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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
3 January 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented GLP study conducted according to OECD guideline 437.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: aqueous solution
Details on test material:
- Name of test material (as cited in study report): Zirconium acetate solution
- Substance type: Clear colourless solution
- Physical state: Liquid
- Analytical purity: > 99%
- Composition of test material, percentage of components: aqueous solution containing 40.7% zirconium acetate anhydrous
- Purity test date: 2012-07-30
- Lot/batch No.: 12/201
- Expiration date of the lot/batch: 31 July 2013
- Storage condition of test material: Ambient condition

Test animals / tissue source

Species:
other: Bovine corneas
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Butcher Service s.r.l.- Mattatoio no. 2067 M
- Age at study initiation: 6-12 months of age at collection of eyes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 32 ± 1°C

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
Duration of treatment / exposure:
Corneas were exposed in horizontal position for 10 ± 1 minutes
Observation period (in vivo):
Corneas were observed through chamber glasses and pertinent observation recorded (e.g. tissue peeling/exfoliation, residual test substance, non-uniform opacity pattern etc.). During the post-exposure period, corneas were maintained in horizontal position, for futher 2 hours ± 10 minutes, incubated in a liquid bath at 32 ± 1°C.
Number of animals or in vitro replicates:
Three corneas per treatment (test item, negative and positive control).
Details on study design:
The test item was used in the form supplied, without any further dilution.
Positive control: 1% (w/v) sodium hydroxide in water
Negative control: Physiological saline (0.9% NaCl)

After 10 ± 1 minutes of exposure the corneas were rinsed thoroughly with complete Minimum Essential Medium Eagle's (MEM) EMEM with phenol red. A final wash with prewarmed complete EMEM without phenol red was carried out. After the incubation the opacity of all corneas was determined. On completion of the opacity measurements, EMEM was removed from both chambers (anterior first). The posterior chamber was re-filled with fresh complete EMEM without phenol red maintained at approximately 32°C. The anterior chamber was treated with 1 mL aliquot of 4 mg/mL sodium fluorescein solution in DPBS. The corneas were horizontally incubated in a liquid bath at 32 ± 1 °C for approximately 90 minutes. After the incubation, the medium was removed from the posterior chamber and the optical density of each sample was measured using a verified spectrophotometer at 490 nm. The linear range of a reference curve 0-20 µg/mL is carried out at RTC approximately once a year.
Quality controls of the same fluorescein solutions were prepared in complete EMEM without phenol red at concentrations of 2 and 10 µg/mL and were assayed in order to ensure that the fluorescein calibration curve was still acceptable at the time of the experiment.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: opacity
Basis:
mean
Score:
67.3
Max. score:
79
Remarks on result:
other: Standard Deviation 11.5
Irritation parameter:
other: permeability
Basis:
mean
Score:
0.023
Max. score:
0.037
Remarks on result:
other: Standard Deviation 0.0125
Irritation parameter:
other: in vitro irritancy score
Score:
67.6
Irritant / corrosive response data:
Alterations of cornea opacity were recorded during the study when compared to those of negative control. No significant alterations of cornea permeability were noted.

Any other information on results incl. tables

The negative and positive controls gave the expected results. The test is therefore considered as valid.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the OECD Guideline No. 437, the test material should be classified as corrosive or severely irritant to the eye.