was supplied at an approximate rate of 30 L/min, sufficient to provide 43 air changes per hour.
- System of generating particulates/aerosols: Dust aerosol of the test material was generated using a Jet Mill. A cyclone was placed between the dust generator and the chamber to further reduce the particle size of the aerosol in the chamber.
- Method of particle size determination: The aerodynamic particle size was determined twice during the exposure period through a six-stage cascade impactor by drawing samples, at a set rate, using a Sierra series 110 constant flow air sampler, through a vertical stainless steel tube which projected into the animal breathing zone. The mass median aerodynamic diameter and geometric standard deviation were determined for each sample as well as the average of both samples.
Samples were obtained at 65 and 195 minutes into the exposure time, at a sample rae of 3 L/min for 1 minute.
- Treatment of exhaust air: Air exiting the exposure chamber was passed through an absolute filter before the air was exhausted. The test material was not recycled.
- Temperature, humidity, pressure in air chamber: exposure was performed at room temperature, chamber temperature, humidity and airflow were recorded approximately every 30 minutes during the exposure period.
Mean values:
Chamber temperature: 19.5 ± 0.10 °C
Exposure room temperature: 19.7 ± 0.12 °C
Relative humidity: 40.3 ± 0.5 %
Chamber airflow: 30 L/minute

TEST ATMOSPHERE
- Brief description of analytical method used: The mass concentration of aerosol present in the chamber was determined gravimetrically five times during the exposure period. Samples were taken by drawing air, at a set rate, through a vertical stainless steel tube that projected into the animal breathing zone. Aerosol particles were collected on pre-weight 0.45 mircron TefSep, Teflon laminated 47 mm filters. The time-weighted average (TWA) exposure concentration was calculated from the gravimetric measurements.
Nominal concentrations were calculated based on the amount of test material fed into the generation system and the total chamber airflow.
- Samples taken from breathing zone: Yes.

TEST ATMOSPHERE
- Particle size distribution: Approximately 9 % of the total mass of particles were less than 1 micron in size and approximately 84 % were less than 6 microns in size.
- MMAD (Mass median aerodynamic diameter): The average MMAD of the aerosol particles was 3.86 microns (based on two determinations), with geometric standard deviation of 1.62. MMAD ranged from 1 to 4 µm.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

determined gravimetrically

Duration of exposure:

4 h

Concentrations:

TWA 5.24 mg/L, nominal concentration 11.40 mg/L.

No. of animals per sex per dose:

Five per sex per dose.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Animals were weighted and examined prior to exposure. Animals were observed at least every 30 minutes during the exposure period and each working day during the post exposure observation period.
Observations included an evaluation of the fur, eyes, mucous membranes and respiration.
Behaviour pattern and nervous system activity were assessed by specific observation for tremors, convulsions, salivation, lacrimation and diarrhoea, lethargy and other signs of altered central nervous system function.
An additional daily observation and routine monitoring on weekends (and holidays) was limited to husbandry procedures required to ensure the availability of feed and water.
All surviving rats were weighed on test days 2, 4, 8, 11 and 15 during the post exposure observation period.
Detailed clinical observations were conducted on all animals once prior to the start of treatment and daily thereafter. This was conducted at approximately the same time each day and included cage-side, hand-held and open-field observations that were recorded categorically or using a explicitly defined scales.
- Necropsy of survivors performed: Yes, all rats were submitted for a complete gross necropsy examination on test day 15.
Necropsy included examination of the eyes with a microscope slide using fluorescent illumination. Tissues were not saved, and no histopathologic examination was performed.

Statistics:

Mean and standard deviations were calculated for descriptive purposes for chamber concentration (mean only), animal room humidity, animal body weights, exposure room and chamber temperature, humidity and airflow. The range of the animal room temperature was reported.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.24 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occured during the study.

Clinical signs:

other: The only clinical effect noted during the exposure was soiled furcoat (faecal soiling). Following exposure (test day 1), treatment related clinical observations included perineal soiling (urine and/or faecal), extensive body soiling (urine and faecal), or

Body weight:

Mean body weight losses of approximately 5 % and 4 % were noted on test day 2 in male and females, respectively. The body weight of the rats exceeded the pre-exposure mean value by test day 8.

Gross pathology:

There were no visible lesions attributable to exposure noted in any rats exposed to the test material at the test day 15-scheduled necropsy.

Table 1: Individual Body Weights

Dose (mg/L)	Animal Sex	Animal No.	Day of Test
			1	2	4	8	11	15
5.24	Male	2243	208.7	195.2	204.2	214.1	219.4	230.6
		2244	207.9	199.0	207.6	219.5	227.2	235.5
		2245	208.5	200.4	207.2	216.4	225.9	240.1
		2246	213.1	200.6	208.7	222.6	232.9	245.4
		2247	217.6	208.5	215.3	226.1	236.9	248.6
		Mean	211.2	200.7	208.6	219.7	228.5	240.0
		SD	4.2	4.8	4.1	4.8	6.7	7.3
	Female	2248	132.8	128.3	131.4	138.8	142.9	148.1
		2249	133.6	128.8	132.5	136.6	140.3	145.4
		2250	134.9	129.4	137.1	141.2	145.3	150.1
		2251	137.8	132.3	135.1	139.9	142.7	148.0
		2252	140.7	125.2	140.6	147.8	152.1	155.5
		Mean	136.0	130.8	135.3	140.9	144.7	149.4
		SD	3.3	2.9	3.7	4.2	4.5	3.8

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the acute inhalation LC50 of the aerosolized test material in male and female Fischer 344 rats was greater than 5.24 mg/L air.

Executive summary:

The acute inhalation toxicity of the test material was determined in an aerosol inhalation study performed under GLP conditions and in accordance with the standardised guidelines OECD 403, EPA OPPTS 870.1300, EU Method B.3, and Japan MAFF testing guidelines.

Five male and five female Fischer 344 rats were nose-only exposed to a concentration of 5.24 mg/L. The aerosol particle size distribution mass median aerodynamic diameter (MMAD) averaged 3.86 microns with an average geometric standard deviation of 1.62 microns. Approximately 9 % of the mean mass particles were less than 1 micron. Approximately 84 % of the mean mass particles were less than 6 microns.

All animals survived the 4-hour exposure as well as the 2-week post-exposure period. Treatment-related in-life effects observed following exposure included perineal soiling (urine and/or faecal), abdominal soiling (faecal), or extensive body soiling (urine and faecal). All animals appeared normal by test day 3. The mean body weights, for the male and female rats, were decreased by approximately 4 and 5 % on test day 2 respectively, and had exceeded pre-exposure mean values by test day 8. There were no visible lesions attributable to exposure noted in any rats exposed to the test material at the test day 15-scheduled necropsy.

Under the conditions of the test, the 4 hour acute inhalation LC50 of the aerosolised test material in male and female Fischer 344 rats was greater than 5.24 mg/L air.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The key study was conducted under GLP conditions and followed standardised guidelines, producing reliable and conclusive results. The study was assigned a reliability score of 1, reliable without restrictions, in line with the principles of Klimisch et al (1997). The quality of the database is considered to be high.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

female

Type of coverage:

occlusive

Vehicle:

not specified

Details on dermal exposure:

TEST SITE
- Area of exposure: The trunk (back) of each rabbit.
- Site preparation: The site was clipped free of hair 24 hours prior to dosing.
- Type of wrap if used: An occlusive wrap, covered with an elastic rabbit jacket.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test material was wiped off.
- Time after start of exposure: 24 hours after application.

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

2 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Observations: Animals were observed for mortality, signs of toxicity and dermal irritation. Bodyweight was recorded throughout the observations period.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Both animals survived the test period.

Clinical signs:

other: No clinical signs of systemic toxicity were observed.

Other findings:

Local skin reaction
Both rabbits had erythema on test day 2. The erythema was resolved in each rabbit by test day 3.

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the LD50 for female New Zealand White rabbits was greater than 2000 mg/kg.

Executive summary:

The acute dermal toxicity of the test material was determined in a study which followed a protocol similar to OECD 404.

A single application of 2000 mg/kg of the test material was applied for 24 hours under an occluded wrap to the backs of two female New Zealand White rabbits. The occluded wrap was covered with an elastic rabbit jacket. Residual test material was wiped off when the bandage was removed 24 hours after application.

Both animals survived the test period. No clinical signs of systemic toxicity were observed. Both rabbits had erythema on test day 2. The erythema was resolved in each rabbit by test day 3. Body weights were not significantly affected. Therefore, under the conditions of this study, the estimated acute dermal LD50 for female rabbits was greater than 2000 mg/kg.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 August 1999 (study initiation date)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Acute Dermal Toxicity Study, 1985

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.3-2.5 kg
- Diet: 5 ounces of pelleted rodent feed per day.
- Water: Municipal water was provided ad libitum.
- Acclimation period: Animals were acclimated to the laboratory environment for at least two weeks prior to study start.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 ± 3 °C
- Humidity (%):40 - 60 %
- Air changes (per hr): approximately 12-15 times/hour
- Photoperiod (hrs dark / hrs light): 12- hour light/dark photocycle, lights on at 6:00 a.m. and off at 6:00 p.m.

IN-LIFE DATES: From: 1 May 1999 To: 31 August 1999

Type of coverage:

occlusive

Vehicle:

other: 0.5 % methylcellulose

Details on dermal exposure:

TEST SITE
- Area of exposure: Trunk of each rabbit.
- Site preparation: Clipped free of fur the day prior to application of the test material.
- % coverage: Approximately 10 % of the surface area of the rabbits.
- Type of wrap if used: The site was covered with a gauze/cotton patch (approximately 10 x 14 cm), held in place by an elastic jacket.

REMOVAL OF TEST SUBSTANCE
- Washing: The skin was wiped thoroughly with a water-moistened, soft disposable towel and dried with a soft disposable towel.
- Time after start of exposure: 24 hours.

VEHICLE
- Amount applied: The test material was moistened with 12.0 mL of 0.5 % methylcellulose.

Duration of exposure:

24 hours

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

Five per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
A detailed clinical observation (DCO) was conducted for all rabbits prior to test material administration for comparison with the observations recorded throughout the study. Animals were observed a minimum of 2 times, one of which was a DCO, on the day of treatment. A DCO was done each day (including weekends and holidays) during the study. Hand-held and open-field observations included a careful physical examination.
For scored DCO’s, only observations other than normal were recorded. Observations were dictionary based, and the dictionary contained most of the common physical and neurologic abnormalities seen in toxicity studies. Since not all potential observations were contained in the dictionary, free-field descriptions also were allowed.
The rabbits were weighed pre-study, the day of treatment (day 1) and on test days 2, 8, and 15.
- Necropsy of survivors performed: Yes
At study termination, all animals were euthanized with Beuthanasia solution. A complete necropsy was conducted on all animals by a veterinary pathologist assisted by a team of trained individuals. The eyes were examined in situ using a moistened glass microscope slide applied to the corneal surface. Following inspection of the externum, including the treatment site and body orifices, the nasal, cranial, oral, thoracic, and abdominal cavities were opened and the visceral organs were examined both in situ and following dissection.

Statistics:

Means and standard deviations were calculated for body weights. The data were evaluated for statistical outliers by a sequential test (Grubbs, 1969), however, outliers were not routinely excluded from statistical analysis.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured during the study.

Clinical signs:

other: There were no clinical signs of systemic toxicity. All rabbits had reddened and thickened skin at the dermal test site on day 2. In addition, one male had perineal faecal soiling on day 2 only. In all animals, the reddening and thickening of the skin reso

Gross pathology:

There were no treatment-related gross pathologic observations.

Table 1: Individual Body Weights

Dose (mg/kg)	Animal Sex	Animal No.	Day of Test
			-4	1	2	8	15
5000	Male	3921	2368.0	2396.0	2305.6	2593.8	2621.1
		3922	2381.9	2402.7	2344.5	2665.3	2693.7
		3923	2354.8	2386.6	2341.8	2540.9	2542.6
		3924	2441.7	2507.3#	2406.6	2760.5	2737.1
		3925	2354.1	2401.1	2285.8	2621.9	2586.1
		Mean	2380.1	2418.7	2336.9	2636.5	2636.1
		SD	36.3	49.9	46.2	82.7	79.0
	Female	3926	2409.0	2336.1	2415.3	2516.7	2625.0
		3927	2257.0	2255.6	2078.7	2392.8	2449.1
		3928	2406.0	2400.9	2302.4	2575.0	2605.6
		3929	2382.4	2381.7	2270.2	2670.1	2542.9
		3930	2333.1	2334.3	2284.8	2481.0	2508.1
		Mean	2357.5	2341.7	2270.3	2527.1	2546.1
		SD	63.9	56.1	121.4	103.7	71.8

# statistical outliers inculded

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the acute dermal LD50 of the test material for male and female New Zealand White rabbits was greater than 5000 mg/kg.

Executive summary:

The acute dermal toxicity of the test material was determined in a study performed under GLP conditions and in line with the standardised guidelines OECD 402, EPA OPPTS 870.1200, EU Method B.3 and Japan MAFF Acute Dermal Toxicity Study, 1985.

Five male and five female New Zealand White rabbits received a single 24 hour exposure to 5000 mg/kg body weight applied to shaved intact skin. The test material was moistened with 12.0 mL of 0.5 % methylcellulose. Parameters evaluated included body weights, detailed clinical observations, and gross pathologic evaluations.

All rabbits survived the 5000 mg/kg dose level. There were no clinical signs of systemic toxicity. All rabbits had reddened and thickened skin at the dermal test site on day 2. In addition, one male had perineal faecal soiling on day 2 only. In all animals, the reddening and thickening of the skin resolved by day 4. All rabbits lost body weight on days 1 or 2, and then surpassed their pre-study weights by study termination. There were no treatment-related gross pathologic observations.

Under the conditions of the test, the acute dermal LD50 of the test material for male and female New Zealand White rabbits was greater than 5000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The key study was conducted under GLP conditions and followed standardised guidelines, producing reliable and conclusive results. The study was assigned a reliability score of 1, reliable without restrictions, in line with the principles of Klimisch et al (1997). A supporting study is also available. This was performed in accordance with generally accepted scientific principles, with incomplete reporting that did not affect the quality of the relevant results. This study was assigned a reliability score of 2, reliable with restrictions. The overall quality of the database is considered to be high.

Additional information

Oral Toxicity

In the key study (Stebbins & Brooks, 1999) the acute toxicity of the test material was determined in an acute oral toxicity test performed under GLP conditions and in accordance with the standardised guidelines OECD 401, EPA OPPTS 870.1100, EU Method B.1 and Japan MAFF Acute Oral Toxicity Study.

Five male and five female Fischer 344 rats received 5000 mg/kg of the test material as a 50 % mixture in 0.5 % methylcellulose by single dose oral gavage. Parameters evaluated during the two-week observation period included body weights and detailed clinical observations. All animals were examined for gross pathologic changes.

All animals survived for the duration of the study. One male and one female had perineal faecal soiling on test days 1 or 2, and one male had perineal faecal soiling on test day 15. All other animals were clinically normal throughout the study. Rats had a transient body weight loss on day 1, which was associated with the overnight fast prior to treatment, and then gained body weight for the remainder of the study. There were no treatment-related gross pathologic observations for any animal.

Under the conditions of the test, the acute oral LD50 of the test material in male and female Fischer 344 rats was greater than 5000 mg/kg.

 

This result is supported by the findings reported in the second study (Brooks, 1997), where the acute oral LD50 was also reported to be > 2000 mg/kg in a study that was performed in line with good scientific principles.

Inhalation Toxicity

In the key study (Clements, 2000) the acute inhalation toxicity of the test material was determined in an aerosol inhalation study performed under GLP conditions and in accordance with the standardised guidelines OECD 403, EPA OPPTS 870.1300, EU Method B.3, and Japan MAFF testing guidelines.

Five male and five female Fischer 344 rats were nose-only exposed to a concentration of 5.24 mg/L. The aerosol particle size distribution mass median aerodynamic diameter (MMAD) averaged 3.86 microns with an average geometric standard deviation of 1.62 microns. Approximately 9 % of the mean mass particles were less than 1 micron. Approximately 84 % of the mean mass particles were less than 6 microns.

All animals survived the 4-hour exposure as well as the 2-week post-exposure period. Treatment-related in-life effects observed following exposure included perineal soiling (urine and/or faecal), abdominal soiling (faecal), or extensive body soiling (urine and faecal). All animals appeared normal by test day 3. The mean body weights, for the male and female rats, were decreased by approximately 4 and 5 % on test day 2 respectively, and had exceeded pre-exposure mean values by test day 8. There were no visible lesions attributable to exposure noted in any rats exposed to the test material at the test day 15-scheduled necropsy.

Under the conditions of the test, the 4 hour acute inhalation LC50 of the aerosolised test material in male and female Fischer 344 rats was greater than 5.24 mg/L air.

Dermal Toxicity

In the key study (Stebbins & Brooks, 1999) the acute dermal toxicity of the test material was determined in a study performed under GLP conditions and in line with the standardised guidelines OECD 402, EPA OPPTS 870.1200, EU Method B.3 and Japan MAFF Acute Dermal Toxicity Study, 1985.

Five male and five female New Zealand White rabbits received a single 24 hour exposure to 5000 mg/kg body weight applied to shaved intact skin. The test material was moistened with 12.0 mL of 0.5 % methylcellulose. Parameters evaluated included body weights, detailed clinical observations, and gross pathologic evaluations.

All rabbits survived the 5000 mg/kg dose level. There were no clinical signs of systemic toxicity. All rabbits had reddened and thickened skin at the dermal test site on day 2. In addition, one male had perineal faecal soiling on day 2 only. In all animals, the reddening and thickening of the skin resolved by day 4. All rabbits lost body weight on days 1 or 2, and then surpassed their pre-study weights by study termination. There were no treatment-related gross pathologic observations.

Under the conditions of the test, the acute dermal LD50 of the test material for male and female New Zealand White rabbits was greater than 5000 mg/kg.

 

This result is supported by the findings reported in the second study (Brooks, 1997), where the acute dermal LD50 was also reported to be > 2000 mg/kg in a study that was performed in line with good scientific principles.

Justification for selection of acute toxicity – oral endpoint

The key study was conducted under GLP conditions, in accordance with standardised guidelines and is fully reported.

Justification for selection of acute toxicity – inhalation endpoint

Only one study is available. The key study was conducted under GLP conditions, in accordance with standardised guidelines and is fully reported.

Justification for selection of acute toxicity – dermal endpoint

The key study was conducted under GLP conditions, in accordance with standardised guidelines and is fully reported.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I (Classification and Labelling Requirements for Hazardous Substances and Mixtures), Regulation (EC) No. 1272/2008 (CLP), the substance does not require classification with respect to acute toxicity via the oral, inhalation or dermal route.