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)

Acute Toxicity: dermal

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

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)

IUPAC Name:

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)

Test material form:

solid: particulate/powder

Details on test material:

- Description: Black powder
- Storage Conditions: Room temperature

Test animals

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.

Administration / exposure

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.

Results and discussion

Effect levels

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.

Applicant's summary and conclusion

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.