Registration Dossier

Administrative data

Endpoint:
eye irritation
Remarks:
other: in vitro
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Read-across from GLP-compliant guideline study performed with similar substance.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Guideline 437 of July 2013: Bovine Corneal Opacity and Permeability test method for identifying i) chemicals inducing serious eye damage and ii) chemicals not requiring classification for eye irritation or serious eye damage.
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Species:
other: in vitro test
Strain:
other: bovine eyes

Test system

Vehicle:
physiological saline
Amount / concentration applied:
750 µl of test substance, 20% in physiol. saline, positive control solution (20% w/w imidazole in physiol. saline) or negative control solution (physiol. saline) was introduced into the anterior part of each cornea holder. Following application, turning of the holder into horizontal position and its slight rotation ensured uniform distribution of the test substance over the surface of the cornea.
Duration of treatment / exposure:
4 hours (± 5 min.).
Each cornea holder was incubated in a horizontal position at 32°C ± 1°C in a waterbath. Following incubation, the test substance, positive and negative controls were removed and the epithelial surface of the cornea washed, at least three times or until the wash medium (EMEM* with phenol red) was clear and there was no discolouration.

* EMEM, Eagles Minimal Essential Medium
Details on study design:
Each treatment group (test substance, negative/positive controls) comprised 3 corneas from bovine eyes. Each cornea surrounded by ca. 2 to 3 mm of sclera was mounted to a holder. Eyes were maintained and transported to the laboratory and isolated corneas stored in sufficient HBSS* containing 1% penicilin/streptomycin solution until being used. The eyes were used within 4 hours of slaughter. Adequate exposure of the epithelial surface of the cornea to the test substance was provided. Opacity of the cornea was determined by measurement of light transmission through the centre of each mounted cornea using a calibrated opacitometer. Permeability of the cornea was determined by spectophotometric measurement at 490 nm (OD490) of medium from the posterior compartment of the cornea holder after addition of 1 ml of sodium fluorescein solution (4 mg/ml) to the anterior compartment, thus determining the degree of penetration of sodium fluorescein through all corneal cell layers. Prior to the spectophotometric measurement, incubation with fluorescein solution had happened in horizontal position of the cornea holder at 32°C ± 1°C for 90 ± 5 minutes in a waterbath. Then the medium in the posterior compartment was mixed by drawing approximately 2.5 ml gently up and down a 5 ml syringe, with a needle attached, three times. Any solution giving an OD490 value above 1.8 was diluted 1 in 5 with cMEM.

Effects to the cornea (corresponding to possible eye damage) were measured as opacity and permeability, which when combined gave an In Vitro Irritancy Score (IVIS) for each treatment group. A substance that induces an IVIS greater than 55 is defined as a corrosive or severe irritant.

*HBSS Hanks Balanced Salt Solution containing Ca++ and Mg++ without phenol red

Results and discussion

In vivo

Results
Irritation parameter:
other: in vitro irritancy score (IVIS)
Basis:
mean
Remarks:
calculated from the IVIS of each of 3 isolated bovine corneas
Score:
0.9
Remarks on result:
other: IVIS = Corrected Opacity Value + (15 x Corrected permeability OD490 Value)

Any other information on results incl. tables

As outlined in the „Validity Assessment Report“ for the read-across approach (see IUCLID Section 13) read-across from testing data obtained with the UVCB substance WS405777 is considered appropriate for the safety evaluation as well as classification and labelling of the mono-constituent substance WS406663 based on the close chemical similarity between the two substances.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Executive summary:

As outlined in the „Validity Assessment Report“ for the read-across approach (see IUCLID Section 13) read-across from testing data obtained with the UVCB substance WS405777 is considered appropriate for the safety evaluation as well as classification and labelling of the mono-constituent substance WS406663 based on the close chemical similarity between the two substances.

WS406663 does not necessitate any classification regarding eye irritation.