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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was performed according to the recommended guidelines and GLP, and without deviations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: The OTWG on the ICCVAM and the NICEATM, BRD: current status of in vitro test methods for identifying ocular corrosives and severe irritants: The BCOP Test Method, March 2006.
Qualifier:
according to guideline
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 124. Bovine Opacity and Permeability assay - SOP of Microbiological Associates Ltd., 1999.
Qualifier:
according to guideline
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.
Principles of method if other than guideline:
In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimise the need of in vivo testing. As a consequence a validated and accepted in vitro test for eye irritation should be performed before in vivo tests are conducted. One of the proposed in vitro eye irritation tests is the Bovine Corneal Opacity and Permeability (BCOP) test. The BCOP assay measures two important components which are predictive of irritation, corneal opacity and permeability. Although the BCOP test method is not yet validated, the EU national regulatory authorities accept positive outcomes (severe eye irritants) of this method for classification and labelling (R41). Where no positive result is obtained, an in vivo test is subsequently required.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Species:
other: isolated corneas from bovine eyes

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
750 µl undiluted substance
Duration of treatment / exposure:
Corneas were incubated in a horizontal position for 10 +/- 1 minutes at 32 +/- 1°C
Observation period (in vivo):
After the incubation the control or test substance was removed and the epithelium was washed at least three times with cMEM. The medium in the posterior compartment was removed and both compartments were refilled with fresh cMEM. Subsequently the corneas were incubated for 120 +/-10 minutes at 32 +/- 1°C. After the completion of the incubation period opacity determination was performed.

Results and discussion

In vivo

Results
Irritation parameter:
other: in vitro irritancy scores: opacity and permeability
Basis:
mean
Time point:
other: 10 minutes
Score:
0
Max. score:
0.2
Reversibility:
not specified

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The mean in vitro irritancy score (uncorrected) obtained with the negative control was less than 3.1 indicating that the negative controls did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (10% w/v Benzalkonium Chloride) was within the historical positive control data range (Appendix I, table 5). It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The in vitro irritancy scores of the corneas treated with the test substance ranged from -0.1 to 0.2 with a mean of 0.0. Since all in vitro irritancy scores were less than 3.1, the test substance was considered non irritant.

Finally, it is concluded that this test is valid and that the test substance is non irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.