A mixture of: disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(ethensulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex; disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(2-hydroxyethylsulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 November 1995 to 30 November 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

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:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

A mixture of: disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(ethensulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex; disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(2-hydroxyethylsulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex;

EC Number:

423-940-7

EC Name:

A mixture of: disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(ethensulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex; disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(2-hydroxyethylsulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex;

Cas Number:

85585-91-7

IUPAC Name:

4-[2-(7-{2-[4-(ethenesulfonyl)-2-hydroxy-5-methoxyphenyl]diazen-1-yl}-8-hydroxy-6-sulfonaphthalen-2-yl)diazen-1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid 4-[2-(8-hydroxy-7-{2-[2-hydroxy-4-(2-hydroxyethanesulfonyl)-5-methoxyphenyl]diazen-1-yl}-6-sulfonaphthalen-2-yl)diazen-1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid dicopper tetrasodium hydride

Test material form:

solid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Age at start of treatment: Approximately 8 weeks

Body weight at start of treatment: Within +/- of the sex mean

Number of animals: 5 males and 5 females

Identification: Earmark

Conditions
Air-conditioned room with approximately 15 air changes per hour amd the environment controlled with optimal conditions considered as being a temperature of 21°C and a relative humidity of 50%. Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity. Lighting was 12 hours artifical fluorescent light and 12 hours dark per day.

Accomodation
Individually housed in polycarbonate cages containing purified sawdust as bedding material. Certificates of analysis were examined and then retained in the NOTOX archives. Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.

Diet
Free access to standard pelleted laboratory animal diet. Certificates of analysis were examined and then retained in the NOTOX archives,

Water
Free access to tap water. Certificates of analysis (performed quarterly) were examined and then retained in the NOTOX archives,.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

Shaving
One day before exposure (Day 1) and area of approximately 5 x 7 cm on the back of the animal was clipped.

Application
The formulation was applied to an area of approximately 25 cm2 (5 x 5 cm) for males and 18 cm2 (3.5 x 5 cm) for females by application on a gauze pacth fixed succesively to aluminium foil and flexible bandage., with drops of petrolatum.

Frequency
Once, on Day 1

Application period
24 hours, thereafter dressings were removed and residual test substance was removed using a tissue moistened with tap water.

Duration of exposure:

24 hours

Doses:

2000 mg/kg

Volume: 10 ml/kg bw

No. of animals per sex per dose:

5 males, 5 females

Control animals:

no

Details on study design:

Observations

Mortality/ viability
Twice daily

Body weights
Days 1 (pre-administration), 8 and 15

Clinical signs
At periodic intervals on the day of treatment (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded.

Necropsy
At the end of the observation period, all animals were sacrificed by oxygen/ carbon dioxide asphyxiation and subjected to necropsy. Descriptions of all internal macropscopic abnormalities were recorded.

Statistics:

No statistical analyses were performed.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No animals died during the study

Clinical signs:

other: Red staining of the snout was observed among four animals between days 1 to 4 and in the neck region of one female on Day 12. During exposure, black staining of the neck region at the border of the bandages was apparent in the majority of animals and bla

Gross pathology:

No abnormalties were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of Pacified Reactive Black 31 in rats was established as exceeding 2000 mg/kg bw.

Executive summary:

The study was carried out in accordance with OECD Guideline No. 402, "Acute dermal toxicity" and EEC Directive 92/69/EEC, Part B.3, "Acute Toxicity - Dermal".

Pacified Reactive Black 31 was administered to five rats of each sex dy dermal application of 2000 mg/kg bw for 24 hours. Animals were subjected to daily observations and weekly determinatons of bodyweight. Macropscopic examination was performed at the the endof th experimental period - Day 15.

No animals died during the study

During exposure, black staining of the neck region at the border of the bandages was apparent in the majority of animals and black staining of the snout in one animal.

Black staining was seen in the treated skin-area of all animals during the observation period and persisted until Day 15.

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study amd were therefore considered not toxicologicaly significant.

No abnormalities were found in the animals at macroscopic post mrtem examination.

The dermal LD50 value of Pacified Reactive Black 31 in rats was established as exceeding 2000 mg/kg bw.