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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 15, 2019- May 17, 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Required by REACH

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report Date:
2019

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
July 29, 2016
Deviations:
no
Qualifier:
according to
Guideline:
other: Method B.40bis of Commission Regulation (EC) No 440/2008, of 30 May 2008, laying down test methods pursuant to Regulation (EC) No 1907/2006, 18 December 2006, of the European Parliament and of the Council on the REACH regulation
Version / remarks:
May 30, 2008
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
Purity: 99.7%
Physical description: white powder
Storage conditions: room temperature
Expiration date: March 20, 2020

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: The reconstructed human epidermal model EpiDerm™ (EPI-200, MatTek, Ashland, USA) consists of normal human-derived epidermal keratinocytes, which have been cultured to form a multilayered highly differentiated model of the human epidermis.
Source strain:
other: Human
Justification for test system used:
The information is required according to the REACH regulation Annex IIV section 8
Vehicle:
unchanged (no vehicle)
Details on test system:
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. Viable cells are able to reduce MTT to formazan whereas
non-viable cells cannot. The results are used to make a prediction of the corrosivity potential
of the test item (increased cytotoxicity is indicative of corrosion potential).

Methods:
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 directly reduce MTT and therefore additional non-viable tissues were
incorporated into the testing for correction purposes. 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 of 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).
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
The test item was used as supplied
Duration of treatment / exposure:
3 minutes
60 minutes
Duration of post-treatment incubation (if applicable):
Rinsing was achieved by filling and emptying each tissue under a constant soft stream of Dulbecco’s Phosphate Buffered Saline (DPBS) (without Ca++ Mg++) for approximately 40 seconds, to gently remove any residual test item. Excess DPBS was removed by blotting the bottom of
the tissue insert with tissue paper. Each tissue was placed into the prepared holding plate until all tissues were rinsed.
Number of replicates:
Duplicates

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
DOPO-HQ 3 minutes
Value:
>= 90.4
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
DOPO-HQ 60 minutes
Value:
>= 77.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Negative control 3 minutes
Value:
>= 100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Negative control 60 minutes
Value:
>= 100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Positive control 3 minutes
Value:
>= 4.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Positive control 60 minutes
Value:
>= 3.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
The criteria required for acceptance of results in the test were satisfied

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test item was considered to be non-corrosive to the skin.
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. Viable cells are able to reduce MTT to formazan whereas

non-viable cells cannot. The results are used to make a prediction of the corrosivity potential

of the test item (increased cytotoxicity is indicative of corrosion potential).

The test item was considered to be non-corrosive to the skin.