Registration Dossier

Diss Factsheets

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 guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
N-{2-[(phenylcarbamoyl)amino]phenyl}benzenesulfonamide
EC Number:
806-543-7
Cas Number:
215917-77-4
Molecular formula:
C19H17N3O3S
IUPAC Name:
N-{2-[(phenylcarbamoyl)amino]phenyl}benzenesulfonamide
Specific details on test material used for the study:
Batch: CDC-003

Test animals

Species:
other: EPISKIN Small model (Human)
Details on test animals or test system and environmental conditions:
ENVIRONMENTAL CONDITIONS
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.

Test system

Vehicle:
physiological saline
Amount / concentration applied:
11.5 to 17.5 mg
Duration of treatment / exposure:
15 minutes
Details on study design:
TEST SYSTEM
Name: 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 to
17.5 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.

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.