Reaction mass of 2-[[[2-[(2-aminoethyl)amino]ethyl]amino]carbonyl]-benzoic acid and 2,2' -[iminobis(2,1-ethanediyliminocarbonyl)]bis-benzoic acid and 7,8,9,10,11,12,20,21,22,23,24,25-dodecahydrodibenzo[i,t] [1,4,7,12,15,18] hexaazacyclodocosine-5,13,18,26(6H,19H)-tetrone

Endpoint summary

Description of key information

One acute oral study, one acute inhalation study and one acute dermal study were performed with the test substance.

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:

1973

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP study.

Qualifier:

no guideline available

Principles of method if other than guideline:

In this acute oral range finding study, 6 males albino rats were used to performed the expriment. 2 rats for each dose levels were used (1000 / 3000 / 10000 mg/kg). Clinical signs, mortality and body weight were recorded.

GLP compliance:

no

Test type:

other: No data

Limit test:

no

Species:

rat

Strain:

other: Albino rats

Sex:

male

Details on test animals or test system and environmental conditions:

No data

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

Form administred: 25.0% (w/v) suspension in corn oil.

Doses:

1000 / 3000 and 10000 mg/kg

No. of animals per sex per dose:

2 males for each dose level.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At test day 0 and test day 14 for the surviving
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.

Statistics:

No data

Sex:

male

Dose descriptor:

LD50

Effect level:

> 1 000 - < 3 000 mg/kg bw

Based on:

test mat.

Mortality:

- At 1000 mg/kg: 0 percent dead.
- At 3000 mg/kg: 100 percent death after 30 minutes after administration.
- At 10000 mg/kg: 100 percent death (the first one 30 minutes after administartion and 15 minutes after administration for the second)

Clinical signs:

other: - At 1000 mg/kg: Hypoactivity, gasping, pilo-erection. - At 3000 mg/kg: Hypoactivity, gasping, abdominal griping, pilo-erection, convulsions. - At 10000 mg/kg: Hypoactivity, gasping, abdominal griping, pilo-erection, convulsions.

Gross pathology:

Gross pathology finding: Necropsy examination of the animals that died revealed hemorrhaged stomachs. No gross pathologic alterations were noted among the animals sacrified at the end of the 14-day observation period.

Reactions:

Dose (mg/kg)	Reaction	Time of onset following dose administration	Duration of reaction	Time of death following dose administration
1 000	Hypoactivity	5 minutes	6 -22 hours	/
1 000	Gasping	5 minutes	10 minutes	/
1 000	Pilo-erection	5 minutes	6 -22 hours	/
3 000	Hypoactivity	5 minutes	Until death	30 minutes
3 000	Gasping	5 minutes	Until death	30 minutes
3 000	Abdominal griping	5 minutes	Until death	30 minutes
3 000	Pilo-erection	15 minutes	Until death	30 minutes
3 000	Convulsions	15 minutes	Until death	30 minutes
10 000	Hypoactivity	5 minutes	Until death	15 -30 minutes
10 000	Gasping	5 minutes	Until death	15 -30 minutes
10 000	Abdominal griping	5 minutes	Until death	15 -30 minutes
10 000	Pilo-erection	15 minutes	Until death	15 -30 minutes
10 000	Convulsions	15 minutes	Until death	15 -30 minutes

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In the acute oral toxicity experiment, the LD50 was determined to be > 1000 mg/kg <3000 mg/kg.

Executive summary:

In the acute oral range finding study, 6 males albino rats were used.

2 rats for each dose levels were used (1000 / 3000 / 10000 mg/kg).

The test material was suspended in corn oil before administration to the rats.

Mortality

- At 1000 mg/kg: 0 percent dead.

- At 3000 mg/kg: 100 percent death after 30 minutes after administration.

- At 10000 mg/kg: 100 percent death (the first one 30 minutes after administartion and 15 minutes after administration for the second)

Clinical signs:

- At 1000 mg/kg: Hypoactivity, gasping, pilo-erection.

- At 3000 mg/kg: Hypoactivity, gasping, abdominal griping, pilo-erection, convulsions.

- At 10000 mg/kg: Hypoactivity, gasping, abdominal griping, pilo-erection, convulsions.

In conclusion, in the acute oral toxicity experiment, the LD50 was determined to be > 1000 mg/kg <3000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

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:

1973

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP study.

Qualifier:

no guideline available

Principles of method if other than guideline:

This acute dust inhalation toxicity study was conducted wherein a group of ten albino rats was exposed to a dust of the test item for four hours in a 70 -liter inhalation chamber. The average analytical concentration was 1.05 mg/L air. After exposure, the test animals were observed for 14 days.

GLP compliance:

no

Test type:

other: not available

Limit test:

yes

Species:

rat

Strain:

other: Albino rats

Sex:

not specified

Details on test animals or test system and environmental conditions:

10 albino rats were used.

Route of administration:

inhalation: dust

Type of inhalation exposure:

whole body

Vehicle:

not specified

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION

- Exposure chamber volume: 70-liter inhalation chamber

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

4 h

Concentrations:

1.05 mg/L air

No. of animals per sex per dose:

10 albino rats

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

not specified

Dose descriptor:

LC50

Effect level:

> 1.05 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no death during the inhalation exposure or observation period.

Clinical signs:

other: Untoward bahavioral reactions exhibited by the rats included hypoactivity, lacrimation and conjunctivitis.

Body weight:

Body weight gains of nine out of ten rats were normal.

Gross pathology:

Necropsy examinations, performed on all rats at the termination of the two weeks observation period revealed two test rats with tan nodules on all lobes of the lungs. All other finding in the test animals were essentially the same as in stock animals taken from the same group as the test rats.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

LC50 is greater than 1.05 mg/L air in rats.

Executive summary:

An acute dust inhalation toxicity study was conducted wherein a group of ten albino rats was exposed to a dust of the test material for four hours in a 70 -liter inhalation chamber. The avrage analytical concentration was 1.05 mg/L air. After exposure, the test animals were observed for 14 days.

Untoward behavioral reactions exhibited by the test animals during exposure included lacrimation, hypoactivity and conjunctivitis. No deaths were noted during the exposure or the two-week observation period. Body weight gains were normal except for one rat.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

1 050 mg/m³ air

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:

1973

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP study. The experiment was performed according to the guideline equivalent or similar to the OECD 402 (Acute dermal toxicity).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

not specified

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

other: Albinos rabbits

Sex:

male/female

Details on test animals or test system and environmental conditions:

6 albinos rabbits (3males and 3 females)

Type of coverage:

not specified

Vehicle:

other: Test material was applied to premoistened skin.

Duration of exposure:

24 hours

Doses:

300/ 1000 / 3000 mg/g

No. of animals per sex per dose:

Two animals for each dose level (one male and one female)

Control animals:

not specified

Details on study design:

Test item was applied at each dose level on one abraded and one intact animal.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 3 000 other: mg/kg

Based on:

test mat.

Mortality:

No mortality

Clinical signs:

other: The rabbits did not exhibit any unusual behavioral reactions following dermal exposure. Local skin recation after 24-hour exposure period were characterized by barely perceptible pale to red erythema. No skin reaction were noted at 7 and 14 days.

Gross pathology:

Necropsy examination of all rabbits sacrified at the end of the 14-day observation period, did not reveal any gross pathologix alterations.

Mortality and body weight:

Dose* (mg/kg)	Animal N° and sex	Individual bodyweight (kg) at test day 0	Individual bodyweight (kg) at test day 14	Number dead/ Number tested	Percent dead
300	1 -M** 2 -F	2.86 2.64	2.70 2.98	0/2	0
1000	3-M** 4 -F	2.90 2.44	2.80 2.56	0/2	0
3000	5-M** 6 -F	2.98 3.18	2.96 3.14	0/2	0

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute LD50 of the test material was determined to be greater than 3000 mg/kg.

Executive summary:

Three males and three females albinos rabbits were used to detemined the acute dermale toxicity of the test material.

Two animals for each dose level (one male and one female): 300 / 1000 and 3000 mg/kg.

Test item was applied at each dose level on one abraded and one intact animal.

The observation period is 14 days after 24 hours exposure duration.

The rabbits did not exhibit any unusual behavioral reactions following dermal exposure. Local skin recation after 24-hour exposure period were characterized by barely perceptible pale to red erythema. No skin reaction were noted at 7 and 14 days.

Necropsy examination of all rabbits sacrified at the end of the 14-day observation period, did not reveal any gross pathologix alterations.

No mortality occured during the observation time.

The acute LD50 of the test material was dezermined to be greater than 3000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

The acute oral toxicity study was performed in male rats and the test substance was applied at a dose level of 1000, 3000 and 10000 mg/kg. All animals died at the dose level of 3000 and 10000 mg/kg whereas no death occurred at the dose level of 1000 mg/kg. The Oral LD50 was therefore considered to be >1000 < 3000 mg/kg.

An acute inhalation study was performed with the test substance. The test substance was applied during 4 -hours to male rats at a measured average concentration of 1.05 mg/L air. No death occured during the study and following observation period. The Acute Inhalation LC50 is greater than 1.05 mg/L air.

An acute dermal toxicity was conducted with albino rabbits at a dose level of 300, 1000 and 3000 mg/kg. The substance was applied to the intact skin during 24 hours and no death occured during the study and following observation period. The acute dermal LD50 in rabbit is considered to be greater than 3000 mg/kg.

Justification for selection of acute toxicity – oral endpoint
only existing study

Justification for selection of acute toxicity – inhalation endpoint
Only existing study

Justification for selection of acute toxicity – dermal endpoint
Only existing study

Justification for classification or non-classification

- oral toxicity:

Based on the above stated assessment of the acute oral toxicity the substance does need to be classified as Acute Oral toxicity Category 4 according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and accordingCLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council)as implementation of UN-GHS in the EU.

- dermal toxicity:

Based on the above stated assessment of the acute dermal toxicity of the substance (absence of toxicity up to 3000 mg/kg) the substance does not need to be classified according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and accordingCLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council)as implementation of UN-GHS in the EU.

- inhalation toxicity:

Based on the above stated assessment of the acute inhalation toxicity of the substance (absence of toxicity up to 1.05 mg/L) the substance does not need to be classified according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and accordingCLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council)as implementation of UN-GHS in the EU.