Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
other: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
January from 21st to 22nd, 2009
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
The reliability of the source read-across study was established to be R1: guideline study
Justification for type of information:
Justification for read-across is detailed at section 13.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
April 2004
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: three-dimensional RhE model (EpiDermTM).
- Souce: Mattek (USA)
- Tissue batch number: lot 10810/A.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during exposure: 37 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: 20 washings using PBS.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- Reagent: 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl tetrazolium bromide (MTT).
- MTT concentration: 300 µl/plate.
- Incubation time: 3 hours.
- Extraction: isopropanol (2000 µl).
- Wavelength: 550 nm.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
20 mg of test item per unit of epidermis.
Duration of treatment / exposure:
3 minutes and 60 minutes (at 37 °C, 5 % CO2)
Duration of post-treatment incubation (if applicable):
3 hours
Number of replicates:
Test material and controls (positive and negative) were applied on each unit of epidermis in duplicate.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Time point: 3 minutes
Value:
ca. 105.1
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
Standard deviation: 7.5.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Time point: 60 minutes
Value:
ca. 122.5
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
Standard deviation: 26.6.
Other effects / acceptance of results:
Classification: non corrosive

Any other information on results incl. tables

RESULTS

SAMPLE

Mean cell viability (SD)-3 minutes exposure

Mean cell viability (SD)-60 minutes exposure

Classification

SET RETARD PLUS

105.1 (7.5)

122.5 (26.6)

NON CORROSIVE

KOH 8N

6.0 (0.6)

5.8 (1.1)

CORROSIVE

NEGATIVE CONTROL

100.0 (4.8)

100.0 (2.0)

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Applicant's summary and conclusion

Interpretation of results:
other: non corrosive, i.e. the substance does not meet the criteria to be classified as Skin Corr. 1A/B/C, according to the CLP Regulation
Conclusions:
Non corrosive.
Executive summary:

Method

Corrosion characteristics of test item were evaluated according to the guideline OECD 431 (April 13, 2004), on reconstitued epidermis 3D (EpiDerm™), through MTT test cytotoxicity analysis.

The test item was applied on each epidermis unit in duplicate for 3 minutes and 1 hour at 37°C, 5% CO2. At the end of the exposure, the test item was removed, and cell viability evaluated through MTT test.

Observations

The percentage of mean cell viability for treated cells resulted to be:

- after 3 minutes: 105.1 %

- after 60 minutes: 122.5 %

Results

Non corrosive, i.e. the substance does not meet the criteria to be classified as Skin Corr. 1A/B/C, according to the CLP Regulation