Disodium 2,4-diamino-3-{[4-(ethenylsulfonyl)phenyl]diazenyl}-5-{[4-(ethenylsulfonyl)-2-sulfonatophenyl]diazenyl}benzenesulfonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

Principles of method if other than guideline:

Guidelines followed

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

None

Test animals

Species:

other: skin tissue

Strain:

not specified

Details on test animals or test system and environmental conditions:

Test system:

EpiDerm Skin Model (EPI-200, Lot no.: 20532 kit N).

The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.

Test system

Type of coverage:

other: EpiDerm Skin Model

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

not required

Amount / concentration applied:

None

Duration of treatment / exposure:

None

Observation period:

Acceptability of the assay:
The in vitro skin corrosion test is considered acceptable if it meets the following criteria:
a) The absolute mean OD540 of the two tissues of the negative control should reasonably be within the laboratory historical control data range.
b) The mean relative tissue viability following 3-minute exposure to the positive control should be <= 30%.
c) The maximum inter-tissue variability (in viability) is 30% between two tissues treated identically.
d) The maximum difference in percentage between the mean viability of two tissues and one of the
two tissues is 15%.

Number of animals:

None

Details on study design:

None

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1-hour application viability (percentage of control)

Value:

92

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3-minute application viability (percentage of control)

Value:

88

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Irritant / corrosive response data:

Finally, it is concluded that this test is valid and that FAT 40866/A TE is not corrosive in the in vitro skin corrosion test under the experimental conditions.

Other effects:

None

Any other information on results incl. tables

FAT 40866/A TE was checked for possible direct MTT reduction by adding the test substance to MTT medium. Because no colour change was observed it was concluded that FAT 40866/A TE did not interact with MTT.

The mean absorption at 540 nm measured after treatment with FAT 40866/A TE.

The mean tissue viability obtained after 3-minute and 1-hour treatments with FAT 40866/A TE compared to the negative control tissues. Skin corrosion is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after the 3-minute and 1-hour treatments with FAT 40866/A TE compared to the negative control tissues was

88% and 92% respectively. Because the mean relative tissue viability for FAT 40866/A TE was not below 50% after 3 minutes treatment and not below 15% after 1 hour treatment FAT 40866/A TE is considered to be not corrosive.

The absolute mean OD540 (optical density at 540 nm) of the negative control tissues was within the laboratory historical control data range. The mean relative tissue viability following 3-minute exposure to the positive control was 9%. The maximum inter-tissue variability in viability between two tissues treated identically was less than 20% and the maximum difference in percentage

between the mean viability of two tissues and one of the two tissues was less than 11%. It was therefore concluded that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:

other: not corrosive

Remarks:

Criteria used for interpretation of results: OECD GHS

Conclusions:

Finally, it is concluded that this test is valid and that FAT 40866/A TE is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.

Executive summary:

In vitro skin corrosion test with FAT 40866/A TE using a human skin model. This report describes the ability of FAT 40866/A TE to induce skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential of FAT 40866/A TE was tested through topical application for 3 minutes and 1 hour.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch BOP 04-13 (Lot: BS-MIF 975) of FAT 40866/A TE was a red-brown powder. Skin tissue was moistened with 25 μl of Milli-Q water and 25 mg of FAT 40866/A TE was applied directly on top of the skin tissue.

The positive control had a mean relative tissue viability of 9% after 3 minutes exposure. The absolute mean OD540 (optical density at 540 nm) of the negative control tissues was within the laboratory historical control data range. The maximum inter-tissue variability in viability between two tissues treated identically was less than 20% and the maximum difference in percentage between the mean

viability of two tissues and one of the two tissues was less than 11%, indicating that the test system functioned properly.

Skin corrosion is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with FAT 40866/A TE compared to the negative control tissues was 88% and 92%, respectively. Because the mean relative tissue viability for FAT 40866/A TE was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment FAT 40866/A TE is considered to be not corrosive.

Finally, it is concluded that this test is valid and that FAT 40866/A TE is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.