Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 February 2011 to 9 March 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to EU, US & OECD test guidance in compliance with GLP and reported with a valid GLP certificate.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
the humidity (30 – 70%) was recorded out of the target range. The actual range was at the humidity 24-55 %.
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
the humidity (30 – 70%) was recorded out of the target range. The actual range was at the humidity 24-55 %.
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
the humidity (30 – 70%) was recorded out of the target range. The actual range was at the humidity 24-55 %.
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: Reactive Red F03-0318

Test animals

Species:
rat
Strain:
other: RjHan:(WI) Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Species and strain: RjHan:(WI) Wistar rats
Source: Laboratoire Elevage Janvier, B.P. 4105, Route des Chênes Secs, 53940 Le Genest-St-Isle CEDEX FRANCE
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 205 g and 240 g
Acclimatization time: 6 days

Husbandry
Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room-Box: 245/5
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);
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 19.4- 23.8 °C
Relative humidity: 24 - 55 %
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.

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 LAB Research 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 LAB Research 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:
TEST SITE
- Area of exposure: approximately 10 % area of the total body surface
- % coverage: approximately 10 % area of the total body surface)
- Type of wrap if used: semi occlusive plastic wrap

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
- Time after start of exposure: 14 days

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): Not applicable
- Constant volume or concentration used: no. The appropriate amount of the test item was moistened with distilled water and distributed as uniformly as possible.
Duration of exposure:
14 days
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
DOSAGES

Justification of the doses:
The test item was not expected to be lethal at 2000 mg/kg bw. A limit test was therefore performed.

PROCEDURE

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 distilled 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.

OBSERVATIONS

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 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.

EVALUATION

Body weight and body weight gain are summarized in tabular form. Clinical signs and necropsy findings are described and summarized in tabular form.

Statistics:
None

Results and discussion

Preliminary study:
Not applicable
Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred after a 24-hour dermal exposure to Reactive Red F03-0318 administered at 2000 mg/kg bw to RjHan:(WI) Wistar rats followed by a 14-day observation period.
Clinical signs:
No clinical signs were observed after the treatment with the test item or during the 14 day observation period.
Body weight:
The body weight and body weight gain of Reactive Red F03-0318 treated animals did not show any test item-related effect.
Gross pathology:
At necropsy, no macroscopic test item-related findings were observed.
Other findings:
Red staining was recorded on the skin in all animals after dosing. The discoloration of the skin lasted up to Day 12 in all animals. No other local dermal signs were observed.

Any other information on results incl. tables

CLINICAL OBSERVATIONS

Dose level: 2000 mg/kg bw

Sex: Female

Cage No.

Animal No.

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5 h

1

4903

Symptom Free

+

+

 

 

 

 

 

 

 

 

 

 

 

+

+

+

5/16

Dark discoloration, red

-

-

+

+

+

+

+

+

+

+

+

+

+

-

-

-

1 1/16

2

4904

Symptom Free

+

+

 

 

 

 

 

 

 

 

 

 

 

+

+

+

5/16

Dark discoloration, red

-

-

+

+

+

+

+

+

+

+

+

+

+

-

-

-

1 1/16

3

4905

Symptom Free

+

+

 

 

 

 

 

 

 

 

 

 

 

 

+

+

4/16

Dark discoloration, red

-

-

+

+

+

+

+

+

+

+

+

+

+

+

-

-

12/16

4

4906

Symptom Free

+

+

 

 

 

 

 

 

 

 

 

 

 

+

+

+

5/16

Dark discoloration, red

-

-

+

+

+

+

+

+

+

+

+

+

+

-

-

-

1 1/16

5

4907

Symptom Free

+

+

 

 

 

 

 

 

 

 

 

 

 

 

+

+

4/16

Dark discoloration, red

-

-

+

+

+

+

+

+

+

+

+

+

+

+

-

-

12/16

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal median lethal dose (LD50) of the test item Reactive Red F03 0318 was found to be higher than 2000 mg/kg body weight in male and female RjHan:(WI) Wistar rats.
Executive summary:

An acute dermal toxicity study was performed with test item Reactive Red F03-inRjHan:(WI) 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, red staining was observed on the skin in all animals after dosing from Day 1 to Day 12.

Body weight

The body weight and body weight gain of Reactive Red F03-0318 treated animals did not show any test item-related effect.

 

Necropsy

There was no evidence of test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Uterus in estrus was observed in two females.


Conclusions

 

The acute dermal median lethal dose (LD50) of the test item Reactive Red F03‑0318 was found to be higher than 2000 mg/kg bw in male and female RjHan:(WI) Wistar rats.