Registration Dossier

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: Eye irritation was determined according to OECD Guideline 437 (Opacity and permeability of the bovine cornea for the identification of corrosive and highly irritating to the eye).

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Gudeline 437
Principles of method if other than guideline:
The Bovine Corneal Opacity and Permeability test method (BCOP) is an in vitro test method that can be used to classify substances as ocular corrosives and severe irritants. The BCOP uses isolated corneas from the eyes of cattle slaughtered for commercial purposes. Each treatment group (test substance, negative/positive controls) consists of a minimum of three eyes where the cornea has been excised and mounted to a holder. Toxic effects to the cornea are measured as opacity and permeability, which when combined gives an In Vitro Irritancy Score (IVIS) for each treatment group. A substance that induces an IVIS superior or equal to 55.1 is defined as a corrosive or severe irritant.
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent

Test animals / tissue source

Species:
other: Bovine eyes
Strain:
other: Not applicable
Details on test animals or tissues and environmental conditions:
Bovine eyes were collected from a local abattoir and delivered to the laboratory within 4 hours of animal sacrifice and were kept moist and transported chilled to the laboratory.
Preparation of corneas:
Each eyes was dissected into anterior and posterior parts along the coronal meridian. The front part was placed, cornea side down, into a petri dish pre-filled with Phosphate Buffered Saline. Each cornea was mounted in a holder with the endothelial side towards the O-ring of the posterior part of the holder. The compartments were filled with MEM and the corneas were incubated for 60 +-10 minutes in waterbath at 32+-2°C.
Using a blunt needle attached to a 5mL syringe, the media was completely removed from the anterior chamber.
Carrying out the test.

Test system

Vehicle:
water
Controls:
yes
Amount / concentration applied:
A 20% solution of the test substance in water was evaluated for its ocular irritancy potential using corneas
Duration of treatment / exposure:
Test substance (750 μL) is introduced into the anterior chamber through the dosing holes on the top surface of the chamber ( to cover the epithelial side of the cornea), and the holes are subsequently sealed with the chamber plugs during the exposure.
Incubation was carried out at 32+-2°C in a waterbath for 4+-0.25 hours.
Observation period (in vivo):
After incubation, the medium and the test substance was removed from the anterior segment. The epithelium was washed 3 times with MEM containing phenol red. A final wash was performed with MEM wothout phenol red, and the chamber was refilled with MEM.
Opacity measurement and permeability were performed.
Number of animals or in vitro replicates:
3 corneas for test substance, 2 for positive control and 3 for negative control.

Results and discussion

In vivo

Results
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 4 hours
Score:
ca. 173
Reversibility:
not specified
Irritant / corrosive response data:
All three corneas treated with the test article showed a marked increase in corneal opacity. Visual inspection showed an obvious opacity and roughnessof the corneal surface. Two of the three cornea showed minimal change in permeability, while the third cornea presented virtually no barrier to permeability. Based on the measurements obtained, the in vitro score for the test article was 173.

Applicant's summary and conclusion

Interpretation of results:
Category 2A (irritating to eyes) based on GHS criteria
Remarks:
Migrated information
Conclusions:
The test substance is classified as very severe ocular irritant.