Reaction mass of 6-amino-2-anilino-4-methyl-5-{(E)-[4-nitro-2-(substituted)phenyl]diazenyl}nicotinonitrile and 2-amino-6-anilino-4-methyl-5-{(E)-[4-nitro-2-(substituted)phenyl]diazenyl}heteromonocyclic nitrile

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31-Oct-2011 to 04-Nov-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Guideline no. 431: In Vitro Skin Corrosion: Human Skin Model Test

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU method B.40 BIS: "In Vitro Skin Corrosion: Human Skin Model Test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test system

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg, moistened with 25 µl water

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 50 µl Milli-Q water

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µl KOH
- Concentration (if solution): 8 N

Duration of treatment / exposure:

Exposure:3 minutes and 1 hour

Details on study design:

TEST SITE
- Area of exposure: human skin model
- % coverage: 0.6 cm²

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 3 minutes and 1 hour


SCORING SYSTEM:
- Percentage viability: After treatment, the medium was replaced by 300 µl MTT-medium and tissues were incubated for
3 hours at 37 °C in air containing 5 % CO2. After incubation the tissues were washed with PBS and formazan was extracted with 2 ml isopropanol over night at room temperature. The amount of extracted formazan was determined spectrophotometrically at 540 nm.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: percentage viability

Value:

98

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 3 minutes. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: percentage viability

Value:

111

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 1 hour. (migrated information)

Applicant's summary and conclusion

Interpretation of results:

other: not corrosive

Conclusions:

The in vitro skin irritation test was conducted according to OECD 431 guideline and GLP principles. It is concluded that this test is valid and that FAT 41043/A is not corrosive in the in vitro skin irritation test.

Executive summary:

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 41043/A compared to the negative control tissues was 98 % and 111 %, respectively. Because the mean relative tissue viability for FAT 41043/A was not below 50 % after the 3-minute treatment and not below 15 % after the 1-hour treatment FAT 41043/A is considered to be not corrosive. The positive and negative controls induced appropriate responses.