Fatty acids, tall-oil, reaction products with ethanolamine, ethoxylated

Endpoint summary

Description of key information

The results obtained from the in vitro skin irritation and corrosion test, using the EpiDerm model, indicated that the test item reveals no skin irritation potential under the utilised testing conditions. 
Application of the test item into the rabbits' eye mucosa caused slight to severe conjunctival and slight to moderate corneal irritant effects, fully reversible within 2 weeks.

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Two skin irritation tests and one eye irritation study are available for the substance to be registered (Fatty acids, tall-oil, reaction products with ethanolamine, ethoxylated (1,5 EO).

Skin Irritation

The potential of the test item to induce skin irritation / corrosion was first tested on reconstituted human epidermis in the EpiDerm model in vitro.

Skin corrosion in vitro:

Two EpiDerm™ tissue samples were incubated with the test substance for 3 minutes and 1 hour, respectively at room temperature in the incubator. Test groups consisted of test material, negative control and positive control; 12 tissues per test were used. The viability of each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37 °C. The resulting formazan chrystals were extracted with acidified isopropanol and quantified with the optical densities (OD) recorded spectrophotometrically. Positive and negative controls showed the expected cell viability values within acceptable limits. The experiment was considered to be valid. The results obtained from this in vitro skin corrosion test, using the EpiDerm model, indicated that the test item reveals no skin corrosion potential under the utilised testing conditions and is therefore not classified.

Skin irritation in vitro:

Disks of epidermal units (three units / chemical) were treated with the test item and incubated for 15 minutes at room temperature. Exposure of the test material was terminated by rinsing the epidermal units with PBS 1x solution. Epidermis units were then incubated at 37 °C for 42 hours. The viability of each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37 °C. The resulting formazan chrystals were extracted with acidified isopropanol and quantified with the optical densities (OD) recorded spectrophotometrically

SDS 5 % aq. and 1 x PBS treated epidermis units were used as positive and negative controls, respectively. For each treated tissue viability was expressed as a percentage relative to negative control.

In this in vitro skin irritation test using the EpiDerm model, the test item did not show significantly reduced cell viability in comparison to the negative control. All obtained test item viability results were above 50 % when compared to the viability values obtained from the negative control, therefore the test item was considered to be non-irritant to skin.

Positive and negative controls showed the expected cell viability values within acceptable limits. The experiment was considered to be valid.

The results obtained from this in vitro skin irritation test, using the EpiDerm model, indicated that the test item reveals no skin irritation potential under the utilised testing conditions and is therefore not classified.

Furthermore, an in vivo test was conducted with the test item to verify the in vitro test result. This seemed to be justified, as a Buehler test conducted with suspensions of the substance in water gave evidence for strong irritative or even corrosive effects after dermal application.

Skin corrosion/irritation in vivoo:

In a skin irritation/corrosion study according to OECD guideline No. 404, 0.5 mL of the test substance were applied on the clipped skin of New Zealand White rabbits (semi-occlusive patch, 4 hours). For animal welfare reasons, the test was started using a single animal and, as no severe skin lesions were observed in this animal, was continued with two further animals to establish a full scale study. The results of the three animals were pooled for evaluation.

In the first animal well-defined erythema (grade 2) and very slight edema (grade 1) were observed directly and 1 hour after removal of the patch. In the second animal well-defined erythema (grade 2) was noted directly after removal of the patch up to 24 hours after removal of the patch; at the 48 hours reading very slight erythema (grade 1) was observed. The third animal revealed well-defined erythema (grade 2) directly after removal of the patch up to 48 hours after removal of the patch; at the 72 hours reading very slight erythema (grade 1) was observed. Very slight edema (grade 1) was noted directly and 1 hour after removal of the patch in this aimal. The cutaneous reactions were reversible in one animal within 24 hours, in one animal within 72 hours and in the third animal within 7 days after removal of the patch. Mean scores over 24, 48 and 72 hours for each animal were 0.0, 1.0 and 1.7 for erythema and 0.0, 0.0 and 0.0 for edema. Under the test conditions chosen, the target substance was not corrosive or irritating to the skin.

 

Eye irritation

The acute eye irritation study of the test item was performed in three female Littlerussian rabbits. The irritation effect of the test item was evaluated according to the Draize method (OECD No.: 405, 2002). The test item was placed into the conjunctival sac of the left eye of each animal. The untreated right eye served as control. A weight of 0.1 mL of the test item was used in pure state, as a single dose. The eyes of the test animals were not washed out after the application of test item. The eyes were examined at 1, 24, 48 and 72 hour(s) and then at 1 and 2 weeks after the application.

One hour after application, a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light were observed in all animals. A diffuse, crimson red conjunctiva with individual vessels not easily discernible, a swelling above normal and a discharge with moistening of the lids and hairs just adjacent to lids were observed in two animals and a discharge different from normal in animal 3. 24 hours after application of the test article a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible, an obvious swelling with partial eversion of lids and a discharge with moistening of the lids and hairs, and considerable area around the eye were observed in all animals. After instillation of Fluorescein all animals showed a scattered or diffuse area of opacity on more than half, but less than three quarters of the cornea. 48 hours after application of the test article after instillation of Fluorescein two animals showed a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, whereas a scattered or diffuse area of opacity on one quarter or less of the cornea was observed in other animal. 72 hours after application of the test article two animals showed a scattered or diffuse area of opacity on more than quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as a swelling above normal. 7 days after application of the test article some conjunctival vessels definitely injected were observed in two animals No. 1 and No. 2. Animal No.3 did not show any signs of eye irritation. 14 days after application of the test article animals No. 1 and No. 2 were free of any signs of eye irritation, too.

The individual mean cornea scores of 24, 48 and 72 hours were 1,1 and 0.67 for animal No. 1,2 and 3, respectively. The individual mean iris score for all animals is 1. Individual mean conjunctivae scores were determined to 2, 2 and 2.67 and the individual mean chemosis scores were 1.33, 2 and 2 in animal No. 1, 2 and 3, respectively.

In conclusion, test item applied to the rabbits' eye mucosa, caused slight to severe conjunctival and slight to moderate corneal irritant effects, fully reversible within 2 weeks.

Justification for selection of skin irritation / corrosion endpoint:
In vivo study according to GLP and OECD guidelines.

Justification for selection of eye irritation endpoint:
In vivo study according to GLP and OECD/EU guidelines.

Effects on eye irritation: irritating

Justification for classification or non-classification

Based on the available experimental data the test item is classified as eye irritating Category 2 (H319), Causes serious eye irritation according to Regulation (EC) 1272/2008 (CLP) and as Xi, R36 Irritanting to the eyes according to Directive 67/548/EC (DSD). No classification according to Regulation (EC) No 1272/2008 and Directive 67/548/EEC is required with respect to skin irritation/corrosion is required based on the experimental data available.