Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 August 2011 to 07 September 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
On occasion during the study, the temperature (22 ± 3°C ) was recorded out of the target range. The actual range was at the temperature 20.7- 26.2 °C.
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Substance name: Reactive Yellow F01-0555

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
EXPERIMENTAL ANIMALS
Species and strain: CRL:(WI)BR Wistar rats
Source: CHARLES RIVER (EUROPE) LABORATORIES INC.
Hygienic level at arrival: SPF
Hygienic level during
the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies
Number of animals: 5 animals/sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at study start: Young adult rats
Body weight range
at dosing: Between 215 g and 280 g
Acclimatization time: 13 days

Husbandry
Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room-Box: 242/5
Housing: Individual caging
Cage type: Type II. polypropylene/polycarbonate
Bedding: Laboratory bedding:
Lignocel Hygienic Animal Bedding produced by J. Rettenmaier & Söhne GmbH+Co.KG (Holzmühle 1, 73494 Rosenberg, Germany);
A copy of the relevant Certificate of Analysis is maintained in CiToxLAB Hungary Ltd.'s archive.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20.7- 26.2 °C
Relative humidity: 44 - 70 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
The temperature and relative humidity was recorded twice daily during the study.

Food and Water Supply
Animals received ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is not considered to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
The supplier provided an analytical certificate for the batch used.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Identification
The individual identification was performed using numbers written on the tail with a marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File for each animal allocated to the treatment groups. The cages were identified by cards containing information about study code, sex, dose group, cage number and individual animal numbers.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment. The test item was applied as a single dose as supplied to the shaved skin and remained in contact with the skin for the 24- hour exposure period. For that purpose, the appropriate amount of the test item was moistened with body temperature water and distributed as uniformly as possible. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.
At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
Justification of the dose: The test item was not expected to be lethal at 2000 mg/kg bw. A limit test was therefore performed.
No. of animals per sex per dose:
5 male + 5 female
Control animals:
no
Details on study design:
Clinical Observations
Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Measurement of Body Weight
The body weights were recorded on Day 0 (before test item administration) and on Days 3, 7 and 14.

NECROPSY
All animals were anaesthetised with Euthasol®40% (details in 3.1.3.) and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.
Statistics:
None

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
No mortality occurred.
Clinical signs:
No clinical signs were observed after the treatment with the test item or during the 14-day observation period.
Body weight:
One female showed slight bodyweight loss during the first week of the observation period.
Gross pathology:
There was no evidence of test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Pelvic dilation of the right kidney was seen as an incidental finding in one male rat (7931).
Other findings:
No local dermal signs were observed during the entire study period. However, orange staining was observed on the skin in all animals after dosing from Day 1 to Day 8.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item Reactive Yellow F01-0555 was found to be higher than 2000 mg/kg bw in male and female CRL:(WI)BR Wistar rats.
Executive summary:

An acute dermal toxicity study was performed with test item Reactive Yellow F01-0555 in CRL:(WI)BR Wistar rats, in compliance with OECD Guideline No.: 402. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied, moistened with distilled water, as a single dermal 24-hour exposure followed by a 14-day observation period. Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 3, 7 and 14. Rats were euthanized and a gross macroscopic examination performed at the end of the 2-week observation period (Day 14).

The results of the study were summarized as follows:

Mortality: No mortality occurred.

Systemic clinical signs: No clinical signs were observed after the treatment with the test item or during the 14-day observation period. Local dermal signs: No local dermal signs were observed during the entire study period. However, orange staining was observed on the skin in all animals after dosing from Day 1 to Day 8.

Body weight: One female showed slight bodyweight loss during the first week of the observation period.

Necropsy: There was no evidence of test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Pelvic dilation of the right kidney was seen as an incidental finding in one male rat (7931).

The acute dermal median lethal dose (LD50) of the test item Reactive Yellow F01-0555 was found to be higher than 2000 mg/kg bw in male and female CRL:(WI)BR Wistar rats.