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EC number: 221-361-2 | CAS number: 3077-27-8
In vitro skin corrosion, Reconstructed human epidermis test method (OECD 431): not corrosive
In vitro skin irriation, Reconstructed human epidermis test method (OECD 439): not irritating
In vitro eye irritation, Reconstructed human cornea-line epithelium model (OECD 492): not irritating
Table 1: Absorbance values blank isopropanol (OD 570nm)
Table 2: Absorbance Values (OD 570nm) for negative control, test item and positive control
Table 3: Mean Absorbance Values of the 3 min Experiment
Mean – blank (tissue 1)
Mean – blank (tissue 2)
Mean of the two tissues
Table: 4: Mean Absorbance Values of the 1 h Experiment
Table 5: % Tissue Viability
Table 6: Historical Data
Optical Density Negative Control
Optical Density Negative Con- trol
% Tissue viability Positive Control
1.197 - 3.077
1.377 - 2.571
9.6 - 57.3 %
4.1 - 24.2 %
Table 1: Absorbance Values negative control, test item and positive control (OD 570 nm)
(octadecylamino)-4- oxo-2-butenoic acid
Table 2: Mean Absorbance Values
Mean – blank (tissue 3)
Mean of the three tissues
Table 3: % Tissue Viability
(2Z)-4-(octadecylamino)- 4-oxo-2-butenoic acid
% Tissue viability (tissue 1)
% Tissue viability (tissue 2)
% Tissue viability (tissue 3)
% Tissue viability (mean)
± SD of mean tissue viability (%)
Measured optical densities and calculated values are summarised in the a separate pdf document (OECD 492 results.pdf)
In vitro skin corrosion:
The skin corrosion potential of the test substance was determined by an in vitro skin corrosion test using a reconstructed human skin model according to OECD guideline 431 and in compliance with GLP (LAUS, 2017a). Skin corrosion potential was identified by reduction of cell viability measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that is quantitatively measured after extraction from tissues. In this study cell viability after treatment with the test substance was 99.9% and 107.4% after 3 and 60 minutes, respectively. The acceptance criteria were met for the positive and negative control. Therefore, 4-(octadecylamino)-4-oxoisocrotonic acid is not considered to be corrosive to the skin.
In vitro skin irritation:
The skin irritation potential of the test substance was determined by an in vitro skin irritation test using a reconstructed human skin model according to OECD guideline 439 and in compliance with GLP (LAUS, 2017b). Skin irritation potential was identified by reduction of cell viability measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that is quantitatively measured after extraction from tissues. In this study, the mean cell viability of three tissues after treatment with the test substance was 100.9%. The acceptance criteria were met for the positive and negative control. In this study the test substance did not show irritating properties towards human-derived epidermal keratinocytes. Therefore, 4-(octadecylamino)-4-oxoisocrotonic acid is not considered to be a skin irritant.
In vitro eye irritation:
The eye irritation potential of the test substance was investigated in an in vitro study using a reconstructed human cornea-like epithelium according to OECD guideline 492 and observing GLP conditions (LAUS, 2018a). The test item was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 6 h. After treatment, the substance was rinsed from the tissue. Then the cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan formation was evaluated by measuring the optical density (OD) of the resulting solution. As the test item showed intense coloring in the pre-test, there was the risk to influence the photometric measurement. Therefore, an additional test for intensely coloured test items was performed. Demineralised water was used as negative control and methyl acetate was used as positive control. The controls showed the expected results. After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD > 0.8 and < 2.5 (OD was 2.0). The positive control showed clear eye irritating effects as the mean value of the relative tissue viability was 38.2 % (< 50%). Variation within tissue replicates of the controls and the test item was acceptable (< 20%). After treatment with the test item, the mean value of relative tissue viability was 103.7 %. This value is well above the threshold for identifying an eye irritation potential (≤ 60%). Therefore, the test item is considered not to be irritating to eyes.
The available data on skin irritation / corrosion and eye irritation of the registered substance do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 and are, therefore, conclusive but not sufficient for classification.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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