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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2017-03-27 to 2017-04-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: MatTek Corporation Protocol: EpiOcular™ Eye Irritation Test (OCL-200-EIT) for the prediction of acute ocular irritation of chemicals; for use with MatTek Corporation’s Reconstructed Human EpiOcular™ Model
Version / remarks:
29 June 2015
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
Version / remarks:
July, 2015
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
3-hydroxypropyl octanoate
EC Number:
844-232-8
Cas Number:
102731-54-4
Molecular formula:
C11H22O3
IUPAC Name:
3-hydroxypropyl octanoate

Test animals / tissue source

Species:
human
Details on test animals or tissues and environmental conditions:
- Justification of the test method and considerations regarding applicability
In a prevalidation study performed by Avon Products Inc. and MatTek Corporation, the in vitro eye test using the human cornea model EpiOcular™ and measurement of cell viability by dehydrogenase conversion of MTT into a blue formazan salt have turned out as a sufficiently promising predictor for eye irritancy potential.
The EpiOcular ™ Eye Irritation Test (EIT) was validated by the European Union Reference laboratory for Alternatives to Animal Testing (EURL ECVAM) and cosmetics Europe between 2008 and 2013.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 50 μL
Duration of treatment / exposure:
30 min
Duration of post- treatment incubation (in vitro):
120 min
Number of animals or in vitro replicates:
2
Details on study design:
- Details of the test procedure used
- RhCE tissue construct used, including batch number: EpiOcular™ tissue, 23774
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods: standard culture conditions (37 ± 1.5 °C, 5 ± 0.5% CO2, 95% RH), only for 12 minute immersion incubation (post-soak) room temperature was used
- Indication of controls used for direct MTT-reducers and/or colouring test chemicals: Test item showed no direct MTT reducing or colouring properties. Therefore, no further controls were needed.
- Wavelength used for quantifying MTT formazan, and information on measuring device: The absorbance at 570 nm (OD570) of each well was measured with a plate reader (Versamax® Molecular Devices, 85737 Ismaning, Germany, Software Softmax Pro, version 4.7.1).
- Description of the method used to quantify MTT formazan: incubation in MTT solution for 180 min, extraction in isopropanol for ca 17 h at 2-8 °C and 2 h at RT with shaking
- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model:
If the test item-treated tissue viability is > 60 % relative to the negative control treated tissue viability, the test item is labeled non-irritant. If the test item-treated tissue viability is ≤ 60 % relative to negative control treated tissue viability, the test item is labeled irritant. A single test composed of at least two tissue replicates should be sufficient for a test chemical, when the result is unequivocal. However, in cases of borderline results, such as non-concordant replicate measurements and/or mean percent tissue viability equal to 60±5 %, a second test should be considered, as well as a third one in case of discordant results between the first two tests.
- Acceptability of assaya:
1) The negative control OD is > 0.8 and < 2.5,
2) The mean relative viability of the positive control is below 50% of the negative control viability.
3) The difference of viability between the two relating tissues of a single test item is < 20% in the same run (for positive and negative control tissues and tissues of test items).

Results and discussion

In vitro

Results
Irritation parameter:
other: cell viability [%]
Run / experiment:
mean
Value:
9
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100
Positive controls validity:
valid
Remarks:
40
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: no

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The negative control OD is > 0.8 and < 2.5 (1.922 and 2.068).
- Acceptance criteria met for positive control: The mean relative viability of the positive control is below 50 % of the negative control viability (40.0 %).
- The difference of viability between the two relating tissues of a single item is < 20 % (values between 0.6 % and 3.3 %) in the same run (for positive and negative control tissues and tissues of single test items).

Any other information on results incl. tables
























































































Dose
Group
Tissue
No.
Absorbance
570 nm
Well 1
Absorbance
570 nm
Well 2
Mean Absorbance
of
2 Wells
Mean-
Absorbance
of 2 Wells
Blank
corrected
Mean
Absorbance
of 2
Tissues
after Blank
Correction
Rel.
Absorbance
[%]
Tissue
1 + 2
Absolute Value of
the Difference of
the Rel.
Absorbances [%]
Tissue 1 and 2
Mean Rel.
Absorbance
[% of
Negative
Control]
Blank 0.0380.0390.0380    
Negative
Control
11.9282.0681.9981.961.927101.73.3100
21.9221.9461.9341.89598.3
Positive
Control
10.780.8490.8150.7760.77140.30.640
20.8020.8050.8030.76539.7
Test Item10.2260.2420.2340.1950.17310.12.39
20.1920.1890.190.1527.9

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The test item was possesses an eye irritating potential based on a study conducted according to OECD 492.
Executive summary:

An in vitro study according to OECD 492 was performed to assess the eye irritation potential of the test item by means of the Human Cornea Model Test. The test item did not prove to be an MTT reducer in the MTT pre-test, and it did not dye water or isopropanol in the colour interference pre-test. Therefore, additional tests with freeze-killed or viable tissues did not have to be performed. Each 50 μL of the test item, the negative control (deionised water) or the positive control (methyl acetate) were applied to each of duplicate tissue for 30 minutes. After treatment with the negative control the absorbance values were well within the required acceptability criterion of OD > 0.8 and < 2.5 thus showing the quality of the tissues. Treatment with the positive control induced a decrease below 50 % compared with the negative control value in the relative absorbance thus ensuring the validity of the test system. The difference of viability between the two relating tissues was < 20 % in the same run (for test item tissues, positive and negative control tissues). Irritating effects were observed following incubation with the test item. Compared with the value of the negative control the relative mean absorption value corresponding to the viability of the tissues decreased below 60 % (9.0 %). In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item possesses an eye irritating potential.

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