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EC number: 237-354-2
CAS number: 13760-80-0
Table 2: relative mean viabilities for each treatment group
*The mean viability of the negative control tissues is set at 100%
Table 3: Mean OD562 Values and Viabilities for the Negative Control,
Positive Control and Test Material
Mean OD562 of individual tissues
Mean OD562 of duplicate tissues
Coefficient of Variation
Relative Mean Viability (%)
Relative mean % tissue viability = (mean OD562 of test material / mean
OD562 of negative control) x 100
Coefficient of variation = (standard deviation / mean OD562 of duplicate
tissues) x 100
The potential of the test material to cause skin corrosion was
determined in accordance with the standardised guidelines OECD 431 and
EU method B40.bis, under GLP conditions using the EpiDerm Human Skin
Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue. Cytotoxicity
is determined by the reduction of MTT
(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to
formazan by viable cells in the test material treated tissues relative
to the corresponding negative control. The results are
used to make a prediction of the corrosivity potential of the test
Before the main test was performed the test material was tested for
direct MTT Reduction, the MTT solution containing the test material did
not turn blue/purple. This was taken to indicate the
test material did not reduce MTT. The test material was also assessed
for its potential to cause colour interference with the MTT endpoint.
The solution containing the test material did not become coloured. This
was taken to indicate the test material did not have the potential to
cause colour interference.
Duplicate tissues were treated with the test material for exposure
periods of 3 and 60 minutes. Negative and positive
control groups were treated for each exposure period. At
the end of the exposure period the test material was rinsed from each
tissue before each tissue was taken for MTT loading. After
MTT loading each tissue was placed in 2 mL isopropanol for MTT
At the end of the formazan extraction period each well was mixed
thoroughly and triplicate 200 µL samples were transferred to the
appropriate wells of a pre-labelled 96 well plate. The
optical density (OD) was measured at 562 nm (OD562).
The negative control, positive control and coefficient of variation were
all within the acceptability criteria and so the study was valid.
Under the conditions of the study the test material was considered to be
non-corrosive to the skin.
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