5-bromo-1,3-dichloro-2-fluorobenzene

Endpoint summary

Description of key information

Oral LD50 (rat): 2000 mg/kg bw < LD50 < 5000 mg/kg bw (GLP, OECD TG 425)

Dermal LD50 (rat) >5000 mg/kg bw (GLP, OECD TG 402)

Inhalation LC50 (rat) >5.36 mg/L (GLP, OECD TG 403)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Remarks:

The study was conducted to meet the national regulatory requirements in a non-EEA country.

Type of information:

experimental study

Adequacy of study:

key study

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:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

up-and-down procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SAGE® Labs
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 164-210 grams
- Fasting period before study: overnight
- Housing: Animals were group housed, except on the day of administration, at which time they were single housed and until the animals were deemed acceptable, based on observations, to return to group housing
- Diet (e.g. ad libitum): Envigo Teklad Global 16% Protein Rodent Diet #2016 ad libitum (except for pre-dose fast)
- Water (e.g. ad libitum): Filtered tap water ad libitum
- Acclimation period: 6-25 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 51-77
- Air changes (per hr): 12 or 13
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

IN-LIFE DATES: Start: June 5, 2018; End: July 19, 2018

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

In an acute oral toxicity study, a group of fasted,
8-12 weeks old, female, Sprague-Dawley derived, albino rats were given a single oral dose of CA5528. Prior to dosing, the rats were fasted overnight. During the fasting period, the rats were examined for health and weighed. Individual doses were calculated based on the initial body weights, taking into account the density of the test substance.

Doses:

5000 mg/kg or 2000 mg/kg

No. of animals per sex per dose:

Four at 5000 mg/kg; five at 2000 mg/kg

Control animals:

no

Details on study design:

The animals were observed for mortality, signs of gross toxicity, and behavioural changes within the first several hours post-dosing and at least once daily thereafter for up to 14 days after dosing or until death occurred. 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, diarrhea and coma.
Individual body weights of the animals were recorded prior to test substance administration (initial day 0) and again on days 7 and 14 (termination) following dosing or after death.

One rat was euthanized for humane reasons due to test substance exposure. Surviving rats were euthanized on Day 14. All animals were euthanized via CO2 inhalation. Gross necropsies were performed on all deceased and euthanized animals. Tissues and organs of the thoracic and abdominal cavities were examined.

Statistics:

Statistical analysis was limited to the calculation of the mean density value for dosing.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 - < 5 000 mg/kg bw

Based on:

test mat.

Mortality:

Three of four animals receiving 5000 mg/kg died, whereas there was no mortality at 2000 mg/kg.

Clinical signs:

other: 5000 mg/kg dose level (4 animals): Two females died within six days of test substance administration. One animal was euthanized for humane reasons on Day 3. Prior to death, all animals were hypoactive and exhibited irregular respiration, nasal discharge

Gross pathology:

5000 mg/kg dose level (4 animals): Gross necropsy of the deceased animals revealed distention of the intestines and/or stomach. No gross abnormalities were noted for the animal euthanized for humane reasons on Day 3. No gross abnormalities were noted for the surviving animal when necropsied at the conclusion of the 14-day observation period.
2000 mg/kg dose level (5 animals): No gross abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the conditions of this study, the acute oral median lethal dose (LD50) of CA5528 is between 2000 mg/kg and 5000 mg/kg of body weight in female rats.

Executive summary:

The acute oral toxicity test was conducted with rats to determine the potential for CA5528 - (Composition: CA5528, 99.8% w/w) to produce toxicity from a single dose via the oral route.

An initial limit dose of 5000 mg/kg of body weight was administered to one healthy female rat by oral gavage.  Due to the absence of mortality in this animal, two additional females received the same dose level, simultaneously.  Since both animals died, one additional animal was tested. Due to the mortality in this animal, the study proceeded to a limit test at 2000 mg/kg of body weight at the request of the Sponsor.  An initial limit dose of 2000 mg/kg was administered to one healthy rat by oral gavage.  Due to the absence of mortality in this animal, four additional females received the same dose level, sequentially.  Since these animals survived, no additional animals were tested.  Females were selected for the test because they are frequently more sensitive to the toxicity of test compounds than males.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30-60 minutes, 1 and approximately 3 hours after treatment on day 1 and once daily during test days 2 to 14 or until death.  Body weights were recorded on day 1 (prior to test substance administration) and on days 7 and 14 or after death.  All animals were examined macroscopically.

Of the 4 animals administered a limit dose of 5000 mg/kg, two females died within six days of test substance administration.  One animal was euthanized for humane reasons on Day 3.  Prior to death, all animals were hypoactive and exhibited irregular respiration, nasal discharge, reduced fecal volume, ocular discharge, piloerection, soft feces, prone posture and/or was moribund. One animal survived the limit dose of 5000 mg/kg.  Following administration, the surviving animal was hypoactive and exhibited irregular respiration, abnormal gait, facial staining, ocular discharge, hunched posture, abdominal distention and ano-genital staining.  However, the animal recovered by Day 6 and appeared active and for the remainder of the 14-day observation period, gaining body weight over the course of the study.  Gross necropsy of the deceased animals revealed distention of the intestines and/or stomach.

At 2000 mg/kg dose level (5 animals), all animals survived exposure to the test substance and gained body weight.  Following administration, two animals were hypoactive and four animals exhibited irregular respiration, facial staining, reduced fecal volume, oral discharge ano-genital staining and/or were cold to the touch.  However, these animals recovered by Day 6 and appeared active and healthy for the remainder of the study. One animal did not exhibit any signs of toxicity.

The body weight of the surviving animals was within the range commonly recorded for this strain and age.

No macroscopic findings were recorded for the survivors at necropsy.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Quality of whole database:

A fully reliable and valid study according to relevant OECD test guidelines and GLP is available.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Remarks:

The study was conducted to meet the national regulatory requirements in a non-EEA country.

Type of information:

experimental study

Adequacy of study:

key study

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SAGE® Labs USA
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10-11 weeks
- Weight at study initiation: 332-396 g (males), 210-240 g (females)
- Fasting period before study: no
- Housing: The animals were housed in caging which conforms to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals (Natl. Res. Council, 2011). Animals were group housed, except on the day of exposure, at which time they were singly housed and until the animals were deemed acceptable, based on observations, to return to group housing.
- Diet (e.g. ad libitum): Envigo Teklad Global 16% Protein Rodent Diet #2016 ad libitum (except during exposure)
- Water (e.g. ad libitum): Filtered tap water ad libitum (except during exposure)
- Acclimation period: 19 or 23 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 50-77
- Air changes (per hr): 12 or 13
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

IN-LIFE DATES: Start: June 8, 2018; End: July 9, 2018

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

3.68 µm

Geometric standard deviation (GSD):

2.4

Remark on MMAD/GSD:

MMAD was measured at 1.5 and 3 hours into exposure, and measured values were 3.94 and 3.41 µm, respectively. Geometric standard deviation was 2.44 and 2.33.

Details on inhalation exposure:

Exposure conditions: Prior to initiation of the full inhalation study, pre-test trials were conducted to establish generation procedures to achieve, to the extent possible, the desired chamber and desired particle size distribution (mass median aerodynamic diameter between
1 and 4 μm). Exposure conditions during the study are given later in a table of the test atmosphere characteristics of CA5528.
Generation of the test atmosphere/chamber description: A nose-only inhalation chamber was used for exposure. Animals were individually housed in polycarbonate holding tubes which seal to the chamber with an “O” ring during exposure. The base unit terminates the chamber with a 0.5-inch diameter tube for discharged air.
Air Supply: Filtered generator air was supplied to the spray atomization nozzle by an air compressor, and measured with a Mass Flow Controller. Additional filtered mixing air from the same air compressor, measured with a Mass Flow Controller, was introduced into the chamber to help uniformly distribute the test atmosphere by creating a vortex at the chamber inlet. Chamber airflow was monitored throughout the exposure period and recorded periodically. The exposure was conducted under slight negative pressure.
Ambient Conditions: The temperature and relative humidity within the exposure chamber as well as the room were monitored continuously during exposure, and were measured with a temperature-humidity monitor. Temperature and relative humidity values were recorded every 15 minutes for the first hour of exposure and approximately every 15 or 30 minutes thereafter.
Atmosphere Generation: The test atmosphere was generated using a nebulizer. The test substance was metered to the atomization nozzle through PharMed® BPT tubing, using a peristaltic pump.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Remarks on duration:

The animals were exposed to the targeted chamber concentration for at least 4 hours. The exposure period was extended beyond 4 hours to allow the chamber to reach equilibrium (T99).

Concentrations:

5.36 mg/L

No. of animals per sex per dose:

Five

Control animals:

no

Details on study design:

Fourteen healthy, naive rats not previously tested were selected for exposure. Two males and two females were selected for the sighting test at 5.0 mg/L and five males and five females were selected for the main test at 5.0 mg/L. Prior to the start of the study they 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 or until death. 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, diarrhea and coma. The body weight of each rat was recorded prior to exposure and on days 1, 3, 7 and 14.
All rats were euthanized via CO2 inhalation at the end of the 14-day observation period. Gross necropsies were performed on all animals. Tissues and organs of the thoracic and abdominal cavities were examined.

Statistics:

Statistical analysis was limited to the calculation of the mean and standard deviation. Since no death occurred at the limit dose, the LC50 was determined without the need of statistical analysis.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.36 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no mortalities at a concentration of 5.37 mg/L and there were no mortalities at a concentration of 5.36 mg/L.

Clinical signs:

other: All animals exhibited irregular respiration and/or ano-genital staining. However, all animals recovered by Day 3 and appeared active and healthy for the remainder of the study.

Body weight:

All animals gained body weight during the study.

Gross pathology:

Apart from discoloration of the lungs in one male and one female, there were no other gross abnormalities noted for any animal when necropsied at the conclusion of the 14-day observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the acute inhalation median lethal concentration (LC50) of CA5528 is greater than 5.36 mg/L in Sprague-Dawley derived, albino male and female rats.

Executive summary:

In an acute inhalation toxicity study, groups of young adult Sprague-Dawley-derived, albino rats (5/sex/group) were exposed via the inhalation (nose-only exposure) route to CA5528.  Test atmospheres were analyzed for particulate concentration.

After establishing the desired generation procedures during the pre-test trials, fourteen healthy rats were selected for test.  A sighting test with an initial exposure level of 5.0 mg/L was selected for testing using two animals per sex (2 males and 2 females).  Since all male and female rats survived the sighting test, an additional five animals per sex (5 males and 5 females) were selected for the main test with a 5.0 mg/L exposure level.  Chamber concentration and particle size distributions of the test atmosphere were determined periodically during the exposure period.  The animals were observed for mortality, signs of gross toxicity, and behavioral changes at least once daily for 14 days following exposure. Body weights were recorded prior to exposure (initial) and again on Days 1, 3, 7, and 14 (terminal). Necropsies were performed on all animals at terminal sacrifice.

The achieved test atmosphere had the following characteristics:

Sighting study (5 mg/L): 5.37 ± 0.29 mg/L achieved, MMAD: 2.97; 3.22 µm, GSD: 2.45; 2.45

Main study (5 mg/L): 5.36 ± 0.48 mg/L achevied, MMAD: 3.94; 3.41 µm, GSD: 2.44; 2.33

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

A fully reliable and valid study according to relevant OECD test guidelines and GLP is available.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Remarks:

The study was conducted to meet the national regulatory requirements in a non-EEA country.

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SAGE® Labs
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult (10-12 weeks)
- Weight at study initiation: males 334-376 grams and females 210-239 grams
- Fasting period before study: no
- Housing: Animals were group housed, except on the day of application, at which time they were singly housed until the animals were deemed acceptable, based on observations, to return to group housing.
- Diet (e.g. ad libitum): Envigo Teklad Global 16% Protein Rodent Diet #2016 ad libitum
- Water (e.g. ad libitum): Filtered tap water ad libitum
- Acclimation period: 21 or 29 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 52-77
- Air changes (per hr): 12 or 13
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

IN-LIFE DATES: Start: June 14, 2018; End: July 11, 2018

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Initial testing was conducted on one male and one female rat to determine extreme clinical signs or severe irritation potential of the test substance. Due to the absence of clinical signs or severe local signs of irritation, an additional four males and four females were tested. Five thousand milligrams of the test substance per kilogram of body weight was applied evenly over a dose area of approximately 2 inches x 3 inches (approximately 10% of the body surface) and covered with a 2-inch x 3-inch, 4-ply gauze pad. The gauze pad and entire trunk of each animal were then wrapped with 3-inch Durapore tape to avoid dislocation of the pad and to minimize loss of the test substance. The rats were then returned to their designated cages. The day of application was considered Day 0 of the study.
After 24 hours of exposure to the test substance, the pads were removed and the test sites were gently cleansed with a 3% soap solution followed by tap water and a paper towel to remove any residual test substance.

Duration of exposure:

24 hours

Doses:

5000 mg/kg bw (Individual doses were calculated based on the initial body weights, taking into account the density of the test substance)

No. of animals per sex per dose:

Five (initial test was done with one animal per sex)

Control animals:

no

Details on study design:

The animals were observed for mortality, signs of gross toxicity, and behavioural changes during the first several hours after application 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, diarrhea and coma.
Individual body weights of the animals were recorded prior to test substance application (initial) and again on day 7 and day 14 (terminal).
All surviving rats were killed by CO2 inhalation at the end of the 14-day observation period.
Gross necropsies were performed on all animals killed at the end of the study. Tissues and organs of the thoracic and abdominal cavities were examined.

Statistics:

Statistical analysis was limited to the calculation of the mean density value for dosing. Since no deaths occurred at the limit dose, the LD50 was determined without the need of 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

Clinical signs:

other: All animals survived test substance administration during the study. Other than the dermal irritation noted at the dose site of all animals between Days 1 and 8, there were no other adverse clinical findings recorded for any animal over the course of the

Gross pathology:

No gross abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the single dose acute dermal median lethal dose (LD50) of CA5528 is greater than 5000 mg/kg of body weight in male and female rats.

Executive summary:

An acute dermal toxicity test was conducted with rats to determine the potential for CA5528 to produce toxicity from a single topical application.  Initially, 5000 mg/kg of the test substance was applied to the skin of one healthy male rat and one healthy female rat for 24 hours. Due to the absence of clinical signs or severe local signs of irritation, an additional four males and four females were tested.  The animals were observed for mortality, signs of gross toxicity, and behavioral changes at least once daily for 14 days. Body weights were recorded prior to application (initial), on Day 7, and on Day 14 (terminal). Necropsies were performed on all animals.

All animals survived test substance administration and gained body weight during the study.  Other than the dermal irritation noted at the dose site of all animals between Days 1 and 8, there were no other adverse clinical findings recorded for any animal over the course of the study.  No gross abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

A fully reliable and valid study according to relevant OECD test guidelines and GLP is available.

Justification for classification or non-classification

The substance does not meet the criteria for classification for acute toxicity in accordance with CLP-Regulation (EC) No. 1272/2008.