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EC number: 483-360-5
CAS number: 114435-02-8
Table 1: OD540 Values and %
viabilities for the negative control, positive control and test material
Mean % Viability4
1.895 ± 0.100
1.824 ± 0.071
0.258 ± 0.056
13.6 ± 3.0
0.118 ± 0.002
6.5 ± 0.1
1.583 ± 0.025
83.6 ± 1.3
1.205 ± 0.087
66.1 ± 4.7
= OD540 representing each EpiDerm tissue
= Mean OD540 of duplicate EpiDerm tissues ± standard deviation
= Viability representing each EpiDerm tissue expressed as a percentage
of the mean OD540 of either the 3 minute or 60 minute negative control
= Mean viability of duplicate treated EpiDerm tissues ± standard
= Mean viability of negative control tissues set at 100%
The study was performed to assess the
corrosivity potential of the test material using the EpiDerm Skin Model
(MatTek, Ashland, MA, USA). The test is based on the hypothesis that
corrosivity potential is related to toxicity to the EpiDerm tissue. The
study was validated by the inclusion of a positive control material, 8.0
N Potassium Hydroxide.
The experimental design of the study
consists of a test for Direct Reduction of MTT by the test material,
followed by the main test.
For the main test, duplicate EpiDerm tissues
were treated with 50 μL of Monofluoroethylene carbonate and exposed for
3 minutes and 60 minutes. The tissues were incubated at 37°C in a
humidified atmosphere of 5% C02 in air for the appropriate exposure
Duplicate negative and positive control
tissues, treated with 50 μL sterile distilled water or 50 μL 8.0 N
Potassium Hydroxide respectively, were also exposed for 3 minutes and 60
At the end of the exposure period each
EpiDerm tissue was rinsed using Dulbecco' s phosphate buffered saline
(DPBS) and placed into a 'holding plate', until all of the tissues had
been treated and rinsed. They were then transferred to an MTT 'loading
plate', and incubated at 37°C for 3 hours in a humidified atmosphere of
5% C02 in air. At the end of this time, each EpiDerm tissue was blotted
dry and placed into an MTT 'extraction plate' in order to extract all of
the reduced MTT from the tissues.
At the end of the extraction period, the
extracted MTT solution was mixed for each EpiDerm tissue and 3 x 200 μL
samples for each tissue were transferred to the appropriate wells of a
96 well plate. The absorbency at 540nm (OD540) of each well was measured
with the Anthos 2001 microplate reader. Data are presented in the form
of % viability (MTT conversion relative to negative controls) for each
of the two exposure times.
The relative mean viability of
Monofluoroethylene carbonate treated tissues was 83 .6% after 3 minutes
exposure and 66.1 % after 60 minutes exposure.
The quality criteria required for acceptance
of results in the test were satisfied.
Monofluoroethylene carbonate was considered
to be non-corrosive in vivo.
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