Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 - 15 July 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: OECD guideline 437 “Bovine corneal opacity and permeability (BCOP) test method for identifying ocular corrosives and severe irritants”
Deviations:
no
Qualifier:
according to
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 124. Bovine Opacity and Permeability Assay - SOP of Microbiological Associates Ltd., 1999.
Deviations:
no
Qualifier:
according to
Guideline:
other: Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.
Deviations:
no
Principles of method if other than guideline:
pH: 7.5 (50g/ L)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- test material: BiOI-BiOBr
- Substance type: inorganic
- Physical state: orange powder
- Stability under test conditions: stable
- Storage condition of test material: at room temperature in the dark
- pH: 7.5 (50 g/L)
- Stability at higher temperatures: stable (maximum temperature: 280°C, maximum duration 4 hours)
Specific details on test material used for the study:
pH: 7.5 (50g/ L)

Test system

Vehicle:
physiological saline
Amount / concentration applied:
TEST MATERIAL
- Amount applied (volume or weight with unit): approx. 750 ul of 20% (w/w) test substance solution per cornea

NEGATIVE CONTROL
- Amount applied: 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount applied: 750 µl per cornea
Concentration (if solution): 20% (w/v) Imidazole


Duration of treatment / exposure:
240 minutes
Details on study design:
TEST SITE
- Isolated bovine cornea

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 240 minutes

CONTROLS
Negative control: A negative control, physiological saline (Merck, Darmstadt, Germany) was included to detect non-specific changes in the test system and to provide a baseline for the assay endpoints.
Positive control: 20% (w/w) Imidazole (Merck Schuchardt DHG, Germany) [CAS Number 288-32-4] solution prepared in physiological saline.

SCORING SYSTEM:
- After exposure the cornea is thoroughly rinsed to remove the test substance followed by immediate opacity measurement and permeability evaluation of the cornea. The permeability is evaluated by sodium fluorescein.

- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Remarks:
240 minutes
Run / experiment:
mean of 3 replica
Value:
ca. 0.7
Negative controls validity:
valid
Remarks:
0
Positive controls validity:
valid
Remarks:
79
Other effects / acceptance of results:
The corneas treated showed opacity values ranging from 0 to 1 and permeability values ranging from 0.020 to 0.057. The corneas were clear after the 240 minutes of treatment. No pH effect of the test substance was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from 0.3 to 1.9 after 240 minutes of treatment. The individual in vitro irritancy scores for the negative controls ranged from -0.9 to 0.0. The individual positive control in vitro irritancy scores ranged from 111 to 139. The corneas treated with the positive control were turbid after the 240 minutes of treatment.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
An in vitro eye irritancy test was performed with bismuth oxy-iodide bromide (20% (w/w) solution) using the Bovine Corneal Opacity and Permeability test (BCOP test) according to OECD/ EC guidelines and GLP principles. It was concluded that bismuth oxy-iodide bromide is not irritant in this test.

Executive summary:

An in vitro eye irritancy test was performed with bismuth oxy-iodide bromide (20% (w/w) solution) using the Bovine Corneal Opacity and Permeability test (BCOP test) according to OECD/EC guidelines and GLP principles.

Reliable positive and negative controls were included. The corneas were clear after the 240 minutes of treatment. The in vitro irritancy scores ranged from 0.3 to 1.9 after 240 minutes of treatment. Based on these data, it is concluded that this test is valid and that bismuth oxy-iodide bromide (tested as a 20% (w/w) solution) is not severe irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.