Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-10-27 till 2009-11-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study under GLP without deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): FAT 40'849/A TE
- Substance type: colouring dye
- Physical state: blue powder
- Analytical purity: 76.9% of all coloured components
- Lot/batch No.: TZ 5463 / BOP 03-09
- Expiration date of the lot/batch: 2014-06-30
- Storage condition of test material: At room temperature at about 20 °C

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS RccHan: WIST(SPF)
- Source: Harlan Laboratories B.V. Kreuzelweg 53 5961 NM Horst / The Netherlands Postbus 6174 5960 AD Horst / The Netherlands
- Age at study initiation: Males: 9 weeks, Females: 11 weeks
- Weight at study initiation: Males 242.0 - 260.6 g ; Females: 190.6 - 207.9g
- Fasting period before study: no data
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 30/09 (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) ad libitum.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): relative humidity between 30-70% (values above 70% during cleaning process possible)
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
deionised water which was processed and treated by the PURELAB Option-R unit, which links four purification technologies: reverse osmosis, adsorption, ion-exchange and photo oxidation.
Details on dermal exposure:
TEST SITE
- Area of exposure: back of the animals
- % coverage: approx. 10% of the body surface
- Type of wrap if used: skin with a syringe and covered with a surgical gauze pad (ca. 5 x 5 cm) held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and an elastic adhesive restrainer bandage wrapped around the abdomen.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, flushed with lukewarm water and drapped off with disposable paper towels.
- Time after start of exposure: 24h

TEST MATERIAL AND VEHICLE
- Amount(s) applied (volume or weight with unit): 4mL/kg
- Concentration (if solution): dose: 2000mg/kg BW
- Constant volume or concentration used: yes
- test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle was added (weight:volume).

Duration of exposure:
24h
Doses:
2000mg/kg BW
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:

Viability / Mortality: Daily during the acclimatization period, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.

Clinical Signs: Daily during the acclimatization period, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.

Local Dermal Signs: Once daily during days 2 (following dressing removal) through day 15 using the numerical scoring system described in Appendix I.

Body Weights: On test days 1 (prior to administration), 8 and 15.

- Necropsy of survivors performed: yes
- Other examinations performed: macroscopic examinations were performed. An external examination and opening of the abdominal and thoracic cavities for examinations of major organs were performed. The appearance of any macroscopic abnormalities was recorded. No organs or tissues were retained.
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.
Remarks on result:
other: No statistical analysis was used.
Mortality:
No intercurrent deaths occurred during the course of the study.
Clinical signs:
No clinical signs were recorded in any of the animals throughout the complete observation
period.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necropsy.
Other findings:
Local Dermal Signs
In all animals, the moderate blue staining prevented the assessment of a possible erythema on test day 2 and 3. When assessable, only slight erythema was noted in one animal between test day 7 and 10. Additionally scaling was observed in the same animal between test day 8 and 10.
No local dermal sighs were observed in the other animals.

Test item Coloration
Slight to marked blue staining produced by the test item was observed immediately after removal of the dressing in all animals and persisted on the treated skin area up to test day 8 in six animals, up to test day 12 in two animals and up to test day 15, the end of the observation period, in two animals.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The median lethal dose of FAT 40849/A TE after single dermal administration to rats of both sexes, observed over a period of 14 days, is:
LD50 (rat): greater than 2000 mg/kg body weight
Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, FAT 40849/A TE does not have to be classified in respect to the acute dermal toxicity study in rats.
Executive summary:

Five male and five female RccHan:WIST (SPF) rats were treated with FAT 40849/A TE at 2000 mg/kg by dermal application. The test item was formulated in purified water at a concentration of 0.5 g/mL and administered at a volume dosage of 4 mL/kg. The application period was 24 hours.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Local signs were noted once daily from test day 2 to 15. Mortality/viability was recorded within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

No intercurrent deaths occurred during the course of the study.

No clinical signs were recorded in any of the animals throughout the complete observation period.

Slight to marked blue staining produced by the test item was observed immediately after removal of the dressing in all animals and persisted on the treated skin area up to test day 8 in six animals, up to test day 12 in two animals and up to test day 15, the end of the observation period, in two animals.

In all animals, the moderate blue staining prevented the assessment of a possible erythema on test day 2 and 3. When assessable, only slight erythema was noted in one animal between test day 7 and 10. Additionally scaling was observed in the same animal between test day 8 and 10. No local dermal sighs were observed in the other animals.

The body weight of the animals was within the range commonly recorded for this strain and age.

No macroscopic findings were observed at necropsy.

The median lethal dose of FAT 40849/A TE after single dermal administration to rats of both sexes, observed over a period of 14 days, is:

LD50 (rat): greater than 2000 mg/kg body weight

Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, FAT 40849/A TE does not have to be classified in respect to the acute dermal toxicity study in rats.