Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-02-20 to 2013-03-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to GLP, OECD 437, EU Method B. 47 and other similar study guidelines without deviations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
other: The Ocular Toxicity Working Group (OTWG) of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the NICEATM, The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
other: Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Trantos
- Molecular formula (if other than submission substance): C26H36O4S
- Molecular weight (if other than submission substance): 444.63
- Substance type: Pure active substance
- Physical state: solid, white powder
- Analytical purity: 99.9%
- Lot/batch No.: CQE299K1DL2
- Expiration date of the lot/batch: 15 January 2014
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark.
- Other: pH (1% in water, indicative range): 8.7-8.2

Test animals / tissue source

Species:
other: in vitro
Details on test animals or tissues and environmental conditions:
Test System: Bovine eyes were used as soon as possible after slaughter on the same day.

Source: Bovine eyes from young cattle were obtained from the slaughterhouse, where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter.

Transport: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):The medium from the anterior compartment was removed and 750 μl of the negative control and 20% (w/v) Imidazole solution (positive control) were introduced onto the epithelium of the cornea. TRANTOS was weighed in a bottle and applied directly on the corneas in such a way that the cornea was completely covered (314 to 374 mg). Corneas were incubated in a horizontal position for 240 ± 10 minutes at 32 ± 1°C.
Duration of treatment / exposure:
240 minutes
Number of animals or in vitro replicates:
9 eyes total were used for the study; 3 negative control tests, 3 positive control tests, and 3 tested with Trantos.
Details on study design:
SCORING SYSTEM: In vitro irritancy score:
The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).
Additionally the opacity and permeability values were evaluated independently to determine whether the test substance induced irritation through only one of the two endpoints. A test substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant.

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Irritant / corrosive response data:
The individual in vitro irritancy scores for the negative controls ranged from -0.2 to 0.1.

The individual positive control in vitro irritancy scores ranged from 100 to 132. All three corneas treated with the positive control were turbid after the 240 minutes of treatment and on one cornea a spot was observed.

The corneas treated with TRANTOS showed opacity values ranging from -2 to 0 and permeability values ranging from 0.021 to 0.034. The corneas were clear after the 240 minutes of treatment with TRANTOS. No pH effect of the test substance was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -1.7 to 0.3 after 240 minutes of treatment with TRANTOS.

Any other information on results incl. tables

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 115 and within the historical positive control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
TRANTOS did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of -0.6 after 240 minutes of treatment.
Finally, it is concluded that this test is valid and that TRANTOS is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.