Mono Dimer Trimer FA TETA PAA

Skin irritation / corrosion

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

Test guideline 2

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

Test material information

Constituent 1

Reference substance name:

High molecular weight adducts of Fatty acids, C18-unsatd dimers and trimers with amines, polyethylenepoly-, triethylenetetramine fraction

IUPAC Name:

High molecular weight adducts of Fatty acids, C18-unsatd dimers and trimers with amines, polyethylenepoly-, triethylenetetramine fraction

Constituent 2

Reference substance name:

Low Molecular weight adducts of Fatty acids, C16-18 sat. C18 unsat., linear, dimers, with Amines, polyethylenepoly-, triethylenetetramine fraction

IUPAC Name:

Low Molecular weight adducts of Fatty acids, C16-18 sat. C18 unsat., linear, dimers, with Amines, polyethylenepoly-, triethylenetetramine fraction

Constituent 3

Reference substance name:

Amines, polyethylenepoly-, triethylenetetramine fraction

EC Number:

292-588-2

EC Name:

Amines, polyethylenepoly-, triethylenetetramine fraction

Cas Number:

90640-67-8

Molecular formula:

C6H18N4, C8H20N4, C8H20N4, C6H18N4

IUPAC Name:

Amines, polyethylenepoly-, triethylenetetramine fraction

Details on test material:

- Name of test material (as cited in study report):OLEIC DimerFA TETA PAA
- Physical state:Yellow-red, transparent, viscous liquid
- Analytical purity: 8% free amine
- Storage condition of test material:Room temperature (15°C to 30°C)
-Sponsor batch: BB100860V1

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

Results

Results 1

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

Results 2

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

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 EpiDerm^TM. The study was conducted in accordance with guidelines OECD Test Guideline 431 and EU Method B.40.

Duplicate EpiDerm^TM 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 EpiDerm^TM.