4,4'-ethylenedianiline

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well performed GLP study according to OECD 439

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 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-ethylenedianiline

EC Number:

210-716-7

EC Name:

4,4'-ethylenedianiline

Cas Number:

621-95-4

Molecular formula:

C14H16N2

IUPAC Name:

4-[2-(4-aminophenyl)ethyl]aniline

Test animals

Species:

other: reconstituted human epidermis model

Strain:

other: reconstituted human epidermis model

Test system

Type of coverage:

other: Topical

Preparation of test site:

other: Not applicable

Vehicle:

other: Dulbecco's Phosphate Buffered Saline (DPBS)

Amount / concentration applied:

Each approximately 25 mg of the test item were applied to the tissues, wetted with 25 µL Dulbecco's Phosphate Buffered Saline (DPBS), and spread to match the surface of the tissue.
VEHICLE: Dulbecco's Phosphate Buffered Saline (DPBS)

Duration of treatment / exposure:

60 minutes

Observation period:

Not applicable

Number of animals:

Not applicable

Details on study design:

Each three tissues of the human skin model EpiDerm™ were treated with the test item, the negative or the positive control for 60 minutes.
Approximately 25 mg of the test item were applied to each tissue, wetted with 25 µL of DPBS, and spread to match the surface of triplicate tissue.
30 µL of either the negative control (DPBS) or the positive control (5% SLS) were applied to triplicate tissue each.
The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 43 hours the tissues were treated with the MTT solution for 3 hours following 69.75 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: % Relative Absorbance

Run / experiment:

1

Value:

66.6

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

other: % Relative Absorbance

Run / experiment:

2

Value:

68.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation / corrosion parameter:

other: % Relative Absorbance

Run / experiment:

3

Value:

75.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 4

Irritation / corrosion parameter:

other: % Relative Absorbance

Run / experiment:

mean

Value:

70

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Results after treatment with DABY, trocken and the controls :

Dose Group	Treat-ment Interval	Absor-bance 570 nm Tissue 1*	Absor-bance 570 nm Tissue 2*	Absor-bance 570 nm Tissue 3*	Mean Absor-bance of 3 Tissues	Rel. Absor-bance [%] Tissue 1, 2, and 3**	Relative Standard Deviation [%]	Mean Rel. Absorbance [% of Negative Control]***
Negative Control	60 min	1.742	1.808	1.904	1.818	95.8 99.5 104.7	4.5	100.0
Positive Control	60 min	0.126	0.117	0.103	0.115	6.9 6.4 5.7	9.7	6.3
Test Item	60 min	1.211	1.241	1.367	1.273	66.6 68.2 75.2	6.5	70.0

*       Mean of three replicate wells after blank correction

**       relative absorbance per tissue [rounded values]: (100 x (absorbance tissue)) / (mean absorbance negative control)

***     relative absorbance per treatment group [rounded values]: (100 x (mean absorbance test item / positive control)) / (mean absorbance negative control)

 

Since the test item was not coloured, there was no need to perform the test for colour interference.

Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour.

The mean relative absorbance value of the test item, corresponding to the cell viability, decreased to 70.0% (threshold for irritancy: ≤ 50%), consequently the test item was not irritant to skin.

 

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, DABY, trocken is not irritant to skin according to UN GHS and EU CLP regulation.

Executive summary:

This in vitro study was performed to assess the irritation potential of DABY, trocken by means of the Human Skin Model Test.

Each three tissues of the human skin model EpiDerm^™were treated with the test item, the negative or the positive control for 60 minutes.

Approximately 25 mg of the test item were applied to each tissue, wetted with 25 µL of DPBS, and spread to match the surface of triplicate tissue.

30 µL of either the negative control (DPBS) or the positive control (5% SLS) were applied to triplicate tissue each.

The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 43 hours the tissues were treated with the MTT solution for 3 hours following 69.75 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD ≥0.8 and ≤ 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.

Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 6.3% thus ensuring the validity of the test system.

The rel. standard deviations between the % variabilities of the test item, the positive and negative controls were below 10% (threshold of the "OECD Guideline for the Testing of Chemicals 439:In vitroSkin Irritation: Reconstructed Human Epidermis Test Method”: 18%), thus ensuring the validity of the study.

Compared to the relative absorbance value of the negative control the mean relative absorbance value was reduced to 70.0% after exposure of the skin tissues to the test item. This value is above the threshold for irritancy of ≤ 50%. Therefore, the test item is not considered to possess an irritant potential.