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Skin irritation / corrosion

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Administrative data

skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 February 2018 - 12 March 2018
1 (reliable without restriction)

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations were considered to have not affected the integrity or validity of the study.
GLP compliance:

Test material

Constituent 1
Test material form:
semi-solid (amorphous): gel

In vitro test system

Test system:
isolated skin discs
Source species:
Cell type:
non-transformed keratinocytes
Cell source:
foreskin from multiple donors
Source strain:
Justification for test system used:
System used to supplant in-vivo testing
unchanged (no vehicle)
Details on test system:
Test System
EpiDerm™ Reconstructed Human Epidermis Model Kit

Supplier: MatTek
Date received: 22 February 2018
EpiDermTM Tissues (0.63cm2) lot number :25882
Assay Medium lot number:021518TMA
Upon receipt of the EpidermTM tissues, the sealed 24-well plate was stored in a refrigerator until use.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
50 uL
Duration of treatment / exposure:
3 min/60 min
Duration of post-treatment incubation (if applicable):
Number of replicates:
2 ea. for postive and negative controls and substance at 3min, 60 min.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1/ @3 minutes
ca. 97.8
Vehicle controls validity:
Negative controls validity:
Positive controls validity:
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
2/@ 60 min
ca. 111.8
Vehicle controls validity:
Negative controls validity:
Positive controls validity:

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Predicted to be non-corrosive to the skin via employed test system. See attached background material.
Executive summary:


The purpose of this test is to evaluate the corrosivity potential of the test item using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes.

Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue. Cytotoxicity is determined by the reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to formazan by viable cells in the test item treated tissues relative to the corresponding negative control. The results are used to make a prediction of the corrosivity potential of the test item.


Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes. Negative and positive control groups were treated for each exposure period. The test item was found to have the potential to cause color interference and therefore additional tissues were incorporated for color correction purposes. The test item was also found to directly reduce MTT and therefore, an additional procedure using freeze-killed tissues was performed. A third set of controls was included; comprising freeze-killed tissues, in order to prevent a double correction from a colored test item that also reduces MTT. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT-loading. After MTT loading each tissue was placed in 2 mL Isopropanol for MTT extraction.

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 μL samples were transferred to the appropriate wells of a pre-labeled 96-well plate. The optical density (OD) was measured at 570 nm (OD570).

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues). See attached background material.