2,6-dibromo-4-cyanophenyl heptanoate

Endpoint summary

Description of key information

Key, M-280105-01-1; oral (rat, US EPA 81-1, GLP): LD50 = 362.4 mg/kg bw (males), LD50 = 291.9 mg/kg bw (females), LD50 = 322.4 mg/kg bw (combined)

Key, M-280109-01-1; inhalation (rat, US EPA 81-3, GLP): LC50 = ca. 1.975 mg/L (males), LC50 = ca. 1.479 mg/L (females), LC50 = ca. 1.7172 mg/L (combined)

Key, M-226977-01-1; dermal (rabbit, US EPA 81-2, GLP): LD50 > 2020 mg/kg bw (males and females)

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:

19 Jul - 17 Aug 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted 1987

Deviations:

yes

Remarks:

No details regarding environmental conditions.

Qualifier:

according to guideline

Guideline:

EPA OPP 81-1 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc., Houston, Texas
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adults
- Weight at study initiation: 234 - 309 g (males), 175 - 198 g (females)
- Fasting period before study: for at least 16 hours prior to treatment
- Housing: 1 - 3 per cage (separated by sex) suspended, wire bottom, stainless steel cages
- Diet: Purina Formulab Chow #5008, ad libitum
- Water: tap water, ad libitum
- Acclimation period: At least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): not reported
- Humidity (%): not reported
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): not reported

IN-LIFE DATES: From: 19 Jul To: 17 Aug 1989

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The dose was administered by gavage using an appropriately sized syringe and stainless steel ball-tipped intubation needle. The animals were returned to their cages immediately after treatment.

Doses:

249, 500 and 750 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: three times on the day of treatment and at least once daily thereafter
- Frequency of weighing: just prior to treatment and on Days 7 and 14, or at the time of discovery after death
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes

Statistics:

None

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

362.4 mg/kg bw

Based on:

test mat.

95% CL:

193.2 - 679.8

Remarks on result:

other: Mortality occurred starting from the lowest tested dose level of 249 mg/kg bw.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

291.9 mg/kg bw

Based on:

test mat.

95% CL:

157.1 - 542.1

Remarks on result:

other: Mortality occurred starting from the lowest tested dose level of 249 mg/kg bw.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

322.4 mg/kg bw

Based on:

test mat.

95% CL:

219.5 - 473.3

Remarks on result:

other: both sexes combined

Mortality:

- 249 mg/kg bw: 2/5 males and 2/5 females died (males: both on Day 1, females: 1 in the first 6 h after treatment, 1 on Day 2)
- 500 mg/kg bw/day: 2/5 males and 4/5 females died (all deaths occurred during the first 6 h after treatment, except 1 male which died on Day 1)
- 750 mg/kg bw: 5/5 males and 5/5 females died (all males died within 3 h after treatment, all females on Day 1)

Clinical signs:

other: - 249 mg/kg bw: signs of salivation, lacrimation and nasal discharge, piloerection (reversible within 4 days), activity decrease (reversible within 1 day) - 500 mg/kg bw: signs of lacrimation, nasal discharge and salivation, signs of nasal discharge, pilo

Gross pathology:

A gross pathological examination was conducted on each animal which died during the study and on each animal which survived through termination of the study. Of the gross pathological findings noticed, those described as signs of lacrimation, nasal discharge, polyuria, and salivation; discoloration of the contents of the gastrointestinal tract, discoloration of the lungs, gastrointestinal tract distended with gas, large intestine empty, and variations thereof were considered possibly related to the treatment.

Summarized results can be found in Attachment 1.

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The study was conducted under GLP conditions, according to U.S. EPA guideline No. 81-1, which in principle is similar to the OECD test guideline 401. The resulting LD50 for male and female rats was 362 and 292 mg/kg bw, respectively; the combined LD50 was 322.4 mg/kg bw. Taking into account the most sensitive LD50 value of 292 mg/kg bw which refers to female rats, the substance is to be classified in Cat 3 according to the criteria of the the CLP Regulation (EU) No. 1272/2008.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

292 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable guideline study (Klimisch score 2), and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No 1907/2006.

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:

31 Aug - 21 Sep 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 2009

Deviations:

yes

Remarks:

Limited information on environmental conditions, exposure and test atmosphere generation.

Qualifier:

according to guideline

Guideline:

EPA OPP 81-3 (Acute inhalation toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc., Houston, Texas
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adult
- Weight at study initiation: 299 - 370 g (males), 176 - 240 g (females)
- Housing: in groups of 1-3, separated by sex, in suspended, wire bottom, stainless steel cages
- Diet: Purina Formulab Chow #5008, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): not reported
- Humidity (%): not reported
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): not reported

IN-LIFE DATES: From: 31 Aug To: 21 Sep 1989

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

not specified

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

> 1.651 - < 1.978 µm

Geometric standard deviation (GSD):

> 1.866 - < 2.279

Remark on MMAD/GSD:

Details can be found in Attachment 1.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 200 L New York University design, stainless steel, dynamic flow inhalation chamber
- Exposure chamber volume: 200 L
- Method of holding animals in test chamber: not specified
- Source and rate of air (airflow): 73.3 - 77.6 L/min
- System of generating particulates/aerosols: aerosol was generated by pumping the test material through a pressure operated spray nozzle. This was then diluted with dried and filtered air and drawn into the exposure chamber.
- Method of particle size determination: Particle size determinations were made using an Andersen cascade impactor.
- Temperature, humidity, pressure in air chamber: Temperature and humidity were recorded at 30 minute intervals during the exposure period from a Taylor wet bulb/dry bulb hygrometer located in the exposure chamber, temperature ranged from 22.8 - 25.5 °C, relative humidity from 59 - 75%, air pressure was not specified.

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: Analysis of the concentration of the test atmosphere was performed according to a modification of the method provided by the sponsor using a Tracer Model 560 gas chromatograph. The nominal concentration was determined by dividing the loss in weight of the test material after each exposure by the total volume of air which passed through the chamber.
- Time needed for equilibrium of exposure concentration before animal exposure: not reported
- Particle size distribution: Please refer to Attachment 1

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Nominal: 1.94 , 8.54 and 21.1 mg/L
Measured: 0.523, 1.45 and 2.69 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: on the day of exposure and at least once daily thereafter. Due to chamber design, only 4 animals (2 males and 2 females, for example) could be observed during the exposure period. Observations after four hours included all animals.
- Frequency of weighing: just prior to the inhalation exposure and on Days 7 and 14 or at the time of discovery after death
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes

Statistics:

None

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

1.975 mg/L air

Based on:

test mat.

95% CL:

1.433 - 2.722

Exp. duration:

4 h

Remarks on result:

other: Mortality occurred at the mid and high concentration levels of 1.45 and 2.69 mg/L air, respectively.

Key result

Sex:

female

Dose descriptor:

LC50

Effect level:

1.479 mg/L air

Based on:

test mat.

95% CL:

0.748 - 2.924

Exp. duration:

4 h

Remarks on result:

other: Mortality occurred at the mid and high concentration levels of 1.45 and 2.69 mg/L air, respectively.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

1.717 mg/L air

Based on:

test mat.

95% CL:

1.267 - 2.327

Exp. duration:

4 h

Remarks on result:

other: both sexes combined

Mortality:

- 0.523 mg/L: no mortality observed
- 1.45 mg/L: 1/5 males died, 3/5 females died
- 2.91 mg/L: 4/5 males died, 4/5 females died

Clinical signs:

other: clinical signs of toxicity in males and females almost included nasal discharge, salivation, lacrimation, piloerection, ptosis, and decreased activity; for details, please refer to "any other Information on results incl. tables", and to Attachment 2.

Body weight:

- 0.523 mg/L: all animals achieved body weight gains as expected during the 14 day observation period.
- 1.45 mg/L: surviving animals achieved body weight gains as expected over the 14 day observation period despite of a lost in weight noticed during the first week.
- 2.69 mg/L: the 2 surviving animals achieved body weight gains as expected over the 14 day observation period despite of a lost in weight noticed during the first week..

Summarized results can be found in Attachment 3.

Gross pathology:

- 0.523 mg/L: No pathological findings were reported.
- 1.45 mg/L: lungs mottled red (1 male and 3 females); orange and red mucoid material in small intestine (1 male and 2 females)
- 2.69 mg/L: lungs red or dark red (3 males, 3 females); orange mucoid material in small intestine (2 males, 2 females), tan slurry and dark brown slurry in stomach (2 males), gastrointestinal tract empty (1 female), dark red mucoid material in small intestine (1 female)

Summarized results can be found in Attachment 3.

Other findings:

None

Details on clinical signs of toxicity:

- 0.523 mg/L: clinical signs of toxicity for both, males and females, consisted of ptosis, decrease in activity and piloerection; the signs were all reversible.
- 1.45 mg/L: nasal discharge, salivation, lacrimation, piloerection, decrease in activity and ptosis were the more common signs of toxicity seen in males and females at this concentration level.
- 2.69 mg/L: also at this concentration level, the clinical signs of toxicity almost consisted of nasal discharge, salivation, lacrimation, piloerection, decrease in activity and ptosis for both, males and females.
Please also refer to Attachment 2.

 

 

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The study was conducted under GLP conditions, according to U.S. EPA guidelines No. 81-3, which in principle is similar to the OECD test guideline 403. The animals were exposed for 4 hours to the test item as aerosol. The resulting combined LC50 for male and female rats was set at 1.72 mg/L air. Accordingly, the substance is to be classified in Cat 4 according to the criteria of the the CLP Regulation (EU) No. 1272/2008.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

1.72 mg/L air

Physical form:

inhalation: aerosol

Quality of whole database:

The available information comprises an adequate and reliable guideline study (Klimisch score 2), and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No 1907/2006.

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:

13 Jul - 27 Jul 1989

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: Fixed Dose Procedure)

Version / remarks:

adopted 2017

Deviations:

yes

Remarks:

No details regarding environmental conditions.

Qualifier:

according to guideline

Guideline:

EPA OPP 81-2 (Acute Dermal Toxicity)

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
- Source: Ray Nichols Rabbitry, Lumberton. Texas
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adults
- Weight at study initiation: 2.725 - 3.300 kg (males), 2.400 - 3.425 (females)
- Fasting period before study: no
- Housing: individually in suspended, wire bottom, stainless steel cages
- Diet: Purina Rabbit Chow; ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): not reported
- Humidity (%): not reported
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): not reported

IN-LIFE DATES: From: 12 Jul To: 27 Jul 1989

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: at least 10
- Type of wrap if used: Surgical gauze (10 x 10 cm and two layers thick) was applied to the trunk of the animal and held in place with non-irritating adhesive tape. Then, the trunk was wrapped with a semi-occlusive dressing held in place non-irritating adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing: with room temperature tap water and a clean wet cloth
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 1.32 mL/kg bw
- Constant volume or concentration used: yes
- For solids, paste formed: no but heated to 45 °C in a water bath to liquefy the test substance

Duration of exposure:

24 h

Doses:

2020 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: 1/2, 3 and 6 h after treatment and at least once daily thereafter
- Frequency of weighing: on Days 0, 7 and 14 or at the time of discovery after death
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 020 mg/kg bw

Based on:

test mat.

Remarks on result:

other: except for one female, no mortality occurred.

Mortality:

1 female died on Day 2. The gross necropsy examination conducted on this animal and on each animal which survived through termination of the study revealed no obvious abnormalities.

For details, please refer to Attachment 1.

Clinical signs:

other: - decreased defecation (1 female, Day 2 - 4, 3 males on Day 2 and 3, 1 male on Day 4) - diarrhea (1 female, Day 13) For details, please refer to Attachment 2.

Gross pathology:

At necropsy, the gross pathological examination conducted on the animal that died on Day 2 and on each animal which survived through termination of the study revealed no obvious abnormalities.

Interpretation of results:

GHS criteria not met

Conclusions:

The study was conducted under GLP conditions, according to U.S. EPA guideline No. 81-2, which in principle is similar to the OECD test guideline 402. The resulting LD50 was > 2020 mg/kg bw for both, male and female rabbits. According to criteria of the CLP Regulation (EU) No. 1272/2008, no classification of the test item for acute dermal toxicity is warranted.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 020 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable guidleine study (Klimisch score 2), and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No 1907/2006.

Additional information

To assess the acute toxicity of the test substance, the reliable data as available for each of the oral, inhalation and dermal route of exposure are considered.

Acute oral toxicity

The acute oral toxicity of the test substance was determined according to the US EPA guideline 81-1, which in principle is similar to the OECD test guideline 401, and in compliance with GLP (M-280105-01-1). Groups of five male and female rats were administered single oral doses of 249, 500 and or 750 mg/kg bw. No animals of the high dose group survived the treatment. In the group that received 249 mg/kg bw of the test substance, 2/5 males and 2/5 females died, in the 500 mg/kg bw group, 2/5 males and 4/5 females died. All deaths occurred between 3 h after treatment and the afternoon of Day 2. Treatment related clinical signs included e.g. lacrimation, nasal discharge, polyuria, and salivation. All surviving animals gained body weight as expected during the observation period. At necropsy, discoloration of the contents of the gastrointestinal tract, discoloration of the lungs, gastrointestinal tract distended with gas and empty large intestines were observed. Based on the mortality observed during the study, the LD50 value (acute median lethal oral dose) of 362.4 mg/kg bw for males and 291.9 mg/kg bw for females was derived. The combined LD50 was set at 322.4 mg/kg bw.

 

Acute toxicity: inhalation

A GLP-conform acute inhalation toxicity study was performed according to the US EPA guideline 81-3, which in principle is similar to the OECD test guideline 403 (M-280109-01-1). Five Sprague Dawley rats/sex/dose were exposed to the test item (aerosole) for 4 h. The test concentrations comprised 0.523, 1.45, and 2.69 mg/L air (analytical concentration). The mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) corresponding to the dosage groups were (MMAD / GSD): 1.64 µm / 2.27 µm (0.523 mg/L), 1.96 µm / 1.97 µm (1.45 mg/L) and 1.83 µm / 1.92 µm (2.69 mg/L). Post exposure, all animals were intended to be observed for a period of 14 days. In the high dose group, 4 male and 4 female animals died and in the middle dose group, 1 male and 3 female rats died within the 14 day observation period. In contrast, all animals of the 0.523 mg/L air group survived until scheduled sacrifice.

All animals of the low concentration group achieved body weight gains as expected during the 14 day observation period. However, clinical signs were observed such as ptosis, decrease in activity and piloerection. All of these were reversible within 1 day after treatment. In the middle and high concentration group, surviving animals achieved body weight gains over the 14 day observation period , however with some lost of weight during the first week. In animals of both groups activity decrease, lacrimation, nasal discharge, piloerection, ptosis, and salivation were observed. In animals of the middle and high dose group that survived the 14 day observation period, no pathological abnormalities were found at necropsy. Animals that died before the end of the study showed discoloration of the contents of the gastrointestinal tract, an empty gastrointestinal tract and discoloration of the lungs (red to dark red).

The LC50 value derived from this study was ca. 1. 1.975 mg/L air for male rats and 1.479 mg/L air for female rats. Thus, the resulting combined LC50 for the test item as aerosol was 1.72 mg/L air with respect to male and female rats.

 

Acute dermal toxicty

A GLP-conform acute dermal toxicity study was performed according to the US EPA guideline 81-2, which in principle is similar to the OECD test guideline 402 (M-226977-01-1). A limit dose of 2020 mg/kg bw was topically applied for 24 h to the clipped skin of five male and female rabbits under semi-occlusive dressing. On Day 2 after exposure, one female died. One other female and three male animals had decreased defecation on Days 2-4 and in one female, diarrhea was observed on Day 13. Most rabbits achieved body weight gains as expected during both weeks of the 14 day observation period with the exception of 1 female and 1 male. No gross abnormalities were observed at necropsy. Thus, the LD50 value in male and female rabbits was greater than 2020 mg/kg bw.

Justification for classification or non-classification

According to criteria of the CLP Regulation (EU) No. 1272/2008, classification of the test item for acute oral toxicity category 3 and acute inhalation toxicity category 4 is warranted. No classification is needed with respect to acute dermal toxicity.

The substance is listed in Annex VI of Regulation (EC) 1272/2008 (607-427-00-7). Harmonised classification in regard to minimum classification was adopted according to the available experimental data if appropriate.