1,4-bis(mesitylamino)anthraquinone

Endpoint summary

Description of key information

Acute oral toxicity: 
The test item did not cause any mortality, signs of intoxication, or necropsy findings after single oral gavage administration to male and female rats at 5000 mg/kg bw in a GLP compliant guideline study. Thus the LD50 value of the test item was found to be greater than 5000 mg/kg bw.
Acute dermal toxicity: 
The test item did not cause any mortality, signs of intoxication, or necropsy findings after single dermal administration to male and female rats at 2000 mg/kg bw in a GLP compliant guideline study. Thus the LD50 value of the test item was found to be greater than 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:

1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well performed GLP compliant OECD guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Principles of method if other than guideline:

The limit dose applied was 5000 mg/kg bw.

Allthough the wording of study title "RANGE-FINDING" purports a reduced study design a full limit test was performed according to OECD guidline 401.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bantin & Kingman Ltd., Grimston, Aldborough, Hull, U.K.
- Age at study initiation: approximately five to eight weeks
- Weight at study initiation: males : 126 - 138 g, females: 120 - 125 g,
- Fasting period before study: one night before dosing, 2 hours after dosing
- Housing: in groups of up to five by sex in solid-floor polypropylene cages with sawdust bedding
- Diet: Rat and Mouse Expanded Diet No.1, Special Diet Services Limited, Witham, Essex, U.K (ad libitum):
- Water: drinking water (ad libitum):
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 45 - 64
- Air changes (per hr): approx. 15
- Photoperiod : 12 hrs dark / 12 hrs light

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

Concentration: 500 mg/ml
Dose volume: 10 ml/kg bw

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

five males and five females

Control animals:

no

Details on study design:

All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.

Deaths and overt signs of toxicity were recorded 1/2, I, 2 and 4 hours after dosing and subsequently once daily for 14 days.
Individual bodyweights were recorded on the day of treatment (day 0) and on days 7 and 14.

Preliminary study:

DOSE LEVEL CONCENTRATION DOSE VOLUME NUMBER OF RATS
mg/kg mg/ml ml/kg MALE FEMALE
5000 500 10 1 1
2000 200 10 1 1
200 20 10 1 1

No deaths occured during the observation time of 5 days

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths

Clinical signs:

other: No signs of systemic toxicity

Gross pathology:

No abnormalities were noted at necropsy of animals killed at the end of the study

Other findings:

No other findings reported

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral median lethal dose (LD5O) of the test material in the Sprague-Dawley strain rat was found to be greater than 5000 mg/kg bodyweight.

Executive summary:

A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 401 "Acute Oral Toxicity" referenced as Method B1 in Commission Directive 84/449/EEC.

Following a range-finding study (1 male, 1 female with the doses 5000, 2000 and 200 mg/kg bw), a group of ten fasted animals (five males and five females) was given a single oral dose of test material preparation, administered as a solution in arachis oil B.P. at a dose level of 5000 mg/kg bodyweight.

There were no deaths after a observation period of 14 days. All animals showed expected gain in bodyweight during the study. Neither signs of systemic toxicity nor abnormalities were noted.

Therefore, the acute oral median lethal dose (LD5O) of the test material in the Sprague-Dawley strain rat was found to be greater than 5000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Quality of whole database:

reliable without restrictions - well performed GLP compliant OECD guideline study

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:

June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well performed GLP compliant OECD guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A., Calco (Lecco), Italy
- Age at study initiation: 7-9 weeks
- Weight at study initiation: Approximately 200-240 g
- Housing: Individually caged (both during acclimatisation and study period)
- Diet (e.g. ad libitum): 4 RF 18 (Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI) Italy); Ad libitum throughout the study
- Water (e.g. ad libitum): Drinking tap water supplied to each cage via a water bottle; Ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 2°C
- Humidity (%): 55% ± 15%
- Air changes (per hr): Approximately 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours

IN-LIFE DATES:
From: 13 June 2012 – Allocation of the animals
To: 28 June 2012 – Necropsy of the animals

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

A volume of 1 mL of sterile water was added to weighed aliquot of the test item for each animal and mixed in order to achived a paste for the dosing procedure

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal surfaces of the trunk of the animals
- % coverage: approximately 10% of body surface
- Type of wrap if used: A strip of synthetic film was placed over the treated site and the whole assembly held in place by encircling the trunk of the animal with a length of elastic adhesive bandage, this forming a semi-occlusive barrier.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treatment area was cleaned by gentle swabbing of the skin with cotton wool soaked with lukewarm water.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Aliquots were weighed according to the body weight of each animal measured prior to dosing
- For solids, paste formed: yes

Duration of exposure:

Single treatment

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days after dosing
- Frequency of observations and weighing:
Mortality and morbidity twice daily
Weighed on allocation, Days 1, 8 and 15 of the study period
Clinical signs on the day of dosing: At dosing, approximately 1, 2 and 4 hours after dosing, daily thereafter (14 days).
- Necropsy of performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality at 2000 mg/kg bw, the highest dose tested

Mortality:

No mortality occurred in male or female animals following treatment.

Clinical signs:

other: Clinical signs recorded during the observation period were limited to the treatment site (dorsum), both in male and female animals, in which slight/marked blue coloration was generally observed during the entire period of the study. In addition, blue stai

Gross pathology:

No internal abnormalities were found at necropsy examination performed on all animals at termination of the study.
At the external examination, all animals showed blue staining at the treated site (dorsum) and on the tail surface.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The results obtained indicate that the test item has no systemic toxic effect in the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg bw. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg bw.

Executive summary:

The acute toxicity of the test item was investigated following dermal administration of a single dose to the rat.

A single dose of 2000 mg/kg bw moistened with water was administered to a group of 5 male and 5 female animals for 24 hours. After 14 days all animals were killed and subjected to necropsy examination.

No mortality occurred following treatment in male or female animals during the observation period.

Clinical signs recorded during the observation period were limited to blue coloration of the treated site and on the tail surface during the entire period of the study. This blue coloration was attributed to the properties of the test item (blue). Scab on the dorsum was observed in one female only from Day 7 to Day 11. This sign was considered incidental and not related to the tolerability of the test item.

The body weight changes observed during the study were within the expected range for this species and age of animals.

No internal abnormalities were found at necropsy carried out in the animals at termination of the study. External examination revealed blue staining at the treated site (dorsum) and on the tail surface in all animals.

These results indicate that the test item has no systemic toxic effect in the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg bw. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

reliable without restrictions - well performed GLP compliant OECD guideline study

Additional information

Justification for selection of acute toxicity – oral endpoint
no alternative study available

Justification for selection of acute toxicity – dermal endpoint
no alternative study available

Justification for classification or non-classification

LD50 values derived from acute dermal and oral toxicity studies were greater than 2000 mg/kg bw. Additionally no specific target organ toxicity was observed. Since these findings do not meet the criteria for classification with respect to acute toxicity or specific target organ toxicity (STOT SE) according to the rules laid down in Directive 67/548EEC and in Regulation (EC) No 1272/2008, classification is not warrantable.