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EC number: 500-215-4
CAS number: 68475-94-5
Mean OD570 Values and Viabilities for the Negative Control Item,
Positive Control Item and Test Item
Mean OD570of triplicate tissues
Relative individual tissue viability (%)
Relative mean viability (%)
± SD of Relative mean viability (%)
Negative Control Item
Positive Control Item
OD = Optical Density
SD = Standard deviation
* = The mean viability of the negative control tissues is set
The purpose of this test was to evaluate the skin irritation potential
of the test item using the EPISKINTM reconstructed human epidermis model
after a treatment period of 15 minutes followed by a post-exposure
incubation period of 42 hours. The principle of the assay was based on
the measurement of cytotoxicity in reconstructed human epidermal
cultures following topical exposure to the test item by means of the
colorimetric MTT reduction assay. Cell viability is measured by
enzymatic reduction of the yellow MTT tetrazolium salt
(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a
blue/purple formazan salt (within the mitochondria of viable cells) in
the test item treated tissues relative to the negative controls.
Triplicate tissues were treated with the test item for an exposure
period of 15 minutes. At the end of the exposure period each tissue was
rinsed before incubating for 42 hours. At the end of the post-exposure
incubation period each tissue was taken for MTT-loading. The maintenance
medium from beneath each tissue was transferred to pre-labelled micro
tubes and stored in a freezer for possible inflammatory mediator
determination. After MTT-loading a total biopsy of each epidermis was
made and placed into micro tubes containing acidified isopropanol for
extraction of formazan crystals out of the MTT-loaded tissues.
At the end of the formazan extraction period each tube was mixed
thoroughly and duplicate 200 μL samples were transferred to the
appropriate wells of a pre-labelled 96-well plate. The optical density
was measured at 570 nm.
Data are presented in the form of percentage viability (MTT reduction in
the test item treated tissues relative to negative control tissues).
The relative mean viability of the test item treated tissues was 93.3%
after the 15-Minute exposure period and 42-Hours post-exposure
Quality criteria: The quality criteria required for acceptance of
results in the test were satisfied.
The test item was classified as non-irritant. The following
classification criteria apply:
EU CLP Not classified for Irritation.
UN GHS Not classified for Irritation (category 3 cannot be determined).
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