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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:
28th June 2012 to 8th November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Deviations:
no
Principles of method if other than guideline:
Not applicable.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): OLEIC_DimerFA_TETA_PAA
- Physical state: Yellow liquid with a brown hue.
- Analytical purity: 100%
- Lot/batch No.: BB100860V1
- Expiration date of the lot/batch: 30 April 2013
- Storage condition of test material: When not in use the test article was stored in a sealed container, at room temperature in the dark.

In vitro test system

Test system:
other: Three-dimensional human skin model
Source species:
human
Cell type:
other: Human keratinocytes
Cell source:
other: skin model EpiDermTM
Source strain:
not specified
Details on animal used as source of test system:
Not applicable
Justification for test system used:
The tissue has been shown to demonstrate reproducibility over time, and it has been shown to be capable of predicting the corrosive potential of the reference chemicals when using the testing protocol selected.
Vehicle:
unchanged (no vehicle)
Details on test system:
TEST SYSTEM

Specification
Three-dimensional human skin model, comprising a reconstructed epidermis with a functional stratum corneum supplied by MatTek Corporation, Ashland, Massachusetts, USA.

Identification
The test system was appropriately labelled with the study number, date of treatment, duration of treatment and negative/positive/test article.

General model conditions
Human keratinocytes are used to construct the epithelium. Multiple layers of viable epithelial cells are present under a functional stratum corneum. The stratum corneum is multi-layered with the necessary lipid profile to produce a functional barrier. The containment properties of the model prevent the passage of material around the stratum corneum to the viable model tissue. The skin model was supplied free of contamination with bacteria, mycoplasma and fungi.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
In order to assess the potential non-specific reduction of the test article, 40 mg of test article was added to 0.3 mL of 1.0 mg/mL MTT. In the Epiderm test, an average of approximately 44 mg of test article was applied to each tissue. Further tissues were concurrently treated with 40 μL distilled water (negative control) and with 40 μL 8N potassium hydroxide (positive control).
Duration of treatment / exposure:
3 minutes and 1 hour.
Duration of post-treatment incubation (if applicable):
Not applicable
Number of replicates:
duplicate

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean, after three minute exposure to the test article
Value:
72
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Not corrosive
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean, after one hour exposure to the test article
Value:
52
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Not corrosive

In vivo

Irritant / corrosive response data:
Skin viability after a three minute or one hour exposure to the test article was 72% and 52%, respectively.
Skin viability after a three minute or one hour exposure to the positive control article was 24% and 12%, respectively.
Other effects:
No other effects reported.

Any other information on results incl. tables

Three minute exposure period

Test substance

OD570

Mean

Tissue Mean

Adjusted tissue mean value

% variability

% survival

Negative

1.810

1.860

2.034

1.901

1.970

N/A

6.8

100

Negative

2.030

2.029

2.057

2.039

Test article

1.734

1.732

1.709

1.725

1.599

1.415

-17.1

72

Test article

1.445

1.446

1.530

1.474

Test article#

0.165

0.177

0.179

0.173

0.184

 

-12.7

Test article#

0.196

0.195

0.195

0.195

Positive

0.380

0.440

0.434

0.418

0.479

N/A

-29.071

24

Positive

0.542

0.531

0.545

0.539

# Freeze-killed tissues

One hour exposure period

Test substance

OD570

Mean

Tissue Mean

Adjusted tissue mean value

% variability

% survival

Negative

1.814

1.863

1.755

1.811

1.796

N/A

-1.6

100

Negative

1.700

1.832

1.814

1.782

Test article

1.147

1.237

1.215

1.200

1.238

0.926

6.0

52

Test article

1.262

1.339

1.2226

1.276

Test article#

0.262

0.274

0.273

0.270

0.312

 

-30.9

Test article#

0.351

0.354

0.355

0.354

Positive

0.238

0.224

0.236

0.233

0.222

N/A

9.271

12

Positive

0.212

0.221

0.201

0.211

# Freeze-killed tissues

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the test article, OLEIC_DimerFA_TETA_TEPA_PAA, was not considered to be corrosive to skin in the in vitro skin model EpiDerm.
Executive summary:

The skin corrosion potential of OLEIC_DimerFA_TETA_PAA was evaluated in a GLP study using the in vitro skin model EpiDermTM. The study was conducted in accordance with guidelines OECD Test Guideline 431 and EU Method B.40.

Duplicate EpiDermTM inserts were treated with OLEIC_DimerFA_TETA_PAA, distilled water (negative control) and 8N potassium hydroxide (positive control) for 3 minutes and 1 hour. At the end of the treatment period, the tissues were washed with phosphate buffered saline (PBS) and cell viability was assessed using the MTT assay. The skin corrosivity potential was classified according to the remaining cell viability obtained after test material treatment with either of the two treatment times.

Skin viability after a three minute or one hour exposure to the test article was 72% and 52%, respectively. Skin viability after a three minute or one hour exposure to the positive control article was 24% and 12%, respectively, demonstrating appropriate performance of the assay.

Under the conditions of this study, the test article, OLEIC_DimerFA_TETA_TEPA_PAA, was not considered to be corrosive to skin in the in vitro skin model EpiDermTM.