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EC number: 277-616-3
CAS number: 73816-74-7
The test item, Reactive Red 45:1, was assayed for the in vitro
in human epidermal model EpiDermTM.The test was performed
according to the OECD Test Guideline No.439: In VitroSkin Irritation:
Reconstructed Human Epidermis Test Method (2015) and Protocol for: In
Vitro EpiDermTMSkin Irritation Test For use with MatTek
Corporation’s Reconstructed Human Epidermal Model EPI-200-SIT (see par.
After pre-incubation of tissues, 25 mg of the test item was placed
directly a top to the previously moistened tissue and it was spread on
the entire tissuesurface. Length of exposition was 60 minutes. Three
tissues were used for the test item and every control.
After removal of the test substance, tissues were post-incubated
42 hours due to leave of damage reparation. 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.
At direct reduction assay in a test tube MTT medium was coloured
red, so direct MTT reduction was excluded.
In the first experiment too high differenceamong viability of
tissues was observed in range close to cut off value. The experiment was
then repeated. In the repeated experiment all test conditions were
fulfilled and average viability of treated tissues was54.3% (53. 4 %
after correction),i.e. viability was >50 %.
The effect of the test item was negative in EpiDermTMmodel
(tissues were not damaged).
According to the classification criteria given in chapter 4.5.,
the test item, Reactive Red 45:1, is considered to have no category in
regard to skin irritation.
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