Black HM 2482

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 12th to February 15th, 1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1980

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted May 12, 1985

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

EWG Directive 84/449/ EWG, Amtsblatt der Europaeischen Gemein-schaften L 251, Jahrgang 27, 19. 9. 84. B.4 Akute Toxizitaet-Hautreizung / 106

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

400-920-6

EC Name:

-

Cas Number:

89857-06-7

Molecular formula:

C50 H53 N11 O14 S

IUPAC Name:

5'-[2-(7-{2-[4-(2-{1'-[3-(dimethylazaniumyl)propyl]-6'-hydroxy-4'-methyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium-5'-yl}diazen-1-yl)phenyl]diazen-1-yl}-8-hydroxy-6-sulfonatonaphthalen-2-yl)diazen-1-yl]-6'-hydroxy-3,4'-dimethyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium bis(2-hydroxypropanoate)

Test animals

Species:

rabbit

Strain:

New Zealand White

Remarks:

KFM

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG CH 4414 Fuellinsdorf /Switzerland
- Age at study initiation: 15 -16 weeks
- Weight at study initiation: 2.8 - 3.2 kg
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system
- Diet: pelleted standard KIiba 341, Batch 3/84 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, Switzerland), ad libitum
- Water: community tap water from Itingen, ad libitum
- Acclimation period: four days under test conditions after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Air changes: 10 -15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark, at least 8 hours music/light period

Test system

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

0.5 g test article

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48, and 72 hours after removing the gauze patches

Number of animals:

3 rabbits (2 males and 1 female)

Details on study design:

TEST SITE
- Area of exposure: 100 cm²
- Type of wrap if used: guaze patch covered with aluminium foil and then covered with an occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): four hours after the application, the dressing was removed and the skin was flushed with luke warm tap water
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
- Viability/Mortality/Toxicity: daily
- Body Weight: pre-test, day 1 and at termination of test on a Mettler PK 16 balance
- Symptoms: The skin reactions were assessed 1, 24, 48 and 72 hours intervals after the removal of the dressing, gauze patch and test article.
- Possible corrosive properties of the test article and the color to the treated skin area were recorded.

SCORING SYSTEM: according to OECD 404
Erythema and Eschar Formation
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beef redness) to slight eschar formation (injuries in depth): 4
Maximum possible: 4

Oedema Formation
No oedema: 0
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well defined by definite raising): 2
Moderate oedema (raised approximately 1 mm): 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure): 4
Maximum possible: 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Test item showed a primary irritation score of 0 when applied to intact rabbit skin.
No destructions or irreversible alterations of the treated skin were observed. No corrosion effect was evident on the skin.

Any other information on results incl. tables

COLORATION
In the area of application a severe black discoloration of the skin was observed which could be related to effects of the test article .

BODY WEIGHTS
The body weight gain of all rabbits was similar.

TOXIC SYMPTOMS/MORTALITY
No acute toxic symptoms were observed in the animals during the test period, and no mortality occurred.

NECROPSY
Due to the results obtainedr no macroscopic organ examination was indicated.

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

Not irritating

Executive summary:

The skin irritation potential of the substance, when single doses of the test item are placed on the skin of rabbits, was evaluated in an in vivo experimental study according to the OECD Guideline 404. On test day 1, 0.5 g of the test article was applied to the intact skin of the shaved area and the skin reaction was assessed at 1, 24, 48 and 72 hours intervals after the removing of the dressing, gauze patch and test article.
Under the conditions of this experiment, test item was found to cause a primary irritation score of 0 when applied to intact rabbit skin. In the area of application a severe black discoloration of the skin was observed which could be related to effects of the test article. No corrosion effect had occurred on the skin at each measuring interval.

The mean value of the scores for each type of lesion, calculated for each animal separately is the following:

Animal No.  Sex                Mean 24 -72 hours
                                      Erythema      Edema 
125            male                    0               0
126            male                    0               0
127            female                 0                0

In conclusion, it is reasonable to not classify the substance as skin irritant according to the CLP Regulation (EC 1272/2008).