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EC number: 700-042-6
CAS number: 177997-13-6
The mean OD540 values
and mean viabilities for each treatment group are given in Table 1.
The relative mean viability of the test material treated tissues
after a 10 minute exposure was 7.0%.
It was considered unnecessary to proceed with tissue
The qualitative evaluation of tissue viability is presented in
The test material treated tissues appeared white and were
considered to be indicative of dead tissue. The negative control treated
tissues appeared blue and were considered to be indicative of viable
tissue. The positive control material treated tissues appeared
blue/white and were considered to be indicative of semi‑viable tissue.
Table1 Assessment of Eye Irritation Potential –
Viability of RHC Tissues
* = The mean viability
of the negative tissues is set at 100%
Table2 Qualitative Evaluation of Tissue Viability
(MTT uptake visual evaluation)
MTT Visual Scoring Scheme of
- = Blue
+ = Blue/White
tissue (semi viable)
++ = Tissue
completely white (dead)
The purpose of this study was to determine the eye irritation
potential of the test material using the SkinEthic Reconstituted Human
Corneal model (HCE) after a treatment period of 10 minutes. The test is
based on the hypothesis that irritant chemicals are able to penetrate
the corneal epithelial tissue and are sufficiently cytotoxic to cause
The experimental design of the study consists of a test for Direct
Reduction of MTT by the test materials, followed by the main test.
For the main test, triplicate SkinEthic tissues were treated with
30 mg of the test material for 10 minutes. Triplicate tissues treated
with 30 µl of Solution A served as the negative control and triplicate
tissues treated with 30 µl of 1% w/v Sodium Dodecyl Sulphate served as
the positive control.
At the end of the exposure period each SkinEthic tissue was
rinsed. The rinsed tissues (two per group) were taken for MTT
loading. The remaining tissues were retained for possible
histopathology. Following MTT loading the reduced MTT was extracted from
After extraction the absorbency of triplicate aliquots of the
extracted MTT solution for each SkinEthic tissue was measured (OD540). Data
are presented in the form of % viability (MTT conversion relative to
The test material was classified according to the following
the % relative mean tissue viability was ≥ 60% the test material was
considered to not likely to be a severe ocular irritant.
the % relative mean tissue viability was < 60% the test material was
considered to be irritant.
The quality criteria required for acceptance of results in the
test were satisfied.
Under the conditions of the test the test material was considered
to be an irritant (I). However, a subsequent in vivo skin
irritation test identified the test material as being corrosive to the
skin. Therefore the corrosive results of the in vivo skin
irritation test should also be taken into consideration when concluding
the eye irritation potential of the test material.
According to the protocol followed the
test material was considered to be Irritant (I).
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