2-((E)-(4-((4-(1,3-dioxo-1,3-dihydro-2H-hetereocarbapolycyclo-2-yl)alkyl)(ethyl)amino)-2-methyl)diazenyl)-5-nitro-1,3-dicarbonitrile, polyphenyl

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008-09-18 till 2008-09 19

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for Testing of Chemicals 431: In vitro Skin Corrosion: Human Skin Model Test (Original Guideline adopted April 13, 2004).

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: "Council Regulation (EC) No 44012008 B40 bis", dated 30 May, 2008.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of the test material (as cited in the study report: FAT 40842/A TE
- Substance type: coloring dye
- Physical state: solid, dark bluish green powder
- Analytical purity: 96%
- Lot/batch No.: Blau DRI 2098 Op 1/07
- Expiration date of the lot/batch: June 30, 2014
- Storage condition of test material: at room temperature at about 20 ºC

Test animals

Species:

other: In Vitro Test: Human Skin Model

Strain:

other: not applicable

Details on test animals or test system and environmental conditions:

not applicable: In Vitro Test: Human Skin Model

Test system

Type of coverage:

other: not applicable: In Vitro Test: Human Skin Model

Preparation of test site:

other: not applicable: In Vitro Test: Human Skin Model

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25mg / tissue

(VEHICLE)
- Amount(s) applied (volume or weight with unit): About 25 mg of the test material was wetted with 50 μL deionised water.

Duration of treatment / exposure:

2 treatment intervals: 3 minutes and 1 hour

Observation period:

not applicable: Human Skin Model Test: consists of topical application of the test material to the tissue for 3 minutes and 1 hour, followed by immediate determination of the cytotoxic effect.

Number of animals:

not applicable: In Vitro Test: Human Skin Model

Details on study design:

The test allows the discrimination between corrosive and non-corrosive chemical substances and mixtures. It does not provide information on skin irritation, nor does it allow the sub-categorisation of corrosive substances according to GHS.

The In vitro Skin Corrosion: Human Skin Model Test consists of topical application of the test material to the tissue for 3 minutes and 1 hour, followed by immediate determination of the cytotoxic effect. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the exposure period.

Duplicate EpiDerm™ tissues were exposed to the test item, positive control or negative control for each of two different exposure periods: 3 minutes and 1 hour.

At the end of the exposure period the tissues were removed from the 6-well plate and gently rinsed using a wash bottle containing PBS to remove any residual test material. Excess PBS was removed by gently shaking the tissue insert and blotting the lower surface with blotting paper. The tissues were placed in the prepared holding plate until all tissues were rinsed.

After the exposure procedure was completed for all tissues of each time point, the cell culture inserts were transferred from the holding plates to the MTT-plates for the MTT Assay.

Results and discussion

In vivo

Irritant / corrosive response data:

In the present study, the test item FAT 40842/A TE was tested for its potential to induce skin corrosion in a human skin model. The test allows the discrimination between corrosive and non-corrosive chemical substances and mixtures. It does not provide information on skin irritation, nor does it allow the sub-categorisation of corrosive
substances according to GHS.

Any other information on results incl. tables

Results after Treatment with FAT 40842/A TE

Dose group	Treat-ment Interval	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Mean Absorbance of 2 Tissues	Rel. Absorbance [% of Negative Control]**
Negative Control	3 min	1.658	1.859	1.759	100.0
Positive Control	3 min	0.318	0.338	0.328	18.7
FAT 40842/A TE	3 min	1.753	1.334	1.544	87.8
Negative Control	1 hour	1.549	1.751	1.650	100.0
Positive Control	1 hour	0.178	0.174	0.176	10.7
FAT 40842/A TE	1 hour	1.643	1.645	1.644	99.6

* Mean of three replicate wells after blank correction

** relative absorbance [rounded values]: 100 x (absorbance_testitem ) / (absorbance_negative control

Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show evidence of a blue color and thereby was not considered to be an MTT reducer

Applicant's summary and conclusion

Interpretation of results:

other: not corrosive

Remarks:

Criteria used for interpretation of results: other: according to guideline, see also "Any other Information on materials and methods incl. tables"

Conclusions:

In conclusion, it can be stated that in this study and under the reported experimental conditions, the test item FAT 40842/A TE was non corrosive to skin.

Executive summary:

This in vitro study was performed to assess the corrosive potential of FAT 40842/A TE by means of the Human Skin Model Test.

Independent duplicate tissues of the human skin model EpiDerm™ were exposed to either the test item, the negative control or the positive control for 3 minutes and 1 hour, respectively.

Approximately 25 mg of the test item were spread to cover the surface of the tissue and wetted with 50 μL deionised water.

A volume of 50 μL of either the negative control (deionised water) or the positive control (8.0 N KOH) was applied to each tissue.

After exposure to the negative control the absorbance values exceeded the required acceptability criterion of mean OD570 ≥ 0.8 for both treatment intervals thereby confirming the acceptable quality of the tissues.

Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 minutes exposure period and for the 1 hour exposure period thus confirming the validity of the test system.

After exposure to the test item FAT 40842/A TE the relative absorbance values were irrelevantly decreased to 87.8% after 3 minutes. After the 1 hour exposure relative absorbance values were not reduced (99.6%). Both values did not exceed the threshold for corrosivity (50% for the 3 minutes exposure and above 15% for the 1 hour exposure).

Therefore, the test item was not considered to be corrosive.