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EC number: 230-603-6
CAS number: 7216-56-0
7.3.1/1: Main test - Individual and mean OD values and tissue
viabilities for the test item, the negative and positive controls
Mean OD / disc (#)
Mean OD / product
Mean viability %
Standard deviation (SD)
mean of 3 values (triplicate of the same extract)
optical density; measured after a 1:2 dilution of the formazan extracts
SD ≤ 18%.
An in vitro skin irritation test
using the Reconstructed human Epidermis (SkinEthic RHE® model) was
performed according to Guideline OECD 439 and in compliance with GLP to
predict the acute skin irritation potential of the test substance.
The test substance was applied as supplied,
at the dose of 16 µL, to 3 living Reconstructed Human epidermis during
42 minutes, followed by a rinse with 25 mL of DPBS and a 41-hour and
8-minute post-incubation period at 37°C, 5% CO2. Cell viability was then
measured by enzymatic conversion of the vital dye MTT into a blue
formazan salt that was quantitatively measured after extraction from
In the preliminary tests, the test substance
was found not to have direct MTT reducing properties or colouring
potential. In the main test, the mean percent viability of the treated
tissues was 4.4%, versus 1.5% in the positive control (5% Sodium Dodecyl
Therefore, in accordance with Regulation EC
No 1272/2008, the test substance has to be classified in Category 2
“Irritating to skin”. The hazard statement “H315: Causes skin
irritation” with the signal word “Warning” are required.
Considering the results obtained, an in
vitro test of skin corrosivity has to be conducted to know if the
test item has to be classified in Category 1 “Corrosive” or Category 2
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