Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 December 2010 to 22 December 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to EU, OECD & US EPA 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:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Product name: Reactive Red F08-0146

Test animals

Species:
rat
Strain:
other: Wistar RjHan:WI
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 214 g and 248 g
Acclimatization time: 6 days
Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room-Box: 244/9
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 Rosenberger, Germany).
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 21.4- 24.6°C
Relative humidity: 31 - 58 %
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).

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:
water
Details on dermal exposure:
The test item was applied as supplied moistened with distilled water.

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.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Males 5, Females 5
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 7 and 14.

Necropsy
All animals were anaesthetised with Euthasol 40% 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

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 of REACTIVE RED F08-0146 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 F08-0146 treated animals did not show any test item-related effect.
Gross pathology:
A single 24 hour dermal application of REACTIVE RED F08-0146 at a dose level of 2000 mg/kg bw was not accompanied with any macroscopic test item-related findings.
Other findings:
Red staining was recorded on the skin in all animals after dosing. The discoloration of the skin lasted up to 5 days in all animals. No other local dermal signs were observed.

Any other information on results incl. tables

INDIVIDUAL CLINICAL OBSERVATIONS

STUDY CODE: 10/285-002P          TEST SYSTEM: Rj Han: WI Rat      TEST ITEM: Reactive Red F08-0146

 

DOSE LEVEL: 2000 mg/kg                                                                                                                           SEX: MALE

Cage No.

Animal No.

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

1

3349

Symptom Free

+

+

-

-

-

-

-

+

+

+

+

+

+

+

+

+

11/16

Red skin

-

-

+

+

+

+

+

-

-

-

-

-

-

-

-

-

5/16

2

3350

Symptom Free

+

+

-

-

-

-

+

+

+

+

+

+

+

+

+

+

12/16

Red skin

-

-

+

+

+

+

-

-

-

-

-

-

-

-

-

-

4/16

3

3351

Symptom Free

+

+

-

-

-

-

+

+

+

+

+

+

+

+

+

+

12/16

Red skin

-

-

+

+

+

+

-

-

-

-

-

-

-

-

-

-

4/16

4

3352

Symptom Free

+

+

-

-

-

-

-

+

+

+

+

+

+

+

+

+

11/16

Red skin

-

-

+

+

+

+

+

-

-

-

-

-

-

-

-

-

5/16

5

3353

Symptom Free

+

+

-

-

-

-

+

+

+

+

+

+

+

+

+

+

12/16

Red skin

-

-

+

+

+

+

-

-

-

-

-

-

-

-

-

-

4/16

DOSE LEVEL:

2000 mg /kg                                                                                                                                    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

5h

6

3359

Symptom Free

+

+

-

-

-

-

-

+

+

+

+

+

+

+

+

+

11/16

Red skin

-

-

+

+

+

+

+

-

-

-

-

-

-

-

-

-

5/16

7

3360

Symptom Free

+

+

-

-

-

-

-

+

+

+

+

+

+

+

+

+

11/16

Red skin

-

-

+

+

+

+

+

-

-

-

-

-

-

-

-

-

5/16

8

3361

Symptom Free

+

+

-

-

-

-

-

+

+

+

+

+

+

+

+

+

11/16

Red skin

-

-

+

+

+

+

+

-

-

-

-

-

-

-

-

-

5/16

9

3362

Symptom Free

+

+

-

-

-

-

+

+

+

+

+

+

+

+

+

+

12/16

Red skin

-

-

+

+

+

+

-

-

-

-

-

-

-

-

-

-

4/16

10

3363

Symptom Free

+

+

-

-

-

-

+

+

+

+

+

+

+

+

+

+

12/16

Red skin

-

-

+

+

+

+

-

-

-

-

-

-

-

-

-

-

4/16

Remarks:            +: present

                           h=hour (s)          Treatment day= Day 0

                           Frequency of observation = number of occurrence of observation / total number of observations

 

BODY WEIGHT DATA

INDIVIDUAL BODY WEIGHT AND BODY WEIGHT GAIN

STUDY CODE: 10/285-002P          TEST SYSTEM: Rj Han: WI Rat      TEST ITEM: Reactive Red F08-0146

 

DOSE LEVEL: 2000 mg/kg                                                                                        SEX: MALE

Cage No.

Animal No.

Body weight (g) Days

Body Weight Gain (g)

0

7

14

0-7

7-14

0-14

1

3349

235

281

328

46

47

93

2

3350

248

300

361

52

61

113

3

3351

240

287

344

47

57

104

4

3352

241

288

336

47

48

95

5

3353

242

303

366

61

63

124

Mean:

241.2

291.8

347.0

50.6

55.2

105.8

Standard deviation:

4.7

9.3

16.2

6.3

7.4

12.9

DOSE LEVEL: 2000 mg/kg                                                                               SEX: FEMALE

Cage No.

Animal No.

Body weight (g) Days

Body Weight Gain (g)

0

7

14

0-7

7-14

0-14

6

3359

214

223

228

9

5

14

7

3360

230

241

260

11

19

30

8

3361

220

235

244

15

99

24

9

3362

218

219

238

1

19

20

10

3363

216

222

236

6

14

20

Mean:

219.6

228.0

241.2

8.4

13.2

21.6

Standard deviation:

6.2

9.5

12.0

5.3

6.2

5.9

Remarks:            Treatment day = Day 0

 

MACROSCOPIC FINDINGS

INDIVIDUAL INTERNAL AND EXTERNAL MACROSCOPIC OBSERVATIONS

STUDY CODE: 10/285-002P          TEST SYSTEM: Rj Han: WI Rat      TEST ITEM: Reactive Red F08-0146

 

DOSE LEVEL: 2000 mg/kg                                                                                                          SEX: MALE

Cage No.

Animal No.

External Observations

Internal Observations

Organ / Tissue

1

3349

No external observations

No internal observations

Not applicable

2

3350

No external observations

Dilation, pelvis, right

Kidneys

3

3351

No external observations

Dilation, pelvis, right

Kidneys

4

3352

No external observations

No internal observations

Not applicable

5

3353

No external observations

No internal observations

Not applicable

DOSE LEVEL: 2000 mg/kg                                                                                              SEX: FEMALE

Cage No.

Animal No.

External Observations

Internal Observations

Organ / Tissue

6

3359

No external observations

No internal observations

Not applicable

7

3360

No external observations

No internal observations

Not applicable

8

3361

No external observations

No internal observations

Not applicable

9

3362

No external observations

No internal observations

Not applicable

10

3363

No external observations

No internal observations

Not applicable

 

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 F08-0146 was found to be higher than 2000 mg/kg body weight in male and female RjHan:(WI) Wistar rats.
Executive summary:

This study has been performed in accordance with the study plan, OECD Guidelines for Testing of Chemicals (No.: 402, 24th Feb. 1987), Commission Regulation (EC) No 440/2008, B.3 (L142, 30 May 2008), OPPTS 870.1200 (EPA 712-C-98-192, August 1998) and the Principles of Good Laboratory Practice (GLP) and reported with a valid GLP certificate.

 

An acute dermal toxicity study was performed with test item REACTIVE RED F08-RjHan:(WI) Wistar rats.

 

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 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 5.

Body weight: The body weight and body weight gain of REACTIVE RED F08-0146 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 at necropsy. Pelvic dilation of the right kidney was seen incidentally.

 

Conclusions: The acute dermal median lethal dose (LD50) of the test item REACTIVE RED F08-0146 was found to be higher than 2000 mg/kg bw in male and female RjHan:(WI) Wistar rats.