Reaction mass of hydrogen [1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphtholato(2-)][1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) , compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1) and hydrogen bis[1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) , compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1) and hydrogen bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) , compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1)

Endpoint summary

Description of key information

Acute Oral Toxicty: Under the conditions of the study, the acute lethal oral dose was greater than 5 000 mg/kg bodyweight in male and female rats.

Acute Dermal Toxicity: Under the conditions of the study, the acute LD50 of the test material in the Sprague-Dawley rat was greater than 2 000 mg/kg bodyweight.

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:

1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test material was prepared at a concentration of 50 % w/v in corn oil and administered at a volume of 10.0 mL/kg. The test material was prepared on the day of dosing.

Species:

rat

Strain:

other: Crl:CD (SD) BR VAF plus

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Approximately four to six weeks of age.
- Weight at study initiation: Weight range of 100 to 123 g prior to dosing.
- Fasting period before study: Yes. Access to food only was prevented overnight prior to and approximately 4 hours after dosing.
- Housing: Housed in groups of up to five rats of the same sex in metal cages with wire mesh floors.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: All the rats were acclimated to the experimental environment for a period of 6 days prior to the start of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 °C to 25 °C
- Humidity (%): 60 % R.H
- Air changes (per hr): The rate of air exchange was maintained at approximately 15 air changes/hour.
- Photoperiod (hrs dark / hrs light): Lighting was controlled by means of a time switch to provide 12 hours artificial light in each 24-hour period.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test material was prepared at a concentration of 50 % w/v in corn oil.

MAXIMUM DOSE VOLUME APPLIED:
The test material was administered at a volume of 10.0 mLkg.

Doses:

5.0 g/kg bodyweight. The appropriate dose volume of the test substance was administered to each rat using a syringe and plastic catheter (10 choke).

No. of animals per sex per dose:

Five males and five females.

Control animals:

no

Details on study design:

- Duration of observation period following administration: All animals were observed for 14 days after dosing. The day of dosing was designated Day 1.
- Frequency of observations and weighing: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1 (a period of five hours). On subsequent days the animals were observed once in the morning and again at the end of the experimental day. Clinical signs were recorded at each observation. The nature, severity, approximate time of onset and duration of each toxic sign.
Individual bodyweights of rats were recorded on Days 1 (day of dosing), 8 and 15.
- Necropsy of survivors performed: Yes. All animals were killed on Day 15 by cervical dislocation and were subjected to a macroscopic post mortem examination which consisted of opening the abdominal and thoracic cavities. The macrospic appearance of all examined tissues were recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths following a single oral dose of test material at 5.0 g/kg bodyweight.

Clinical signs:

other: Pilo-erection was observed in all rats within five minutes of dosing and throughout the remainder of Day 1. Dark blue eyes and extremities were observed in all rats from Day 3 to Day 6 or 7. Abnormal body carriage (hunched posture) was also observed in al

Gross pathology:

Terminal autopsy findings were normal.

Interpretation of results:

other: Not classified in accordance with EU criteria.

Conclusions:

Under the conditions of the study, the acute lethal oral dose was greater than 5 000 mg/kg bodyweight in male and female rats.

Executive summary:

The acute oral toxicity of the test material was assessed in the rat in accordance with the standardised guideline EU Method B.1 under GLP conditions.

A group of ten rats (five males and five females) was treated with test material at 5 000 mg/kg bodyweight. The appropriate dose volume of the test substance was administered to each rat using a syringe and plastic catheter (10 choke). All animals were observed for clinical signs and mortality for a period of 14 days after dosing. Individual bodyweight gains were also monitored. All animals were euthanised on Day 15 by cervical dislocation and were subjected to a macroscopic post mortem examination.

There were no deaths following a single oral dose of test material at 5 000 mg/kg bodyweight. Pilo-erection was observed in all rats within five minutes of dosing and throughout the remainder of Day 1. Dark blue eyes and extremities were observed in all rats from Day 3 to Day 6 or 7. Abnormal body carriage (hunched posture) was also observed in all females during this period. Recovery, as judged by external appearance and behaviour, was complete by Day 9. Slightly low bodyweight gains during the first week of the study were recorded for one male and three female rats on Day 8. All other rats achieved anticipated bodyweight gains throughout the study. Terminal autopsy findings were normal.

Under the conditions of the study, the acute lethal oral dose to rats of the test material was found to be greater than 5 000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

The key study was conducted under GLP conditions in accordance with the standardised guideline EU method B1. It was assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch (1997).

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:

06 January 1993 to 20 January 1993.

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

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
- Age at study initiation: Approximately ten to fourteen weeks old.
- Weight at study initiation: Male rats weighed 216 - 252 g and female rats weighed 209 - 228 g.
- Housing: The animals were housed in suspended polypropylene cages furnished with softwood sawdust. The animals were housed individually during the 24-hour exposure period and in groups of five, by sex, for the remainder of the study.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: At least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 21 °C
- Humidity (%): 52 - 67 %
- Air changes (per hr): Approximately 15 changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Remarks:

The test material was used as supplied.

Details on dermal exposure:

TEST SITE
- Area of exposure: 5 cm x 4 cm of dermal surface area
- % coverage: approximating to 10 % of the total body surface area.
- Type of wrap if used: A piece of surgical gauze measuring 7 cm x 4 cm was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage (HYPERTIE). The bandage was further secured with a piece of BLENDERM wrapped around each end.

REMOVAL OF TEST SUBSTANCE
- Washing: The bandage was carefully removed and the treated skin and surrounding hair were wiped with cotton wool moistened with arachis oil to remove any residual test material.
- Time after start of exposure: After the 24-hour contact period.

TEST MATERIAL
- For solids, paste formed: No. The test material was applied uniformly to an area of shorn skin which had previously been moistened with arachis oil B.P.

Duration of exposure:

24 hours

Doses:

2 000 mg/kg

No. of animals per sex per dose:

Five males and five females.

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: The animals were observed for deaths or overt signs of toxicity at 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days.
- Frequency of weighing: Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: At the end of the study the animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
- Other examinations performed: Adverse dermal reactions, if present, were recorded. Data evaluations included the relationship, if any, between the animals' exposure to the test material and the increased incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.
- Other: On the day before treatment, the back and flanks of each animal were clipped free of hair using veterinary clippers.

Statistics:

Using the mortality data obtained, an estimate of the acute dermal median lethal dose (LD50) of the test material was made.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths throughout the duration of the test.

Clinical signs:

other: No signs of systemic toxicity were noted during the study. Black staining of the fur was commonly noted during the study.

Gross pathology:

No abnormalities were noted at necropsy.

Other findings:

Dermal reactions: Desquamation was noted at the treatment sites of all animals four and five days after dosing and persisted at the treatment sites of two animals up to seven days after dosing. No other signs of skin irritation were noted.

Interpretation of results:

other: Not classified in accordance with EU criteria.

Conclusions:

Under the conditions of the study, the acute LD50 of the test material in the Sprague-Dawley rat was greater than 2 000 mg/kg bodyweight.

Executive summary:

A study was conducted to investigate the potential of the test material to cause acute dermal toxicity in rats in accordance to the standardised guidelines OECD 402 and EU method B3 under GLP conditions.

A group of ten animals (five males and five females) were given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2 000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment and were then euthanised for gross pathological examination.

There were no deaths throughout the study and no signs of systemic toxicity were noted during the study. Desquamation was noted at the treatment sites of all animals. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

Under the conditions of the study, the acute dermal LD50 of the test material in the Sprague-Dawley rat was found to be greater than 2 000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The key study was conducted under GLP conditions in accordance with the standardised guidelines OECD 402 and EU method B3. It was assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch (1997).

Additional information

Acute Oral Toxicity

The acute oral toxicity of the test material was assessed in the rat in accordance with the standardised guideline EU Method B.1 under GLP conditions. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

A group of ten rats (five males and five females) was treated with test material at 5 000 mg/kg bodyweight. The appropriate dose volume of the test substance was administered to each rat using a syringe and plastic catheter (10 choke). All animals were observed for clinical signs and mortality for a period of 14 days after dosing. Individual bodyweight gains were also monitored. All animals were euthanised on Day 15 by cervical dislocation and were subjected to a macroscopic post mortem examination.

There were no deaths following a single oral dose of test material at 5 000 mg/kg bodyweight. Pilo-erection was observed in all rats within five minutes of dosing and throughout the remainder of Day 1. Dark blue eyes and extremities were observed in all rats from Day 3 to Day 6 or 7. Abnormal body carriage (hunched posture) was also observed in all females during this period. Recovery, as judged by external appearance and behaviour, was complete by Day 9. Slightly low bodyweight gains during the first week of the study were recorded for one male and three female rats on Day 8. All other rats achieved anticipated bodyweight gains throughout the study. Terminal autopsy findings were normal.

Under the conditions of the study, the acute lethal oral dose to rats of the test material was found to be greater than 5 000 mg/kg bodyweight.

Acute Dermal Toxicty

A study was conducted to investigate the potential of the test material to cause acute dermal toxicity in rats in accordance to the standardised guidelines OECD 402 and EU method B3 under GLP conditions. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

A group of ten animals (five males and five females) were given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2 000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment and were then euthanised for gross pathological examination.

There were no deaths throughout the study and no signs of systemic toxicity were noted during the study. Desquamation was noted at the treatment sites of all animals. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

Under the conditions of the study, the acute dermal LD50 of the test material in the Sprague-Dawley rat was found to be greater than 2 000 mg/kg bodyweight.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to acute toxicity via the oral or dermal route.