Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine

Endpoint summary

Description of key information

in vivo skin irritation/corrosion (OECD 404): irritating (Bioassay, 2013)
in vitro eye corrosion (OECD 437; BCOP): no risk of serious eye damage (BASF SE, 2013)
in vitro eye irritiation (EpiOcular): irritating (BASF SE, 2013)

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Skin irritation/corrosion:

The potential of Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine to cause acute dermal irritation or corrosion was assessed by a single topical application of an amount of 0.5 mL of the test item for 4 hours to the intact skin of three White New

Zealand rabbits (stepwise procedure starting with one animal and supplementing one additional animal per step up to a maximum number of three animals), using a patch of 2.5 cm x 2.5 cm, covered with semi-occlusive dressing. The study was conducted according to OECD 404 guideline and GLP (Bioassay, 2013). After removal of the patch the application area was washed off.

The cutaneous reactions were assessed immediately after removal of the patch, approximately 1, 24, 48 and 72 hours after removal of the patch and in weekly intervals until day 14.

The following test item-related clinical observations were recorded during the course of the study:

· Very slight to moderate erythema (grade 1-3)

· Very slight to severe edema (grade 1-4)

· erythema and edema beyond the application site

· In addition crusty secretion (from 24 until 72 hours), formation of plate-like incrustation (from 72 hours until day 7), scaling (from 24 hours until day 14), yellowish or brown-yellowish discoloration of the application area (from 24 until 72 hours) and petechiae (in one animal at day 7) were observed during the course of the study.

The cutaneous reactions were not reversible in all three animals within 14 days after removal of the patch. Erythema and/or edema were still present at study termination.

Necrosis of the skin was not evident during the course of the study.

Mean scores over 24, 48 and 72 hours for each animal were 3.0, 3.0 and 2.7 for erythema and 4.0, 4.0 and 4.0 for edema.

Considering the described cutaneous reactions as well as the average score for irritation, Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine shows a skin irritating potential under the test conditions chosen.

Eye irritation

EpiOcular

In a GLP compliant study the potential of Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamineto cause ocular irritation was assessed by a single topical application of 50μL of the undiluted test substance to a reconstructed three dimensional human cornea model (EpiOcular™)(BASF SE, 2013). Two EpiOcular™ tissue samples were incubated with the test substance for 30 minutes followed by a 2-hours post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability. The EpiOcular™ eye irritation test showed the following results: The test substance is able to reduce MTT directly. Therefore an additional MTT reduction control (KC) was introduced. As compound residues remained on the tissues after the washing procedure, the results of the MTT reduction control of the test substance indicated an increased MTT reduction (difference to KC of NC is greater than 0.1). Thus the KC was used for viability calculation. The mean viability of the test-substance treated tissues was 38% (Mean tissue viability (% of negative control) Prediction: ≤ 50 irritant; > 50 ≤ 60 no prediction; > 60 non-irritant). Based on the observed results and applying the evaluation criteria it was concluded, thatOleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine shows an eye irritation potentialin the EpiOcular™ eye irritation test under the test conditions chosen.

BCOP

In a GLP compliant study performed according to OECD guideline 437 the potential of Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamineto cause serious damage to the eyes was assessed by a single topical application of 750μL of the undiluted test substance to the epithelial surface of isolated bovine corneas (BASF SE, 2013). Three corneas were treated with the test substance for 10 minutes followed by a 2-hours post-incubation period. Corneal opacity was measured quantitatively as the amount of light transmission through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were used to calculate anIn VitroIrritancy Score of the test substance relative to the control corneas. The BCOP test showed the following results for the test substance: Mean Opacity Value: 3.4,Mean Permeability Value: 0.452, In Vitro Irritancy Score:10.2 (IVIS > 55 risk of serious eye damage, IVIS <= 55 no risk of serious eye damage).

In addition, minimal amounts of compound residues remained on the surface of the corneastreated with the test substance after the washing procedure. The mean opacity value of the negative control (NC) is minimal out of the historical range. However, as all other acceptance criteria were met in the test and due to the unambiguous results of the test substance, the evaluation of the study is notexpected to be influenced by this deviation. Based on the observed results and applying the evaluation criteria it was concluded, thatOleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine does not cause serious eye damagein the Bovine Corneal Opacity and Permeability Test (BCOP Test) under the test conditions chosen.

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

Based on the available data and according to Directive 67/548/EEC (DSD) the test item is classified as irritating to the skin and labelled with Xi and the risk phrase R38 "Irritating to the skin". According to the Regulation (EC) No 1272/2008 (CLP), the substance is classified as category 2 and labelled with the signal word "Warning" and H315 "causes skin irritation"

According to the harmonised classification criteria laid down in Council Directive 67/548/EEC, Annex VI (and subsequent adaptations) the test substance should be classified as irritating to the eyes and assigned the symbol Xi, and the risk phrase R36: "Irritating to eyes" should be applied. According to Annex I, Regulation (EC) No 1272/2008 (CLP), the substance is classified as category 2 and labelled with "Warning" and H319 "causes serious eye irritation".