Fatty acids, linseed-oil, reaction products with bisphenol A-epichlorohydrin polymer and glycidyl neodecanoate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 23, 2016 to March 11, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of oxiran-2-ylmethyl 2,2,3,5-tetramethylhexanoate with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:1)

Molecular formula:

Not applicable (UVCB constituent)

IUPAC Name:

Reaction product of oxiran-2-ylmethyl 2,2,3,5-tetramethylhexanoate with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:1)

Constituent 2

Reference substance name:

Reaction product of 4,4'-Methylendiphenyldiglycidylether (BADGE) with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:2)

Molecular formula:

Not applicable (UVCB constituent)

IUPAC Name:

Reaction product of 4,4'-Methylendiphenyldiglycidylether (BADGE) with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:2)

Constituent 3

Reference substance name:

Reaction product of 4,4'-Methylendiphenyldiglycidylether (BADGE) with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:1)

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Reaction product of 4,4'-Methylendiphenyldiglycidylether (BADGE) with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:1)

Constituent 4

Reference substance name:

Reaction product of 4,4'-Methylendiphenyldiglycidylether (BADGE) with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:3)

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Reaction product of 4,4'-Methylendiphenyldiglycidylether (BADGE) with linseed oil fatty acids (including majorly linolenic acid, linoleic acid and oleic acid) (1:3)

Constituent 5

Reference substance name:

High Mw components based on linseed oil fatty acids

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

High Mw components based on linseed oil fatty acids

Constituent 6

Reference substance name:

Sum of unassigned components

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Sum of unassigned components

Test material form:

liquid

Details on test material:

UVCB

Specific details on test material used for the study:

Batch no.: #210162718
Composition: reaction products of linseed-oil fatty acids, 4,4'Methylendiphenyldiglycidylether with neodecanoic fatty acid, oxiranylmethylester
Purity: 100 % as per definition of UVCB
Appearance: brown liquid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Details on animal used as source of test system:

Commercially available EpiDermTM-Kit (from MatTek In Vitro Life Science Laboratories, Bratislava). The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell cultures inserts.

Vehicle:

unchanged (no vehicle)

Details on test system:

The test consists of a topical exposure of the neat test substance to a human reconstructed epidermis model followed by a cell viability test. Cell viability is measured by dehydrogenase conversion of MTT 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide, present in cell mitochondria, into a blue formazan salt that is quantitatively measured after extraction from tissues. The percentage reduction of cell viability in comparison of untreated negative controls is used to predict skin irritation potential.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

30 µL of undissolved test substance

Duration of treatment / exposure:

60 min

Duration of post-treatment incubation (if applicable):

ca. 42h30. Following this post-incubation period, the MTT test was performed

Number of replicates:

3 tissues, 2 replicates

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Remarks:

% formazan production

Run / experiment:

MMT test (relative absorbance value)

Value:

98.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

After treatment with the negative control, the absorbance values were within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8, OD was 2.0. The positive control showed clear irritating effects. Relative absorbance was reduced to 3.2% (requirement: < 20%). Variation within tissues was acceptable (requirement: ≤18%). After the treatment with the test substance, the relative absorbance values were reduced to 98.5%. This value is well above the threshold for irritation potential (50%).

Any other information on results incl. tables

The first experiment was not valid, because one validity criterion of the negative control was not fulfilled. The standard deviation within the tissue replicates was too high. These data are not reported, but kept together with the raw data in the GLP archive of the test facility. The second experiment was valid.

Applicant's summary and conclusion

Interpretation of results:

other: CLP criteria not met

Remarks:

does not require classification

Conclusions:

Under the study conditions, the test substance was considered not to be irritant to the skin in the Human Skin Model Test.

Executive summary:

A study was conducted to determine the in vitro skin irritiation potential of the test substance according to OECD Guideline 439 and EU Method B.46 (Reconstructed Human Epidermis Test Method), in compliance with GLP. Three tissues were exposed for 60 min to 30 µL of undissolved test substance. After a post-incubation period of ca. 42h 30 min, the MTT test was performed. Indeed, cell viability was measured by dehydrogenase conversion of MTT, present in cell mitochondria, into a blue formazan salt that is quantitatively measured after extraction from tissues. The percentage reduction of cell viability in comparison of untreated negative controls was used to predict skin irritation potential. After treatment with the negative control (DPBS-buffer), the absorbance values were within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8, OD was 2.0. The positive control (sodium dodecyl sulphate) showed clear irritating effects. Relative absorbance was reduced to 3.2% (requirement: ≤18 %). After the treatment with the test substance, the relative absorbance values were reduced to 98.5%. This value is well above the threshold for irritation potential (50%). Based on the results obtained, the experiment was considered valid. Under the study conditions, the test substance was considered not to be irritant to the skin (Andres, 2016).