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EC number: 231-726-8
CAS number: 7704-98-5
The potential of the test item TiH2 to be corrosive to the skin was
investigated through an in vitro skin corrosion study, using a
commercial reconstructed human epidermis (RhE) model named EPISKIN™. The
experimental procedures are based on the OECD Guideline for testing of
chemicals no. 431. The test item, as well as controls, were tested for
their ability to impair cell viability after an exposure period of 3, 60
and 240 minutes. The final endpoint of the assay is the colorimetric
measurement of MTT reduction (blue formazan salt) in the test system,
being this reaction an index of cell viability. The test item was tested
as supplied by the Sponsor.
Before the Main Assay, a preliminary test was carried out to evaluate
the compatibility of the test item with the test system. In particular,
the test item was assayed for the ability of reducing MTT and colouring
water per se. After addition of the test item to water, a dark grey
solution was observed, with an OD value of 2.876, indicating a colouring
potential of the test item. Due to the dark colour of the substance, it
was not possible to clearly evaluate the actual ability of the test item
of reducing MTT. Based on these results, additional controls both for
colour potential and MTT unspecific reduction were added in the main
In the Main Assay, for each treatment time, the test item was applied as
supplied in two replicates at the treatment level of 20mg/epidermis
unit, each measuring 0.38cm2 (treatment level: 52.6 mg/cm2). Positive
and negative controls (Glacial acetic acid and Physiological saline,
respectively) were concurrently tested, in the same number of replicates
and test conditions at the treatment level of 50 μL/epidermis unit.
Positive control was included only at the longest treatment time of 240
minutes, while a negative control was included at each treatment time.
In theMain Assay, the negative controls gave the expected baseline value
(Optical Density values ≥ 0.6 and ≤ 1.5) and variability (difference of
viability between the two replicates lower than 30%), at each treatment
time, in agreement with the guideline indications. For each treatment
time, the concurrent negative control mean value is considered the
of the treatment series and thus represents 100% of cell viability.
The positive control caused the expected cell death (1% of cell
viability, when compared to the negative control).
Based on the stated criteria, the assay was regarded as valid.
Following treatment of alive tissue without MTT, colouring ability of
the test item was noted. For each treatment time, the calculated percent
value, above the concurrent negative control (NSCliving), was as follows:
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