Benzonitrile, 3,3'-(2,3,5,6-tetrahydro-3,6-dioxopyrrolo[3,4-c]pyrrole-1,4-diyl)bis-

Endpoint summary

Description of key information

A single dose of the test item was administered orally or dermally to Wistar rats at concentrations of 5000 and 2000 mg/kg bw, respectively. No mortality occurred. Clinical examiniation and gross necropsy did not reveal any findings. The LD50 after oral administration is therefore considered to be > 5000 mg/kg bw and after dermal application > 2000 mg/kg bw.

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:

1992-06-24 to 1993-03-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant Guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted Sep. 19, 1984

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Appearance: orange powder

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Olac Ltd., Bicester, Oxon, England
- Age at study initiation: four to seven weeks
- Weight at study initiation: 110 to 120 g
- Fasting period before study: yes
- Housing: in groups of five according to sex in metal cages with wire mesh floors
- Diet: standard laboratory rodent diet (Biosure LAD 1) ad libitum
- Water: drinking water ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 to 22°C
- Humidity: 49% (daily mean)
- Air changes: approximately 10 - 15 per hour
- Photoperiod: 12 hours artificial light (0700 - 1900 hours) in each 24-hour period

IN-LIFE DATES: From: 1993-01-19 To: 1993-02-04

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
A 25% w/v suspension of test item was prepared on the day of dosing in 1 % w/v aqueous methylcellulose.

VEHICLE
- Concentration in vehicle: 1 %
- Amount of vehicle: 20 mL / kg bw / day

Doses:

2.5 g / kg bw (preliminary study)
5 g / kg bw (main study)

No. of animals per sex per dose:

2 males and 2 females (preliminary study)
5 males and 5 females (main study)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Cage side observations: at least twice daily
- Clincal signs: soon after dosing and at frequent intervals for a period of 7 hours. On subsequent days once in the morning and at the end of the experimental day.
- Body weight: on Days 1 (prior to dosing), 8 and 15

- Necropsy of survivors performed: yes

Statistics:

No

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths following a single oral dose of test item.

Clinical signs:

other: Pilo-erection was observed in all rats within four minutes of dosing and throughout the remainder of Day 1. Recovery of all rats, as judged by external appearance and behaviour, was complete by Day 2. Orange/brown faeces were observed on Day 2 only.

Gross pathology:

No macroscopic abnormalities were observed for animals killed on Day 15.

Results of preliminary study:

The acute lethal oral dose of the test item to male and female rats was greater than 2.5 g/kg bodyweight.

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

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:

1992-06-24 to 1993-04-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant Guideline study.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted Sep. 19, 1984

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Appearance: orange powder

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Olac Ltd., Bicester, Oxon, England
- Age at study initiation: seven to ten weeks
- Weight at study initiation: 207 to 258 g
- Fasting period before study: no
- Housing: individually in metal cages with wire mesh floors
- Diet: standard laboratory rodent diet (Biosure LAD 1) ad libitum
- Water: drinking water ad libitum
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 to 22°C
- Humidity: 49% (daily mean)
- Air changes: approximately 10 to 15 per hour
- Photoperiod: 12 hours artificial light (0700 - 1900 hours) in each 24-hour period

IN-LIFE DATES: From: 1993-01-19 To: 1993-02-02

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

distilled

Details on dermal exposure:

TEST SITE
- Area of exposure: dorso-lumbar region
- % coverage: approximately 10% of the total body surface
- Type of wrap if used: the gauze, which covered the application site, was held in place with a non-irritative dressing

REMOVAL OF TEST SUBSTANCE
- Washing: the treated area of skin was washed with warm (30° to 40°C) water and blotted dry with absorbent paper
- Time after start of exposure: at the end of the 24 hours exposure period

TEST MATERIAL
- Amount applied: 2.0 g / kg bodyweight
- Administration volume: 4.0 mL / kg bodyweight
- Concentration: 50 % w/v in distilled water

Duration of exposure:

24 hours

Doses:

2.0 g / kg bodyweight

No. of animals per sex per dose:

5 males and 5 females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Cage side observations: at least twice daily
- Clincal signs: soon after dosing and at frequent intervals for a period of 5 hours. On subsequent days once in the morning and at the end of the experimental day.
- Body weight: on Days 1 (prior to dosing), 8 and 15

- Necropsy of survivors performed: yes
- Dermal responses: erythema, eschar and oedema formation (daily scoring, 0 - 4)

Statistics:

No

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths following a single dermal application of test item at 2.0 g / kg bodyweight.

Clinical signs:

other: There were no signs of systemic reaction to treatment.

Gross pathology:

No macroscopic abnormalities were observed for animals killed on Day 15.

Other findings:

DERMAL RESPONSES
Staining (orange) prevented assessment of erythema for all rats on Day 2; no evidence of oedema was recorded at this time. Although some residual (orange) staining was apparent on Days 3 and 4, there were no signs of erythema or oedema at any of the application sites from Day 3 to the end of the study.

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Additional information

A single dose of the test item was administered by gavage to male and female Sprague-Dawley rats at concentrations of 5000 mg/kg bw. The animals were observed for 14 days. Mortality, body weight and clinical signs were recorded continuously. Organ weighing and gross necropsy was performed after scheduled sacrifice.

A single dose of the test item was applied onto skin of male and female Sprague-Dawley rats at concentrations of 2000 mg/kg bw. The animals were observed for 14 days. Mortality, body weight and clinical signs were recorded continuously. Organ weighing and gross necropsy was performed after scheduled sacrifice.

No mortality occurred, body weight and body weight gain were comparable to control animals. Clinical signs of toxicity were not observed and gross necropsy did not reveal any findings. Orange/brown faeces were observed on Day 2 after oral application. The LD50 is therefore considered to be > 5000 (oral) and > 2000 mg/kg bw (dermal), respectively.

Justification for classification or non-classification

                      Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for acute toxicity under Regulation (EC) No. 1272/2008.