Benzenesulfonamide, N-[2-[[(phenylamino)carbonyl]amino]phenyl]-

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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

Test guideline 2

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:

1-(2-benzenesulfonamidophenyl)-3-phenylurea

EC Number:

806-543-7

Cas Number:

215917-77-4

Molecular formula:

C19H17N3O3S

IUPAC Name:

1-(2-benzenesulfonamidophenyl)-3-phenylurea

Test material form:

solid: particulate/powder

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Details on test system:

TEST SYSTEM
EPISKIN Small ModelTM (EPISKIN-SM(TM), 0.38 cm2, Batch no.: 14-EKIN-024)
Source: SkinEthic Laboratories, Lyon, France.
Rationale: One of the validated in vitro skin irritation tests is the EPISKIN test, which is recommended in international guidelines (e.g. OECD, EC and
validated by ECVAM).

TREATMENT
The test was performed on a total of 3 tissues per test substance together with negative and positive controls. The skin was moistened with 5 μl Milli-Q water (Millipore Corp., Bedford, Mass., USA) to ensure close contact of the test substance to the tissue and the solid test substance (11.5 to17.5 mg) was added into 12-well plates on top of the skin tissues. Three tissues were treated with 25 μl PBS (negative control) and 3 tissues with 25 μl 5% SDS (positive control) respectively. The positive control was re-spread after 7 minutes contact time. After the exposure period of 15 minutes at room temperature, the tissues were washed with phosphate buffered saline to remove residual test substance. After rinsing the cell culture inserts were each dried carefully and moved to a new well on 2 ml pre-warmed maintenance medium until all tissues were dosed and rinsed. Subsequently the skin tissues were incubated for 42 hours at 37°C.

CELL VIABILITY MEASUREMENT
Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues. Skin irritation potential of the test substance was classified according to remaining cell viability following exposure of the test substance.

All incubations, with the exception of the test substance incubation of 15 minutes at room temperature, were carried out in a controlled environment.
-Humid atmosphere of 80 - 100% (actual range 69 - 87%), containing 5.0 ± 0.5% CO2 in air in the dark.
-Temperature: 37.0 ± 1.0°C (actual range 36.0 – 36.9°C).
Temperature and humidity were continuously monitored throughout the experiment. The CO2 percentage was monitored once on each working day. Temporary deviations from the humidity were caused by opening of the incubator door. Based on HCD, these deviations were not considered to have affected the integrity of the study.

Control samples:

yes, concurrent vehicle

Amount/concentration applied:

11.5 to 17.5 mg

Duration of treatment / exposure:

15 minutes

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 times

Value:

98

Vehicle controls validity:

valid

Remarks:

100

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks:

11

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Mean Tissue Viability in the in vitro skin irritation test

	Mean tissue viability (percentage of control)
Negative control	100
Test substance	98
Positive control	11

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.