Trisodium bis[(3'-nitro-5'-sulfonato-(6-amino-2-[4-(2-hydroxy-1-naphtylazo)phenylsulfonylamino]pyrimidin-5-azo)benzene-2',4-diolato)]chromate(III)

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 26, 1994 - May 10, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

1992

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium bis[(3'-nitro-5'-sulfonato-(6-amino-2-[4-(2-hydroxy-1-naphtylazo)phenylsulfonylamino]pyrimidin-5-azo)benzene-2',4-diolato)]chromate(III)

EC Number:

418-220-4

EC Name:

Trisodium bis[(3'-nitro-5'-sulfonato-(6-amino-2-[4-(2-hydroxy-1-naphtylazo)phenylsulfonylamino]pyrimidin-5-azo)benzene-2',4-diolato)]chromate(III)

Molecular formula:

C52H32CrN18Na3O20S4

IUPAC Name:

chromium(3+) trisodium bis(6-amino-2-{4-[2-(2-hydroxynaphthalen-1-yl)diazen-1-yl]benzenesulfonamido}-5-[2-(3-nitro-2-oxido-5-sulfonatophenyl)diazen-1-yl]pyrimidin-4-olate)

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wñlferstrasse 4, CH-4414 Füllinsdorf/Switzerland
- Age at study initiation: males at 9 weeks; females at 11 weeks
- Weight at study initiation: males at 256.0 - 269.6; females at 189.0 - 209.7 g
- Diet: pelleted standard Kliba 343. Batch nos. 86/93 and 87/93 rat maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst) available ad libitum (except for over-night fasting period prior to application).
- Water: community tap water from Fülllnsdorf, available ad libitum.
- Acclimation period: one week under laboratory conditions» after health examination. Only animals without any visual signs of illness were used for the study.
- Identification: by unique cage number and corresponding color-coded spots on the tail.
- Randomization: randomly selected at time of delivery Into groups of five

ENVIRONMENTAL CONDITIONS
- Temperature: continuously monitored environment with a temperature of 22 ± 2 °C
- Humidity: 36 - 64 %
- Air changes: 10-15 ACH
- Photoperiod: 12 hours artificial fluorescent light (approx. 100 Lux) / 12 hours dark (light period between 6.00 a.m. to 6.00 p.m.), music during the light period
- Cage: during acclimatization in groups of five in Makrolon type-4 cages (size: 33 x 55 x 20 cm) with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz); during treatment and observation individually in Makrolon type-3 cages (size: 22 X 37.5 x 15 cm) with standard softwood bedding.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

bi-distilled

Details on dermal exposure:

TEST SITE
- Area of exposure: approximately 24 hours before treatment, the backs of the animals were clipped with an electric clipper. On test day 1, the test article was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe.
- % coverage: area of approximately 10 % of the total body surface.
- Type of wrap if used: semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
- Rationale: dermal administration was used because this is one possible route of human exposure during manufacture, handling and use of the test article.

REMOVAL OF TEST SUBSTANCE
- Washing: the skin was washed with lukewarm tap water and dried with disposable paper towels.
- Time after start of exposure: twenty-four hours after the application.

TEST MATERIAL
- Amount applied: 4.0 ml/kg
- For solids, paste formed: no
- Preparation: the test article was placed into a glass beaker on a tared Mettier PM 480 balance, and the vehicle, (bi-distilled water) was added. A weight by volume dilution was prepared using a homogenizer (Ultra-Turrax, Janke & Kunkel, D-79219 Staufen).
Homogeneity of the test article in the vehicle was maintained during treatment using a magnetic stirrer (Janke & Kunkel, D-79219 Staufen).
The preparation was made shortly before dosing.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw, single dose

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.
- Frequency of observations and weighing: on test day 1 (pre-administration), 8 and 15.
- Necropsy: necropsies were performed by experienced prosectors. At the end of the observation period all animals were anesthetized by intraperitoneal injection of NARCOREN at a dose of at least 2.0 ml/kg body weight and sacrificed by exsanguination. The animals were examined macroscopically.
- Other examinations performed: each animal was examined for changes in behaviour and appearance (with special emphasis on the application area, except for the time when the semi-occlusive dressing was in place) four times during day 1, and once dally during days 2-15.

Statistics:

The LOGIT-Model could not be used as no deaths occurred,

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Value not quantified since higher than tested concentrations

Mortality:

No deaths observed.

Clinical signs:

other: No clinical signs of systemic toxicity.

Gross pathology:

No organ abnormalities were observed at necropsy.

Other findings:

A red discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day 2 and persisted until test day 3 - 6.

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

LD50 (males and females) > 2000 mg/kg b.w.

Executive summary:

The test article was administered to a group of 5 male and 5 female rats by semi-occlusive dermal application with a single dose of 2000 mg test article/kg body weight.

- No deaths occurred during the study period.

- No clinical signs of systemic toxicity were observed during the observation period.

- A red discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day 2 and persisted until test day 3 - 6.

- There were no test article-related effects on the body weight of the animals during the observation period. The slight reduction in body weight gain during the first observation week was considered to be a consequence of the semi occlusive dressing.

- The macroscopic examination at study termination revealed no organ abnormalities.

In conclusion, the mean lethal dose after single oral administration to rats of both sexes, observed over a period of 14 days, could not be estimated, because no deaths occurred at the maximal dose of 2000 mg test article/kg body weight.