Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23.12.2010-14.04.2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was performed in compliance with the Good Laboratory Practice (GLP) regulations (revised in 1997, ENV/MC/CHEM(98)17). The method followed that described in the OECD Guidelines for Testing of Chemicals, Updated Guideline No 437 Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants (September, 2009).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD 437
Deviations:
no
Principles of method if other than guideline:
none
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Species:
other:
Strain:
other:
Details on test animals or tissues and environmental conditions:
no data

Test system

Vehicle:
physiological saline
Controls:
no
Amount / concentration applied:
20 % (w/v) test substance in saline
Duration of treatment / exposure:
240 min
Observation period (in vivo):
none
Number of animals or in vitro replicates:
none
Details on study design:
according to guideline

Results and discussion

In vivo

Results
Irritation parameter:
other: in vitro irritation score
Basis:
mean
Time point:
other: 240 min
Score:
87.83
Max. score:
112.95
Reversibility:
other: not applicable
Irritant / corrosive response data:
positive control: 254.73
negative control: 0.0
Other effects:
none

Applicant's summary and conclusion

Interpretation of results:
corrosive
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the test conditions, the test item is considered to be corrosive in vitro.
Executive summary:

Purpose

This study should provide a rational basis for risk assessment in humans. Ocular contact is a possible route of human exposure. This in vitro study was performed to assess the corneal irritation and damage potential of the test item by means of the BCOP assay using fresh bovine corneae.

Study Design

Damage is assessed by quantitative measurements of changes in corneal opacity and permeability with an opacitometer and a visible light spectrophotometer, respectively. Both measurements are used to calculate an in vitro irritation score, which is used to assign an in vitro irritancy category. After a first opacity measurement of the fresh bovine corneae (t0), the test item/saline (0.9% (w/v) NaCl in deionised water) stock, the positive, or the negative controls were applied to the epithelial surface of three corneae each and incubated for 240 minutes at 32 +/- 2 °C. After the incubation phase, the cornea were each rinsed and opacity was measured again (t240). Thereafter the permeability of the corneae was determined. Therefore, 1 mL of a fluorescein solution was added to the anterior chamber of the corneal holder and after 90 min incubation in a horizontal position at 32 +/- 2 °C, the permeability of the corneae was measured quantitatively using the increase in fluorescence in the posterior chamber medium at 490 nm.

Results

The negative control (0.9% NaCl solution) yielded no increase of opacity nor permeability of the corneae, while the the positive control (10% (w/v) Benzalconium chloride) showed clear opacity and distinctive permeability of the corneae, thus demonstrating the validity of the assay. The test item caused an increase in opacity of the corneae, but permeability of the corneae was not observed. The calculated mean in vitro score was 87.83. Based on the irritatiion schema, the test item shows severe eye irritant properties in vitro.

Conclusion

The study performed shows that the test item is considered to be a severe eye irritant in vitro under the described conditions.