N,N'-[6,13-diacetamido-2,9-diethoxy-3,10-triphenodioxazinediyl]bis(benzamide)

Endpoint summary

Description of key information

Several acute oral toxicity studies were performed, resulting in LD50 values of >5000 and >15000 mg/kg bw. In an acute dermal toxicity study (OECD guideline 402, GLP) no mortality occured and no signs of toxicity were observed. LD50 dermal is considered to be > 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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CERJ., 53940 Le Genest
- Weight at study initiation: 220-250 g (males), 170-200 g (females)
- Fasting period before study: 18 hours
- Housing : 10 animals per Makrolon SAFI 60/40 cm cage, with dust free white wood shavings
- Diet: UAR A04, 18 to 25 g (depending on age and weight of the animal)
- Water: tap water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21
- Humidity (%): 40-60
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25 g/100 mL

MAXIMUM DOSE VOLUME APPLIED: 2.0 mL/100 g

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

- Duration of observation period following administration: animals were observed continuously on the first day after administration, they then remained in daily observation for 14 days
- Necropsy performed: yes, animals that died during the first day (8 hours) were dissected and their major organs (liver, spleen, kidneys, stomach, lung, heart) examined macroscopically. All organs with obvious macroscopic lesions were retained for further histological examination if necessary.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

other: Immediately after administration of Violet Cromophtal B, animals are lethargic and their fur is ruffled.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

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:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Purity: 98.3%

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 243 - 203g
- Housing: singly
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5d
- Fasting: 16h before administration

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

corn oil

Remarks:

suspension

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal and dorso-lateral parts of the trunk
- % coverage: 10
- Type of wrap if used: semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 6.67 g/kg bw (paste)
- For solids, paste formed: yes

Duration of exposure:

24h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: days 0, 7, 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

Calculations were performed using Microsoft Excel 2003 and checked with a calculator.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality occurred.

Clinical signs:

other: No signs of systemic toxicity were observed in the animals.

Gross pathology:

No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study.

Other findings:

Violet discoloration of the skin at the application site (evaluation of erythema formation was not possible)

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the median lethal dose (LD50) of the test substance after dermal application was found to be greater than 2000 mg/kg bw in male and female rats.

Executive summary:

In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the test substance (as suspension/paste in corn oil Ph.Eur.) to the clipped skin (dorsal and dorso-lateral parts of the trunk) and covered by semi-occlusive dressing for 24 hours. The application area comprised at least 10% of the total body surface area. The animals were observed for 14 days.

No mortality occurred.

No signs of systemic toxicity were observed in the animals.

The following test item-related local effects were recorded during the course of the study, local effects occurred within the first 3 days after administration:

- Violet discoloration of the skin at the application site (evaluation of erythema formation was not possible)

- No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study.

The mean body weight of the male animals increased within the normal range throughout the study period. In the female group 4 animals showed a stagnation or marginal loss of body weight during the first week, but body weights were in a normal range during the second week.

This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth. Due to the fact that stagnation of body weight is commonly known for females dermally applied, this stagnation is considered to be unspecific. Accordingly, the acute dermal median lethal dose (LD50) was determined to be greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

In an acute oral toxicity study male and female Tif RAI rats were administered the test substance at dose levels of 1000, 3000, 10000 and 15000 mg/kg bw (5 animals per sex per dose group) by oral gavage. Sesame oil was used as vehicle. Dosing was followed by a 15 day observation period. At 1000 and 3000 mg/kg bw no clinical signs were observed. At 10000 and 15000 reduction in spontaneous motility, diarrhea and violett colored skin were observed. No mortality was reported. Therefore the LD50 of the test substance is > 15000 mg/kg bw.

In another acute oral toxicity study male and female Wistar rats were administered the test substance at a dose levels of 5000 mg/kg bw (10 animals per dose group) by oral gavage. Arachis oil was used as vehicle. Dosing was followed by a 14 day observation period. Immediately after dosing, animals are lethargic and their fur is ruffled. No mortality was reported. Therefore the LD50 of the test substance is > 5000 mg/kg bw.

In an acute dermal toxicity study (Limit Test), young adult Wistar rats were dermally exposed to a single dose of 2000 mg/kg bw of the test item to the clipped skin and covered by semi-occlusive dressing for 24 hours. No mortality occurred. No signs of systemic toxicity were observed in the animals. Violet discoloration of the skin at the application site (evaluation of erythema formation was not possible) was observed. No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study. The mean body weight of the male animals increased within the normal range throughout the study period. In the female group 4 animals showed a stagnation or marginal loss of body weight during the first week, but body weights were in a normal range during the second week. This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth. Due to the fact that stagnation of body weight is commonly known for females dermally applied, this stagnation is considered to be unspecific. Accordingly, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw.

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.