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EC number: 946-959-7
CAS number: -
Table 1: The first experiment: OD570 values obtained at the MTT test, their averages, standard deviations (%) and relative viabilities
5% SDS (PC)
Table 2: The second experiment: OD570 values obtained at the MTT test, their averages, standard deviations (%) and relative viabilities
negative, positive control
C3, C4, 394/16
standard deviation calculated from individual % tissue viabilities
viability of single tissues compared with negative control
damaged tissue (with lost of part of tissue)
damaged tissue (without lost of part of tissue)
The test substance, Acid Black 227, was assayed for in vitro skin irritation in the human epidermal model EpiDermTM. The test was performed according to the OECD Test Guideline No. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method (2015) and Protocol for: In Vitro EpiDermTM Skin Irritation Test For use with MatTek Corporation’s Reconstructed Human Epidermal Model EPI-200-SIT (see par. 1.4).
In the preliminary experiments neither colour interference with the endpoint nor direct MTT reduction were found.
After pre-incubation of tissues, 25 mg of the test substance was placed directly on previously moistened tissue and spread on the entire tissue surface. The length of exposure was 60 minutes. Three tissues were used for the test substance and for positive and negative controls.
After removal of the test substance, tissues were post-incubated for approximately 42 hours. Three hours incubation with MTT and two hours extraction period with shaking followed then. Optical density (OD570) of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues.
Under the above-described experimental design average viability of treated tissues was 36.9 % in the first and 25.8 % (after correction) in the second experiment, i.e. viability was < 50 %.
The effect of the test substance was positive in EpiDermTM model (tissues were damaged).
The test substance is identified as requiring classification and labelling according to UN GHS (Category 2 or Category 1). As the test substance was not tested for skin corrosion, it is not possible to decide between these two categories.
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