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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:
2015-02-11 to 2015-02-11
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
no
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
Type:
Constituent
Test material form:
other: liquid

Test animals / tissue source

Species:
other: Bovine
Strain:
not specified

Test system

Vehicle:
unchanged (no vehicle)
Remarks:
(RPMI medium used prior to treatment with test substance)
Controls:
other: 3 corneas treated with saline as negative control and 3 corneas treated with ethanol as positive control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 μl

CONTROLS
- Positive control substance: 100% ethanol
- Negative control substance: physiological saline 0.9% NaCl
Duration of treatment / exposure:
10 minutes incubation at 32 ± 1°C
Observation period (in vivo):
Opacity measurement was performed 2 hours after washing with MEM containing phenol red then rinsing with RPMI; optical density at 490 nm measured after 90 minutes incubation in 4 mg/ml fluoroscein.
Number of animals or in vitro replicates:
3 corneas treated with test substance.
Details on study design:
PREPARATION OF CORNEAS: defective corneas were discarded. Corneas were excised and mounted in corneal holders. Chambers of corneal holder filled with RPMI (without phenol red). The corneas were incubated for one hour at 32 ± 1 °C in a water bath.

The opacitometer was calibrated using empty corneal holders, and linearity was checked.

TREATMENT AND MEASUREMENTS: Initial opacity reading was recorded, and only those with a reading of 7 or below were used. The medium in the anterior chamber was replaced with test item or control. 750 μl of test or control substance was introduced into the anterior chamber and incubated for 10 minutes at 32°C followed by washing with MEM containing phenol red and then rinsed with complete RPMI (without phenol red). The anterior chamber was refilled with complete RPMI and an opacity measurement made after 2 h incubation at 32°C.

After the opacity measurement was performed, the medium was removed from both chambers of the holder. The posterior chamber was refilled with fresh complete RPMI. 1 mL of a 4 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1 °C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer.

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Value:
1.33
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
other: permeability
Run / experiment:
Mean optical density at 490 nm relative to control
Value:
1.33
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
in vitro irritation score
Run / experiment:
opacity and permeability
Value:
1.33
Vehicle controls validity:
valid
Negative controls validity:
valid
Remarks:
0.9
Positive controls validity:
valid
Remarks:
88.29

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The eye irritancy potential of tris(isopropenyloxy)(vinyl)silane has been investigated in a valid in vitro bovine corneal opacity and permeability assay conducted in accordance with OECD 437 and in compliance with GLP. The following mean in vitro irritation score was calculated: 1.33. The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid. It is concluded that the test substance is not irritating to the eye.