3,3'-[(4-{(E)-[2-bromo-4-nitro-6-(trifluoromethyl)phenyl]diazenyl}phenyl)imino]dipropanenitrile

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 14-MAR-2000 to 26-MAY-2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guidance test with GLP compliance

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

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:

other: Directive 92/69/EEC, B.3. "Acute Toxicity-Dermal", July 31, 1992.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification: FAT41'029/A
Batch number: ARL 123/PP 1/99 UL
Purity: approx. 98 %
Color / Physical form: orange red to brown powder
Stability of test article: Stable under storage conditions
Expiry date: 04-NOV-2005
Stability of test article dilution: For at least 24 hours in polyethylene glycol at room temperature
Storage conditions: In the original container, at room temperature (range of 20 ± 3 °C), away from direct sunlight
Safety precautions: Routine hygienic procedures were used to ensure the health and safety of the personnel

Test animals

Species:

rat

Strain:

other: HanIbm: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST SYSTEM
Test system: Rat, HanIbm: WIST (SPF)
Source: RCC Ltd, Biotechnology & Animal Breeding Division, Wölferstrasse 4, CH-4414 Füllinsdorf / Switzerland
Number of animals per group: 5 males and 5 females
Age when treated: Males: 8-10 weeks; Females: 10-12 weeks
Body weight range when treated: Males: 212.4-219.3 g; Females: 179.8- 193.1 g
Acclimatization: One week under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

HUSBANDRY
Conditions
Standard Laboratory Conditions:
Air-conditioned with 10-15 air changes per hour and continuously monitored environment with a range for room temperature of 22-24 °C, and for relative humidity between 31-56 %. The animals were provided with a 12-hour artificial fluorescent light, 12-hour dark cycle. Music was played during the light period.
Accommodation: Groups of five in Makrolon type-4 cages with standard softwood bedding during the acclimatization. Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz) during treatment and observation.
Diet: Pelleted standard Kliba 3433, batch no.43/99 rat maintenance diet (Provimi Kliba AG, CH-4303 Kaiseraugst) available ad libitum.
Water: Community tap water from Füllinsdorf, available ad libitum.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Details on dermal exposure:

Twenty-four hours before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10 % of the total body surface. Only those animals without injury or irritation on the skin were used in the test.
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 and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
Application volume/kg body weight: 4.0 ml
Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed.

Duration of exposure:

Twenty-four hours

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

TEST ARTICLE PREPARATION
The test article was placed into a glass beaker on a tared Mettler PM 460 balance and the vehicle (PEG 300) was added. A weight by volume dilution was prepared using a magnetic stirrer as homogenizer. Homogeneity of the test article in the vehicle was maintained during treatment. The preparation was made shortly before each dosing.

OBSERVATIONS
Mortality / Viability: Four times during test day 1 and once daily during days 2-15.
Body weights: On test days 1 (pre-administration), 8 and 15.
Clinical signs: Each animal was examined for changes in appearance and behavior four times during day 1, and once daily during days 2-15. All abnormalities were recorded.

PATHOLOGY
NECROPSY
Necropsies were performed by experienced prosectors. At the end of the observation period all animals were sacrificed by intraperitoneal injection of NARCOREN (Rhône Mérieux GmbH, D-88471 Laupheim) at a dose of at least 2.0 ml/kg body weight (equivalent to at least 320 mg sodium pentobarbitone/kg body weight). The animals were examined macroscopically and all abnormalities recorded. Thereafter, they were discarded.

Statistics:

No statistical analysis was used.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: A light orange discoloration of the treated skin sites was observed in all animals after removal of the semi-occlusive dressing only.

Gross pathology:

No macroscopic findings were observed at necropsy.

Other findings:

no data

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The LD50 of FAT 41'029/A after single dermal administration to rats of both sexes, observed over a period of 14 days is, greater than 2000 mg/kg body weight.

Executive summary:

The acute dermal toxicity of  FAT41'029/A was performed according to OECD guideline No. 402 and Directive 92/69/EEC, B.3 under GLP compliance. A group of five male and five female HanIbm: WIST (SPF) rats was treated with FAT 41'029/A at 2000 mg/kg by dermal application. The test article was suspended in vehicle (polyethylene glycol, PEG 300) at a concentration of 0.5 g/ml and administered at a volume of 4 ml/kg. The animals were examined for local and/or clinical signs four times during day 1 and once daily during days 2-15. Mortality/viability were recorded together with clinical signs at the same time intervals. Body weights were recorded on day 1 prior to administration and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study. A light orange discoloration of the treated skin sites was observed in all animals after removal of the semi-occlusive dressing only. The body weight of the animals was within the range commonly recorded for animals of this strain and age. No macroscopic findings were observed at necropsy. Thus, the LD50 of FAT 41'029/A after single dermal administration to rats of both sexes, observed over a period of 14 days is, greater than 2000 mg/kg body weight.