[No public or meaningful name is available]

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July from 12 to 15, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 24 April 2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Direct Blue 267

IUPAC Name:

Direct Blue 267

Specific details on test material used for the study:

For the purpose of the study the test material was ground to a powder prior to application.
The pH of the test material (10 % w/w aqueous preparation) was determined prior to commencement of the study and found to be 6.6.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd, Moston, Sandbach, Cheshire, UK.
- Age at study initiation: twelve to twenty weeks old.
- Weight at study initiation: weight range of 2.0 to 3.5 kg.
- Housing: animals were individually housed in suspended cages.
- Diet: certified rabbit diet (Code 5322) supplied by BCM IPS Limited, ad libitum.
- Water: drinking water, ad libitum.
- Acclimation period: at least five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 17 to 23 °C
- Humidity: 30 to 70 %
- Air changes: at least fifteen changes per hour.
- Photoperiod: lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 g of test material, moistened with 0.5 ml of distilled water

Duration of treatment / exposure:

4-hours

Observation period:

72 hours

Number of animals:

3 rabbits

Details on study design:

TEST SITE
- Area of exposure: on the day before the test each of a group of three rabbits was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.
- Application: the test material was introduced under a 2.5 cm × 2.5 cm cotton gauze patch and placed in position on the shorn skin.
- Type of wrap if used: the patch was secured in position with a strip of surgical adhesive tape. To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Dress removal: any residual test material was removed by gentle swabbing with cotton wool soaked in 74 % Industrial Methylated Spirits.
- Time after start of exposure: 4 hours.

OBSERVATION TIME POINTS
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation.

SCORING SYSTEM
The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the three test rabbits (12 values) and this total was divided by six to give the primary irritation index of the test material. The test material was classified according to the scheme devised by Draize J H (1959) "Dermal Toxicity" In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Association of Food and Drug Officials of the United States, Austin, Texas, p.47.

Primary Irritation Index // Classification of Irritancy
0: Non-irritant
> 0 to 2 Mild irritant
> 2 to 5 Moderate irritant
> 5 to 8 Severe irritant

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 eschar formation preventing grading of erythema 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 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible within: 48 hrs

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test material produced a primary irritation index of 0.2 and was classified as a mild irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Any other information on results incl. tables

Individual Skin Reactions

Animal N.	Reaction	1 hr	24 hrs	48 hrs	72 hrs	Mean 24/48/72 hrs
Male 152	Erythema	1 STA	0 STA	0 STA	0	0.00
Male 153	Erythema	1 STA	0 STA	0 STA	0 STA	0.00
Male 154	Erythema	1 STA	1 STA	0 STA	0	0.33
Male 152	Oedema	0	0	0	0	0.00
Male 153	Oedema	0	0	0	0	0.00
Male 154	Oedema	0	0	0	0	0.00
Sum of 24 and 72-hour Readings (S)		1
Primary Irritation Index (S/6)		1/6 = 0.2
Classification		Mild irritant

STA = Blue-coloured staining

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

Not skin irritant.

Executive summary:

The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method was designed to meet the requirements of the OECD guideline and the EU Method B4.

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced very slight erythema. Two treated skin sites appeared normal at the 24-hour observation and the remaining treated skin site appeared normal at the 48-hour observation. No signs of oedema were noted.

Conclusion

Mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions, thus the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC 1272/2008).