Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 May 2007 to 22 May 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

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
Test material form:
solid: particulate/powder
Details on test material:
Reactive Yellow 176 Ester
Reactive Yellow 176 Sulfato

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Species and strain: CRL:(WI)BR Wistar rats
Source: CHARLES RIVER (EUROPE) LABORATORIES INC. TOXI COOP Ltd. 1103 Budapest, Cserkesz u. 90.
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: The Wistar rat as a rodent is one of the standard species of acute toxicity studies
Number of animals: 5 animals/sex
Sex: Male and female; female rats were nulliparous and non-pregnant.
Age of animals: Young adult rats
Body weight range at the treatment: Male 241 - 248 g
Female 200 - 212 g
Animals arrived: 03 May 2007
Acclimatization time: 5 days
Animal health: Only healthy animals were used for the study. The veterinarian certified health status.
Number of animal room: 245/9
Housing: Individual caging
Cage type: Type II polypropylene/polycarbonate
Bedding: Laboratory bedding
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 8-12 air exchanges/hour by central air-condition system.
Food and Water Supply The 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 and tap water from municipal supply from 500 ml bottle, ad libitum.
The diet and drinking water are routinely analysed and are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Animal Identification: The individual identification was performed by numbers on the tail written with a permanent marker. The cages were marked by identity cards with information about study code, sex, dose group, cage number and individual animal number.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
moistened with 0.5 ml of distilled water.
Details on dermal exposure:
For the treatment the test item was applied in its original form moistened with 0.5 ml of distilled water.
The test item was not expected to be lethal at dermal 2000 mg/kg bw based on data provided by Sponsor. No test item related mortality occurred, thus full test was not conducted. A limit test was carried out involving a dose group of 2000 mg/kg bw with five animals/sex.

A single administration was performed by dermal route and was followed by a fourteen-day observation period.

The back of animals was shaven (approximately 10 % area of the total body surface) 24 hours prior to the treatment. The test item was applied in a single dose in original form uniformly over the shaved skin throughout a 24- hour exposure period. Sterile gauze pads were placed on the skin of rats. These gauzes were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was wrapped with semi occlusive plastic wrap for 24 hours.
At the end of the exposure period, residual test item was removed, using body temperature water.
Duration of exposure:
24 h
Doses:
Dose Group Number of animals
2000 mg/kg bw male 5
2000 mg/kg bw female 5
No. of animals per sex per dose:
Dose Group Number of animals
2000 mg/kg bw male 5
2000 mg/kg bw female 5

Control animals:
no
Details on study design:
Clinical Observations
Careful clinical observation was made at the following intervals: 1h, 5h after the treatment and once each day for 14 days thereafter.
Individual observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern as well. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement
The body weight of each animal was recorded on day 0 (shortly before the treatment), then on days 7 and 14 with precision of 1 g.

NECROPSY
At the end of the observation period gross pathology was conducted in every experimental animal. Animals were exsanguinated under pentobarbital anaesthesia. After examination of the external appearance, the cranial, thoracic, and abdominal cavities were opened and tissues, organs were observed. Gross pathological changes were recorded for each animal on the post mortem record sheets.
Statistics:
Frequency of the clinical signs and necropsy findings by sex were presented in tabular form.
The mean body weight and standard deviation were calculated.

Results and discussion

Preliminary study:
The test item was not expected to be lethal at dermal 2000 mg/kg bw based on data provided by Sponsor. No test item related mortality occurred, thus full test was not conducted. A full test needs to be considered, only if compound related mortality occurs (TGs OECD 402, Dir. 92/69/EC B.3 and OPPTS 870.1200.).
Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred after the 24-hour dermal exposure to Gelb Sulfato in CRL:(WI) BR rats.
Clinical signs:
No behavioural or dermal changes or general toxic symptoms were noted during the study.
The skin became yellow - red coloured after the patch removal, which disappeared between days 3 and 10.
Body weight:
The mean body weight and the body weight gain of animals were in the normal range during the two weeks observation period, similar to the expected values in untreated animals of the same age and strain.
Gross pathology:
Test item related macroscopic alterations were not found at the gross pathology. Pinprick-sized haemorrhages (2/5 male, 2/5 female) were observed in the lungs, which were caused by exsanguination procedures. Hydrometra due to the sexual cycle occurred in female animals (3/5).
Other findings:
None.

Any other information on results incl. tables

CLINICAL OBSERVATIONS

 

SUMMARY OF CLINICAL OBSERVATIONS

Group

Observations

Frequency

Male

Female

2000 mg/kg bw

Yellow – red coloured normal skin

5/5

5/5

Cessation of discolouration

Between days 3 and 10

Between days 3 and 9

Comment: Frequency = number of animals with symptom / number of animals observed

 

INDIVIDUAL CLINICAL OBSERVATIONS

Group

Animal Number

Observations

Day 0

Days

F

1h

5h

1

2

3

4

5

6

7

8

9

10

11-14

2000 mg/kg bw Male

5586

Normal

+

+

-

-

+

+

+

+

+

+

+

+

+

14/16

Yellow-red coloured skin

-

-

+

+

-

-

-

-

-

-

-

-

-

2/16

5587

Normal

+

+

-

-

+

+

+

+

+

+

+

+

+

14/16

Yellow-red coloured skin

-

-

+

+

-

-

-

-

-

-

-

-

-

2/16

5588

Normal

+

+

-

-

-

-

-

-

-

-

-

+

+

7/16

Yellow-red coloured skin

-

-

+

+

+

+

+

+

+

+

+

+

+

9/16

5589

Normal

+

+

-

-

-

-

+

+

+

+

+

+

+

12/16

Yellow-red coloured skin

-

-

+

+

+

+

-

-

-

-

-

-

-

4/16

5590

Normal

+

+

-

-

-

-

-

-

-

-

+

+

+

8/16

Yellow-red coloured skin

-

-

+

+

+

+

+

+

+

+

-

-

-

8/16

 

Group

Animal Number

Observations

Day 0

Days

F

1h

5h

1

2

3

4

5

6

7

8

9

10

11-14

2000 mg/kg bw Female

5631

Normal

+

+

-

-

-

-

-

+

+

+

+

+

+

11/16

Yellow-red coloured skin

-

-

+

+

+

+

+

-

-

-

-

-

-

5/16

5632

Normal

+

+

-

-

+

+

+

+

+

+

+

+

+

14/16

Yellow-red coloured skin

-

-

+

+

-

-

-

-

-

-

-

-

-

2/16

5633

Normal

+

+

-

-

+

+

+

+

+

+

+

+

+

14/16

Yellow-red coloured skin

-

-

+

+

-

-

-

-

-

-

-

-

-

2/16

5634

Normal

+

+

-

-

-

-

-

-

-

-

+

+

+

8/16

Yellow-red coloured skin

-

-

+

+

+

+

+

+

+

+

-

-

-

8/16

5635

Normal

+

+

-

-

+

+

+

+

+

+

+

+

+

14/16

Yellow-red coloured skin

-

-

+

+

-

-

-

-

-

-

-

-

-

2/16

Skin = Treated skin surface. It was intact in each case.

 

Comment:         F = Frequency of symptom = number of occurrence of symptom / total number of observations

                       + = Observation present, - = Observation not present; h = hours;

 

NECROPSY FINDINGS

 

TEST ITEM: GELB SULFATO

TEST SYSTEM: CRL: (WI) BR RAT

STUDY CODE: 07/425-002P

MODE OF ADMINISTRATION: DERMAL

SEX: MALE

DURATION OF STUDY: 14 DAYS

DOSE: 2000 MG/KG BW

NUMBER OF ANIMALS: 5/5

DATE OF NECROPSY: 22 MAY 2007

NECROPSY FINDINGS PER ORGANS

ANIMAL NUMBERS:

5586

5587

5588

5589

5590

NECROPSY FINDINGS

Σ

%

LUNGS

Pinprick-sized haemorrhages

-

-

-

+

+

2

40

NO ORGANS WITH MACROSCOPIC FINDINGS

+

+

+

-

-

3

60

 

TEST ITEM: GELB SULFATO

TEST SYSTEM: CRL: (WI) BR RAT

STUDY CODE: 07/425-002P

MODE OF ADMINISTRATION: DERMAL

SEX: FEMALE

DURATION OF STUDY: 14 DAYS

DOSE: 2000 MG/KG BW

NUMBER OF ANIMALS: 5/5

DATE OF NECROPSY: 22 MAY 2007

NECROPSY FINDINGS PER ORGANS

ANIMAL NUMBERS

5631

5632

5633

5634

5635

NECROPSY FINDINGS

Σ

%

LUNGS

Pinprick-size haemorrhages

-

+

+

-

-

2

40

UTERUS

Hydrometra

-

-

1

1

2

3

60

NO ORGANS WITH MACROSCOPIC FINDINGS

+

-

-

-

-

1

20

COMMENT:        NO ALTERATION = -

                               ALTERATION PRESENT = +

                               NO DATA = /

                               GRADE OF ALTERATION 1 = SLIGHT / SMALL / FEW

                                                              2 = MODERATE / M.SIZE / M. NUMBER

                                                              3 = MARKED / MANY / LARGE

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under our experimental conditions, the acute dermal LD50 value of the test item Gelb Sulfato was greater than 2000 mg/kg bw in male and female CRL: (WI) BR Wistar rat.
Gelb Sulfato does not meet the criteria for classification according to EU labelling regulations Commission Directive 2001/59/EC for classification and labelling of dangerous substances.
Executive summary:

The acute dermal toxicity of test item Gelb Sulfato was assessed in rats to provide information on health hazards likely to arise from 24-hour exposure by the dermal route.

 

A limit test according to OECD 402 and B.3. 92/69/EEC was performed with Gelb Sulfato in CRL: (WI) BR rats. The study was performed in accordance with the Principles of Good Laboratory Practice (GLP) and reported with a GLP certificate.

 

Five male and five female animals were treated with Gelb Sulfato for a single 24-hour dermal exposure in its original form at a dose level of 2000 mg/kg bw.

 

Clinical examinations were made on the day of treatment 1 h and 5 h after the application of the test item, and once each day for 14 days thereafter. The body weights of animals were recorded on day 0 and weekly thereafter. A gross necropsy was performed in all animals at the end of observation period.

 

Results

Mortality:None of the animals died.

 

Dose

2000 mg/kg bw

Male treated

5

Mortality

0/5

Female treated

5

Mortality

0/5

 

Clinical observations:Gelb Sulfato caused yellow – red discolouration of the treated skin for 2 - 9 days after the patch removal.

There were no behavioural changes or general toxic signs. Behaviour and general state of animals were considered to be normal.

 

Body weight and body weight gain:The body weight development was not influenced by the single dermal treatment with the test item Gelb Sulfato.

 

Necropsy:Test item related specific macroscopic alterations were not found.

 

Conclusions

Under the conditions of the present study, single 24-hour dermal administration of the test item, Gelb Sulfato did not cause mortality and toxic dermal alterations in male and female CRL:(WI)BR rats.

The acute dermal LD50 value of the test item Gelb Sulfato was greater than 2000 mg/kg bw in male and female CRL:(WI) BR rats.

Gelb Sulfato does not meet the criteria for classification according to EU labelling regulations for classification and labelling of dangerous substances.