Registration Dossier

Administrative data

Description of key information

According to the results of the key studies, the test item 2 -Amino-4 -Hydroxyethylaminoanisole sulfate (A084) was not considered as Skin Irritant substance. Hence, the test item is not classfifed for Skin Irritation.

Two key studies were available for Eye Irritation, according the results of his studies, the test item was classified as Category 2A for Eye Irritation.

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)

Additional information

Two key studies were available for Skin irritation :

Busching, 1979, OECD 404, Klimisch 2, GLP compliant

10 female albino guinea pigs were used. The test substance was aplied to an area of 3 x 4 am of the right and left flanks of the animals with a brush and was washed off after 20 minutes. The treatment was repeated 3 times on two consecutive days.The treatment period was followed by a 3-day observation period. 2 of 10 animals showed a very slight erythema of the clipped not sacrificied skin. There were no edemas observed. On the last day of the experiment, all of the animals were free from symptoms. Behaviour of the animals was not changed during the observation period. Thus, the test item was not considered as a skin irritant.

Warren, 2018, OECD 439, Klimisch 1, GLP compliant

Triplicate tissues of EPISKINTM model were treated with the test item for an exposure period of 15 minutes.  At the end of the exposure period each tissue was rinsed before incubating for 42 hours. DPBS was used as negative control and SDS as positive control condition. The test item was found to directly reduce MTT and therefore additional non-viable tissues were incorporated into the testing for correction purposes.  At the end of the post exposure incubation period each tissue was taken for MTT-loading. The relative mean viability of the test item treated tissues was 86.1% after the 15 Minute exposure period and 42 Hours post exposure incubation period.

Two key studies were availables for Eye Irritation Assessment:

-The purpose of the first study (Prinsen, 2016, OECD 438, GLP compliant, Klimisch 1) was to evaluate the potential irritation to eye property of the test item A084 when applied neat in chicken eye .Chicken eyes were obtained from slaughter animals used for human consumption. The isolated chicken eyes were exposed to a single application of 30 mg for 10 seconds followed by a 20 mL saline rinse. Three main parameters were measured to disclose possible adverse eye effects: corneal thickness (expressed as corneal swelling), corneal opacity and fluorescein retention of damaged epithelial cells. In addition, histopathology of the corneas was performed. 2-Amino-4-Hydroxyethylaminoanisole Sulfate caused slight corneal swelling (mean swelling 9%), moderate opacity (mean score 2.0) and moderate fluorescein retention (mean score 2.0). Microscopic examination of the corneas revealed very slight (1/3 corneas) erosion of the epithelium.Hence, the test item cannot be precisely classified for Eye Irritation.

-This in vitro study (Duschl, 2017, GLP compliant, OECD 492, Klimisch 1) was performed to assess the eye irritation potential of 2-Amino-4-Hydroxyethylaminoanisole sulfate (A084) by means of the Human Cornea Model Test. The test item proved to be an MTT reducer in the MTT pre-test, and it changed colour (slightly reddish) in the presence of water in the colour interference pre-test. Therefore, additional tests with freeze-killed and with viable tissues (without MTT addition) had to be performed to determine correction factors for calculating the true viability in the main experiment.

Each 50 mg of the test item, were applied to each of duplicate tissue for 6 hours. Each 50 µL of the negative control (deionised water) and of the positive control (methyl acetate) were also applied to duplicate tissues each. Viability of the tissues were quantified using MTT reduction approach. Irritating effects were observed following incubation with 2-Amino-4-Hydroxyethylaminoanisole sulfate (A084). Compared with the value of the negative control the relative mean absorption corresponding to the viability of the tissues decreased to 13.0% (threshold for irritancy: < 60%). In conclusion, it can be stated that in this study and under the experimental conditions reported, 2-Amino-4-Hydroxyethylaminoanisole sulfate (A084) possesses an eye irritating potential.

Justification for classification or non-classification

For Skin Irritation, based on the key study results and according to the CLP criteria, the test item 2 -Hydroxyethylaminoanisole sulfate (A084) was not classified for Skin Irritation.

For Eye Irritation, based on the key studies result and according to the CLP criteria, the test item was classified as Category 2A for Eye Irritation. In the first in vitro ICE study, the test item was not considered as Not Irritating substance but induced damages to epithelial cells which were not sufficient to be considered as corrosive substance. Hence, a second in vitro study was performed. In a Human Cornea Model Test (OECD 492), the test item showed eye irritating potential. According the results of the two studies and by caution, the test item showed Eye Irritation Potential and was classified as Category 2A for Eye Irritation.