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

Endpoint summary

Description of key information

Based on the in vitro study results, the test substance is not considered to be irritating to skin and eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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)

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

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

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%).

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.

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).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 01, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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

Species:

cattle

Strain:

other: Bos primigenius Taurus

Details on test animals or tissues and environmental conditions:

Bovine corneas were used. They were collected from slaughtered cattle (from Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany, on the day of the test) which were between 12 and 60 months old.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 µL (the “closed chamber-method” was used in this study, for liquid substances)

Duration of treatment / exposure:

10 min at 32 ± 1 °C

Duration of post- treatment incubation (in vitro):

2 hours

Number of animals or in vitro replicates:

3

Details on study design:

The test substance was brought onto the cornea of a bovine eye which had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 h and whose opacity had been measured. The test substance was incubated on the cornea for 10 min at 32 ± 1 °C. After removal of the test substance and 2 h post-incubation, opacity and permeability values were measured

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea opacity and permeability

Value:

0.44

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:

Physiological sodium chloride solution was used as negative control and showed no irritating effect on the cornea and the calculated IVIS (in vitro irritancy score) was -0.79. Dimethylformamide (DMF) undiluted was used as positive control and induced serious eye damage on the cornea and fell within two standard deviations of the current historical mean. The calculated IVIS (in vitro irritancy score) was 96.29.
Under the conditions of this study, the test substance showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) was 0.44. According to OECD Guideline no. 437, a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage.

Interpretation of results:

other: CLP criteria not met

Remarks:

does not require classification

Conclusions:

Under the study conditions, the test substance was not irritating to bovine eye (BCOP test).

Executive summary:

A study was conducted to determine the in vitro eye irritation potential of the test substance according to OECD Guideline 437 and EU Method B.47 (BCOP test), in complinace with GLP. Bovine eyes were exposed to 750 µL undiluted test substance for 10 min (in triplicates). After removal of the test substance and 2 h post-incubation, opacity and permeability values were measured and reported as the in vitro irritancy score (IVIS). Physiological sodium chloride solution was used as negative control and dimethylformamide (DMF) undiluted as positive control. Physiological sodium chloride solution showed no irritating effect on the cornea and the calculated IVIS was -0.79. DMF induced serious eye damage on the cornea and fell within two standard deviations of the current historical mean. The calculated IVIS was 96.29. The test substance showed no effects on the cornea of the bovine eye. The calculated IVIS was 0.44. All validity criteria were met. Under the study conditions, the test substance was not irritating to bovine eye (Andres, 2016).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro skin irritation/corrosion:

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 irritating to the skin (Andres, 2016).

In vitro eye irritation:

A study was conducted to determine the in vitro eye irritation potential of the test substance according to OECD Guideline 437 and EU Method B.47 (BCOP test), in complinace with GLP. Bovine eyes were exposed to 750 µL undiluted test substance for 10 min (in triplicates). After removal of the test substance and 2 h post-incubation, opacity and permeability values were measured and reported as the in vitro irritancy score (IVIS). Physiological sodium chloride solution was used as negative control and dimethylformamide (DMF) undiluted as positive control. Physiological sodium chloride solution showed no irritating effect on the cornea and the calculated IVIS was -0.79. DMF induced serious eye damage on the cornea and fell within two standard deviations of the current historical mean. The calculated IVIS was 96.29. The test substance showed no effects on the cornea of the bovine eye. The calculated IVIS was 0.44. All validity criteria were met. Under the study conditions, the test substance was not irritating to bovine eye (Andres, 2016).

Justification for classification or non-classification

Based on the results of in vitro studies, the test substance does not need to be classified for skin and eye irritation according to CLP criteria (EC 1272/2008).