Registration Dossier

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:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
skin obtained from plastic surgery from multiple donors
Source strain:
Wistar
Details on animal used as source of test system:
The test system is a commercially available EpiDermTM-Kit, procured by MatTek.
The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, 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 representing main
lipid classes analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell culture inserts.
Vehicle:
water
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Number of replicates:
3

Test animals

Species:
rat
Strain:
Wistar

Test system

Type of coverage:
semiocclusive
Preparation of test site:
abraded
Vehicle:
water
Controls:
yes, concurrent positive control
yes, concurrent negative control
Duration of treatment / exposure:
One plate (3 tissues) was used as negative control; each tissue was treated with 30 µL DPBS buffer, a nylon mesh was added in order to ensure sufficient contact with the tissue surface.
One plate was used as positive control; each tissue was treated with 30 µL 5% SDS-solution, a nylon mesh was added in order to ensure sufficient contact with the tissue surface.
One plate was used for treatment with the test item:
The tissues were wetted with 25 µL DPBS buffer before applying the test item and spread-ing it to match the tissue size.

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
99.7
Vehicle controls validity:
valid
Negative controls validity:
not valid
Positive controls validity:
valid

Any other information on results incl. tables

% Tissue Viability

Designation

N-(4-anilinophenyl)-4-
methylbenzenesulfonamide

Positive
Control

% Tissue viability (tissue 1)

98.6%

2.1%

% Tissue viability (tissue 2)

99.7%

2.2%

% Tissue viability (tissue 3)

101.5%

2.4%

% Tissue viability (mean)

99.9%

2.2%

± SD of mean tissue viability (%)

1.5%

0.2%

        Validity

Criterion

Demanded

Found

OD of negative control

≥0.8 and≤ 2.8

1.5

% tissue viability
of positive control SDS

£20% of negative control

2.2% %

SD of mean viability of the

tissue replicates (%)

≤18%

1.6% (negative control)
0.2% (positive control)
1.5% (test item)

 

Two experiments were performed.

The first experiment was not valid, because the standard deviation of the three replicates of the test item was 18.5%, required is ≤18%. This experiment is not reported in this report, but the raw data are kept in the GLP- archive of the test facility.

After the treatment, the mean value of relative tissue viability was reduced to 99.9%. This value is above the threshold for skin irritation (50%).

The test item N-(4-anilinophenyl)-4-methylbenzenesulfonamide is considered as non-irritant to skin.

The optical density of the negative control was well within the required acceptability crite-rion of 0.8 ≤ mean OD ≤ 2.8.

The positive control has met the acceptance criterion too, for thus ensuring the validity of the test system.

Variation within replicates was within the accepted range for negative control, positive control and test item (required: ≤ 18%).

For these reasons, the result of the test is considered valid.

Applicant's summary and conclusion

Conclusions:
The test item N-(4-anilinophenyl)-4-methylbenzenesulfonamide is considered as non-irritant to skin.