1,4-bis(p-tolylamino)anthraquinone

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats for the given test chemical. The LD50 value is >5000 mg/kg bw. The study concluded that the LD50 value is >2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to very low vapour pressure of the test chemical, which is reported to be 9.40E-11 Pa (7.05E-013 mmHg). Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on different studies conducted on rats and rabbits for the given test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

data is from study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

Determination of the acute oral toxicity of the test substance in rats over a period of 14 observation days.

GLP compliance:

no

Remarks:

pre-dates GLP-regulation

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

- Identity: 1,4-bis(p-tolylamino)anthraquinone(BAG)
OTHER SPECIFICS: Safety precautions: Routine hygienic procedure was sufficient to assure personnel health and safety.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: KLEINTIERFARM MADOERIN AG.; 4414 FUELLINSDORF/SWITZERLAND
- Age at study initiation: approximately 9 weeks
- Weight at study initiation: Males: 150-212 g; Females: 151-191 g
- Fasting period before study: Overnight
- Housing: The animals were caged in groups of five in Macrolon cages type 3 with wire mesh tops and standardized granulated soft wood bedding.
- Identification: By cage number and individual colour spots.
- Diet (e.g. ad libitum): Pelleted standard Kliba 24/343/1 rat maintenance diet,ad libitum.
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-2 degC
- Humidity (%): 55 +/-10 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours/day

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 400

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 10-25 %
- Amount of vehicle (if gavage): 10-20 ml
- Justification for choice of vehicle: no data

MAXIMUM DOSE VOLUME APPLIED: 20 ml/kg

Doses:

Group 1: 1000 mg/kg bw
Group 2: 5000 mg/kg bw

No. of animals per sex per dose:

5 males , 5 females per group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Frequency of weighing: 1/week
- Necropsy of survivors performed: yes
- Other examinations performed:
- Mortality: Five times during the day and daily thereafter.
- Body weight: Body weights were recorded at the day of administration and days 7 and 14 after the administration.

Statistics:

no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed

Mortality:

Mortality was not observed.

Clinical signs:

other: 1000 mg/kg: Dyspnea, curved body position, diarrhoea and ruffeld fur. 5000 mg/kg: Sedation, dyspnea, exophthalmos, curved body position, diarrhoea and ruffeld fur. All animals had recovered within 7 observation days.

Gross pathology:

No macroscopical organ changes were observed.

Other findings:

none

Interpretation of results:

other: Not classified

Conclusions:

The acute oral LD50 value was considered to be >5000 mg//kg bw, when male and female Wistar rats were treated with the given test chemical via oral: gavage route.

Executive summary:

The acute oral toxicity study was conducted by using the given test chemical as per OECD Guideline 401 (Acute Oral Toxicity) in male and female Wistar rats at doses from 1000 and 5000 mg/kg bw.

The given test chemical was dissolved in polyethylene glycol (10-25%) and administered as 20 ml/kg via oral gavage route. Animals were observed for mortality five times during the day and daily thereafter. Body weights were recorded at the day of administration and days 7 and 14 after the administration. Necropsy of survivors was performed.

No mortality was observed at 5000 mg/kg bw in treated animals. Clinical signs were observed such as, Dyspnea, curved body position, diarrhoea and ruffeld fur at 1000 mg/kg and Sedation, dyspnea, exophthalmos, curved body position, diarrhoea and ruffeld fur at 5000 mg/kg. All animals had recovered within 7 observation days. Normal development in body weight was observed. No macroscopical organ changes were observed.

Therefore, the acute oral LD50 value was considered to be >5000 mg//kg bw, when male and female Wistar rats were treated with the given test chemical via oral: gavage route.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from study report.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Quality of whole database:

Waiver

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary source.

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

Acute dermal toxicity study was conducted by using the given test chemical.

GLP compliance:

not specified

Test type:

other: not specified

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rabbits were received from Pine Acres Rabbi try, West Brattleboro, Vermont
- Age at study initiation: These animals were 8-15 weeks old when obtained.
- Housing: The rabbits were individually housed in stainless steel cages,
- Diet (e.g. ad libitum): Fed Charles River Rabbit Formula (Agway) ad libitum and
- Water (e.g. ad libitum): provided untreated municipal water via water bottles
- Acclimation period: They were quarantined for two weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Animals were maintained at a temperature range of 67°F to 72°F
- Humidity (%): relative humidity range of 35% to 65%
- Air changes (per hr): Rooms are provided 12-16 air changes per hour
- Photoperiod (hrs dark / hrs light): Fluorescent lighting was controlled to provide a 12-hour
light cycle (1 AM to 7 PM).

Type of coverage:

occlusive

Vehicle:

other: 0.9% sodium chloride

Details on dermal exposure:

TEST SITE
- Area of exposure: The hair was carefully removed from the back and sides of each animal prior to testing. Oster animal clippers were used for this procedure.
- % coverage: 6 in. x 6 in
- Type of wrap if used: The trunk of the animal was wrapped in plastic wrap and then stockinette to prevent removal of the patches by the animal.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the wrappings and patches were removed.
- Time after start of exposure:24 hour

Duration of exposure:

24 hour

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

five male and five female

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed 3 times on the day of dosing and twice daily thereafter for signs of toxicity. Body weights were determined twice weekly.
- Necropsy of survivors performed: yes, necropsies were performed on animals dying intercurrently, as well as those surviving treatment and sacrificed on day 14.
- Other examinations performed: Histopathological examination of treated and untreated skin application sites was performed on animals dying during the test period and on 2 animals per sex necropsied at the end of the test period.

Statistics:

No data

Preliminary study:

No data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed at 2000 mg/kg bw during the 14 day observation period.

Clinical signs:

other: The only overt sign of toxicity observed during the study was mild diarrhea displayed by female rabbit 8850 in the afternoon of day 1. This mild diarrhea subsided completely by the morning of day 2.

Gross pathology:

There were no gross visible lesions detected upon necropsy of all 10 rabbits at the termination of the study.

Other findings:

No data

Interpretation of results:

other: Not classified

Conclusions:

The acute dermal LD50 value was considered to be >2000 mg/kg bw, when 10 male and female New Zealand White rabbits were treated with the given test chemical by dermal application.

Executive summary:

The acute dermal toxicity study was conducted by using the given test chemical in 10 male and female New Zealand White rabbits at the dose concentration of 2000 mg/kg bw. The hair was carefully removed from the back and sides of each animal prior to testing. Oster animal clippers were used for this procedure. The powdered test material for the single dose trials was applied uniformly to a 6 in. x 6 in. pad at a dose of 2000 mg/kg, moistened with 0.9% sodium chloride, and secured with surgical tape to the previously clipped and abraded test site. The trunk of the animal was wrapped in plastic wrap and then stockinette to prevent removal of the patches by the animal. After a 24 hour exposure period, the wrappings and patches were removed. All animals were observed 3 times on the day of dosing and twice daily thereafter for signs of toxicity. Body weights were determined twice weekly, and necropsies were performed on animals dying intercurrently, as well as those surviving treatment and sacrificed on day 14. Histopathological examination of treated and untreated skin application sites was performed on animals dying during the test period and on 2 animals per sex necropsied at the end of the test period. No mortality was observed at 2000 mg/kg bw during the 14 day observation period. The only overt sign of toxicity observed during the study was mild diarrhea displayed by female rabbit 8850 in the afternoon of day 1. This mild diarrhea subsided completely by the morning of day 2. The body weights of 3 male and 4 female rabbits either remained constant or increased during the study. Male rabbit 8828 lost approximately 100 grams between days 0 and 3, but regained weight by day 7. Female rabbit 8861 and male rabbit 8810 decreased in weight by 200 grams between days 10 and 14. There were no gross visible lesions detected upon necropsy of all 10 rabbits at the termination of the study.

Hence, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when 10 male and female New Zealand White rabbits were treated with the given test chemical by dermal application.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is from secondary source.

Additional information

Acute oral toxicity:

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for the given test chemical. The studies are summarized as below –

 

The reported study was mentioned in experimental study report and conducted to assess the toxicological profile of the given test chemical as per OECD Guideline 401 (Acute Oral Toxicity) in male and female Wistar rats at doses from 1000 and 5000 mg/kg bw. The given test chemical was dissolved in polyethylene glycol (10-25%) and administered as 20 ml/kg via oral gavage route. Animals were observed for mortality five times during the day and daily thereafter. Body weights were recorded at the day of administration and days 7 and 14 after the administration. Necropsy of survivors was performed. No mortality was observed at 5000 mg/kg bw in treated animals. Clinical signs were observed such as, Dyspnea, curved body position, diarrhoea and ruffled fur at 1000 mg/kg and Sedation, dyspnea, exophthalmos, curved body position, diarrhoea and ruffled fur at 5000 mg/kg. All animals had recovered within 7 observation days. Normal development in body weight was observed. No macroscopical organ changes were observed. Therefore, the acute oral LD50 value was considered to be >5000 mg//kg bw, when male and female Wistar rats were treated with the given test chemical via oral: gavage route.

 

The above study report is supported with another experimental study conducted by using the given test chemical. In an acute oral toxicity study, 10 female, 6 weeks old Wistar strain rats were given an oral dose of the given test chemical by gavage at a dose of 10000 mg/kg bw and 10 female, 6 weeks old Wistar strain rats were given an oral dose of test chemical by gavage at a dose of 15000 mg/kg bw. Both groups were observed for 14 days. During the observation period, no death, no clinical signs were observed in any animal. Thus, the lethal concentration (LD50) value for acute oral toxicity test was considered to be >15000 mg/kg bw, when female wistar W70 (SPF) rats were treated with the given test chemical orally via gavage.

 

This study is supported with the data available in authoritative database for the given test chemical. The acute oral toxicity study was performed by using the given test chemical in rats. Animals were observed for mortality. 50% mortality was observed at dose 3660 mg/kg bw. Hence, the acute oral LD50 value was considered to be 3660 mg/kg bw, when rats were treated with the given test chemical via oral route.

 

All these studies are further supported with the data available in secondary report for the given test chemical. The acute oral toxicity study was conducted in 10 male and female Fischer 344 rats at dose concentration of 5000 mg/kg bw. The given test chemical was dissolved in corn oil and administered via oral gavage route.  Animals were observed frequently on the day of dosing and twice daily thereafter for 14 days. Body weights were measured twice weekly.  Rats dying intercurrently as well as those surviving treatment and sacrificed on day 14 were necropsied. No mortality was observed at a dose of 5000 mg/kg to the rats throughout the 14 day observation period. By day 2 of the observation period, the fur on all 10 rats appeared green in colour. On day 4, the fur and tail of the male rats were green, and the fur and tail of the female rats were pale green. By day 14, the male rats were light green in colour. The females appeared light greenish/yellow by day 5, and yellow by day 9. By day 8, the males were light green in colour. All five male and four of the female rats gained or maintained weight throughout the course of the study. Female rat 11424 lost weight between days 7 and 11 but gained weight by day 14. Upon necropsy at terminus, there were no gross internal visible lesions detected in any of the rats. However, all five females were yellow in colour, and all five male rats were light green in colour. Therefore, the acute oral LD50 value was considered to be >5000 mg/kg bw, when 10 male and female Fischer 344 rats were treated with the given test chemical via oral gavage route.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

 

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to very low vapour pressure of the test chemical, which is reported to be 9.40E-11 Pa (7.05E-013 mmHg). Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal Toxicity:

In different studies, the given test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats and rabbits for the given test chemical. The studies are summarized as below -

 

The reported study was mentioned in secondary report to determine the acute dermal toxicity dose by using the given test chemical in 10 male and female New Zealand White rabbits at the dose concentration of 2000 mg/kg bw. The hair was carefully removed from the back and sides of each animal prior to testing. Oster animal clippers were used for this procedure. The powdered test material for the single dose trials was applied uniformly to a 6 in. x 6 in. pad at a dose of 2000 mg/kg, moistened with 0.9% sodium chloride, and secured with surgical tape to the previously clipped and abraded test site. The trunk of the animal was wrapped in plastic wrap and then stockinette to prevent removal of the patches by the animal. After a 24 hour exposure period, the wrappings and patches were removed. All animals were observed 3 times on the day of dosing and twice daily thereafter for signs of toxicity. Body weights were determined twice weekly, and necropsies were performed on animals dying intercurrently, as well as those surviving treatment and sacrificed on day 14. Histopathological examination of treated and untreated skin application sites was performed on animals dying during the test period and on 2 animals per sex necropsied at the end of the test period. No mortality was observed at 2000 mg/kg bw during the 14 day observation period. The only overt sign of toxicity observed during the study was mild diarrhoea displayed by female rabbit 8850 in the afternoon of day 1. This mild diarrhoea subsided completely by the morning of day 2. The body weights of 3 male and 4 female rabbits either remained constant or increased during the study. Male rabbit 8828 lost approximately 100 grams between days 0 and 3, but regained weight by day 7. Female rabbit 8861 and male rabbit 8810 decreased in weight by 200 grams between days 10 and 14. There were no gross visible lesions detected upon necropsy of all 10 rabbits at the termination of the study. Hence, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when 10 male and female New Zealand White rabbits were treated with the given test chemical by dermal application.

 

Another study mentioned in study report was carried out to determine the acute dermal toxicity dose by using the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Animals were observed for clinical signs, mortality, until sacrifice. Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behaviour pattern. Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14. Necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15). Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, LD50 was considered to be >2000mg/kg bw, when male and female Sprague Dawley rats was treated with test chemical by dermal application.

 

Both the above studies are further supported with the data mentioned in authoritative database by using the given test chemical in rabbits at the concentration of 2000 mg/kg bw. Animals were observed for mortality. No mortality was observed in treated rabbits at 2000 mg/kg bw. Therefore, LD50 value was considered to be >2000 mg/kg bw, when rabbits were treated with the given test chemical by dermal application to the skin.

 

Thus, based on the above summarised studies for test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies for the test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral toxicity and acute dermal toxicity. Thus, comparing these values with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral and acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.