N-ethyl-4-[[4-(ethylamino)-m-tolylphenyl][4-(ethylimino)-3-methylcyclohexa-2,5-dien-1-ylidene]methyl]-m-toluidine monoacetate

Endpoint summary

Description of key information

Based on the findings with the acute toxic class method, an LD50 of > 350 and < 1400 mg/kg bw was determined for male and female Wistar rats (recalculated based on dye content in registered substance (74% w/w) versus tested material (52% w/w)).

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:

2000-05-18 to 2000-07-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and guideline compliant study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

(1996)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

(1996)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

(1998)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Deutschland, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: male animals approx. 8 - 12 weeks, female animals approx. 13 - 18 weeks
- Weight at study initiation: males: 178 - 183 g; females: 171 g
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: Single housing
- Diet: ad libitum (Kliba-Labordiaet, Provimi Kliba SA, Kaiseraugst, Switzerland)
- Water: ad libitum (tap water)
- Acclimation period: for at least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 5 and 20 g/100 mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: Aqueous formulation corresponds to the physiological medium.

CLASS METHOD
- Rationale for the selection of the starting dose: Based on the physical and chemical characteristics of the test substance and its composition, some acute oral toxicity was expected. Therefore, a starting dose of 500 mg/kg body weight has been chosen in the first step with 3 male animals. As none of those animals died, 2000 mg/kg body weight were administered to 3 male animals in a second step. Because all animals died at the second step, 500 mg/kg body weight have been tested in a third step with 3 female animals. Because 2 animals survived the third step the study was terminated.

Doses:

200, 500 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 17 days
- Frequency of observations and weighing: observations were done at least once each workday; weighing was performed at the start of the study, weekly thereafter and at the end of the study
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 500 - < 2 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 350 - < 1 400 mg/kg bw

Based on:

act. ingr.

Remarks on result:

other: recalculated based on dye content of registered substance (74% w/w) versus tested material (52% w/w)

Mortality:

All animals of. the 2000 mg/kg dose group died until study day 8. One female animal of the 500 mg/kg dose group died on study day 2.

Clinical signs:

other: Male animals: Signs of toxicity noted in the 2000 mg/kg dose group comprised impaired and poor general state, dyspnoea, gasping, apathy, abdominal position, staggering, paresis, tremor, piloerection, smeared fur, diarrhea, reduced feces, exsiccosis, viole

Gross pathology:

Necropsy findings of the animal that died at 500 mg/kg comprised moderate dilation of stomach, small intestine and large intestine as well as blue and/or red discoloration of the stomach content. In the animals that died at 2000 mg/kg blue and or red discoloration of intestinal tract content and same organs and tissues was observed. No macroscopic pathologic abnormalities were noted in the surviving animals examined at the end of the observation period.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

350 mg/kg bw

Quality of whole database:

Acceptable and well documented study.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In an acute oral toxicity study according to OECD guideline 423 and EU method B.1 tris, single doses of 2000 and 500 mg/kg body weight of test material preparations in doubly distilled water were given to 3 test groups of fasted animals (2000 mg/kg in 3 males, 500 mg/kg in 3 males and 3 females) by gavage in a sequential manner (BASF AG, 2001).

All animals of the 2000 mg/kg dose group died until study day 8. One female animal of the 500 mg/kg dose group died on study day 2. Signs of toxicity noted in the 2000 mg/kg dose group comprised impaired and poor general state, dyspnoea, gasping, apathy, abdominal position, staggering, paresis, tremor, piloerection, smeared fur, diarrhea, reduced feces, exsiccosis, violet discoloured feces and urine. Findings were observed until study day 8.

Signs of toxicity noted in the 500 mg/kg dose group comprised impaired and poor general state, red clammy snout, dyspnoea, gasping, squatting posture, apathy, staggering, tremor, piloerection, smeared fur, red brown discoloured urine. Findings were observed until study day 17.

The mean body weights of the male animals of the 500 mg/kg test group decreased during the first post exposure observation week but increased during the second week.

The mean body weights of the surviving female animals of the 500 mg/kg test group decreased during the first two post exposure observation weeks but increased during the third week.

Necropsy findings of the animal that died at 500 mg/kg comprised moderate dilation of stomach, small intestine and large intestine as well as blue and/or red discoloration of the stomach content. In the animals that died at 2000 mg/kg blue and or red discoloration of intestinal tract content and some organs and tissues was observed.

No macroscopic pathologic abnormalities were noted in the surviving animals examined at the end of the observation period. Under the conditions of this study the median lethal dose of the test substance after oral administration was found to be greater than 500 mg/kg and less than 2000 mg/kg body weight for male and female rats based on the active ingredient. With regard to the actual test item composition the median lethal dose was recalculated to greater than 350 mg/kg and less than 1400 mg/kg body weight.

Justification for selection of acute toxicity – oral endpoint

GLP and guideline compliant study report

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance needs to be classified and labelled as acute tox. cat. 4, H302 "Harmful if swallowed." under Regulation (EC) No 1272/2008, as amended for the sixth time in Directive EC 605/2014.