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EC number: 947-964-7
CAS number: -
OD values and viability percentages of the controls:
Optical Density (OD)
Negative Control:Sterile deionized water
Positive Control:Methyl acetate
OD values and viability percentages of the test item:
standard deviation (SD)
study according to OECD Guideline 492 was conducted to determine the
acute ocular irritation potential of the test item on three-dimensional
RhCE tissue in the EpiOcular™ model in vitro. Before treatment the
tissues were pre-wetted with approximately 20 μL of Ca++Mg++Free-DPBS
and were incubated at standard culture conditions for 30 ± 2 minutes.
Disks of EpiOcular™ (two units) were treated with test item (50
μL/units) and incubated for 30 ± 2 minutes at standard culture
conditions (37±2°C in an incubator with 5±1% CO2, ≥95% humidified
atmosphere). Exposure of test material was terminated by rinsing with
Ca++Mg++ Free-DPBS solution. After rinsing, the tissues were incubated
for a 12 ± 2 minutes immersion incubation (Post-Soak) at room
temperature. At the end of the Post-Soak immersion test items treated
tissues were incubated for 120 ± 15 minutes at standard culture
conditions (Post-treatment Incubation). Fresh assay medium was used
during the Post-Soak and Post-incubation. The viability of each disk was
assessed by incubating the tissues for 3 hours (±15 min) with MTT
solution at 37±2°C in an incubator with 5±1% CO2 protected from light,
≥95% humidified atmosphere. The formazan precipitated was then extracted
using isopropanol and quantified spectrophotometrically. Sterile
deionized water and methyl acetate treated tissues were used as negative
and positive controls respectively. The Disks of EpiOcular™ (two units /
control) were treated with positive and negative control and incubated
30 ± 2 minutes. The test item did not show significantly reduced cell
viability in comparison to the negative control (mean viability: 93 %).
All obtained test item viability results were above 60 % when compared
to the viability values obtained from the negative control. Therefore
the test item was considered to be non-irritant to the eye. Positive and
negative controls showed the expected cell viability values within
acceptable limits. The experiment was considered to be valid.
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