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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation / corrosion
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27th June 2012 to 12th December 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted according to current guidelines.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other 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
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
Qualifier:
according to guideline
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

Constituent 1
Reference substance name:
DimerFA_PEPA_PAA
IUPAC Name:
DimerFA_PEPA_PAA
Constituent 2
Reference substance name:
Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines
EC Number:
614-452-7
Cas Number:
68410-23-1
IUPAC Name:
Fatty acids, C18-unsatd., dimers, reaction products with polyethylenepolyamines
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): DimerFA_PEPA_PAA
- Physical state: yellow/brown liquid
- Analytical purity: 100%
- Lot/batch No.: BB 001694 V1
- Expiration date of the lot/batch: 31st August 2013
- Storage condition of test material: When not in use the test article was stored in a sealed container, at 15 to 25°C in the dark.

Test animals

Species:
other: Not applicable - in vitro study
Strain:
other: Not applicable - in vitro study
Details on test animals or test system and environmental conditions:
Not applicable - in vitro study

Test system

Type of coverage:
other: Not applicable
Preparation of test site:
other: Not applicable
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
In order to assess the potential non-specific reduction of the test article, the test article was added to 0.3 mL of 1.0 mg/mL MTT.
In the EpiDerm test, an average of approximately 27 mg of test article was applied to each tissue.
Duration of treatment / exposure:
3 minutes and 1 hour.
Observation period:
Not applicable.
Number of animals:
Not applicable
Details on study design:
On the day of receipt EpiDermTM tissues were placed in a refrigerator. The next day, at least one hour before starting the assay, the tissues were transferred to 6-well plates with the assay medium, which was replaced immediately before the test was started.
The test was performed on a total of four tissues per test article, negative and positive control, out of which two were used for a 3 minute application and two tissues were used for a 1 hour application. Sufficient test article was applied to cover the outer layer of the tissue. The cryovials containing the test article were weighed both prior to and post treatment and an average of approximately 27 mg of test article was applied to each tissue.
Further tissues were concurrently treated with 40 μL purified water (negative control) and with 40 μL 8N potassium hydroxide (positive control). After the 3-minute or 1-hour contact periods, the tissues were washed with phosphate buffered saline (PBS) to remove residual material. The rinsed tissues were kept in 24-well plates (holding plates) until all tissues were dosed and rinsed.

Once all tissues had been rinsed, they were transferred to wells containing 300 μL of 1 mg/mL MTT-medium and were incubated for 3 hours (37°C). After incubation, the tissues were washed with PBS and any resultant colour was extracted with 2 mL isopropanol overnight. The optical density of each resultant extract was determined spectrophotometrically at 570 nm and cell viability was calculated for each tissue as a percentage of the mean of the negative control tissue. Skin corrosivity potential of the test article was classified according to the remaining cell viability obtained after test article treatment with either of the two treatment times.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: Skin viability
Basis:
mean
Time point:
other: 3 minute exposure
Remarks on result:
other: 90% skin viability
Irritation parameter:
other: Skin viability
Basis:
mean
Time point:
other: 1 hour exposure
Remarks on result:
other: 64% skin viability
Irritant / corrosive response data:
Skin viability after a three minute or one hour exposure to the test article was 90% and 64%, respectively.
Skin viability after a three minute or one hour exposure to the positive control article was 27% and 13%, respectively.
Other effects:
No additional information.

Any other information on results incl. tables

Three minute exposure

Test substance

OD570

Mean

Tissue Mean

Adjusted mean value

% variability

% survival

Negative

0.641

0.690

0.706

0.679

0.955

N/A

44.9

100

Negative

1.224

1.217

1.254

1.232

Test article

0.914

0.938

1.006

0.953

1.065

0.857

19.2

90

Test article

1.189

1.215

1.130

1.178

Test article#

0.231

0.231

0.232

0.231

0.208

 

20.0

Test article#

0.187

0.183

0.185

0.185

Positive

0.298

0.310

0.318

0.309

0.260

N/A

31.4

27

Positive

0.212

0.212

0.212

0.212

# Freeze-killed tissues

 

One hour exposure

Test substance

OD570

Mean

Tissue Mean

Adjusted mean value

% variability

% survival

Negative

1.298

1.357

0.1360

1.338

1.375

N/A/

4.8

100

Negative

1.404

1.414

1.399

1.406

Test article

1.061

0.961

0.903

0.975

1.076

0.880

17.1

64

Test article

1.279

1.171

1.079

1.176

Test article#

0.184

0.183

0.183

0.183

0.195

 

-12.8

Test article#

0.205

0.210

0.206

0.207

Positive

0.156

0.155

0.154

0.155

0.173

N/A

-23.3

13

Positive

0.190

0.195

0.187

0.191

# 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 and based on the results obtained, DimerFA_PEPA_PAA, was not considered to be corrosive to the skin using the in
vitro skin model: EpiDerm.
Executive summary:

A study to determine the potential of DimerFA_PEPA_PAA to cause skin corrosion was conducted using the in vitro skin model: EpiDermTM. The study was conducted in accordance with OECD Test Guideline 431 and EU Method B.40 and was compliant with GLP.

Duplicate EpiDermTM inserts were treated with DimerFA_PEPA_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 assessed 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 90% and 64%, respectively. Skin viability after a three minute or one hour exposure to the positive control article was 27% and 13%, respectively, demonstrating appropriate performance of the assay.

Under the conditions of this study and based on the results obtained, DimerFA_PEPA_PAA, was considered not to be corrosive to skin in the in vitro skin model EpiDermTM.