Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
03 April 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well conducted and well described study in accordance with GLP and OECD Guideline 437 without any deviation.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

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
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Camphor white oil
- Physical state: clear colorless liquid
- Analytical purity: Conforms to Standard
- Lot/batch No.: 160341
- Storage condition of test material: Room temperature in the dark

Test animals / tissue source

Species:
other: Bovine eyes
Details on test animals or tissues and environmental conditions:
Not applicable

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): Undiluted
Duration of treatment / exposure:
- The undiluted test item was applied in BCOP chamber for 10 minutes followed by an incubation period of 120 ± 10 minutes.
Observation period (in vivo):
- The condition of the cornea was visually assessed immediately after rinsing and at the final opacity measurement.
Number of animals or in vitro replicates:
Not applicable
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete minimum essential medium (MEM) containing phenol red before a final rinse with complete MEM. The anterior chamber was refilled with fresh complete MEM.

EVALUATION OF RESULTS:
- Results from the two test method endpoints, opacity and permeability, were combined in an empirically derived formula to generate an in vitro irritancy score.
- The following formula was used to determine the in vitro irritancy score:
In Vitro Irritancy Score = mean opacity value + (15 x mean OD492 value)
Additionally, the opacity and permeability values were evaluated independently to determine whether the test item induced a response through only one of the two endpoints.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: In vitro irritancy score for negative control
Basis:
mean
Remarks:
3 corneas
Score:
1.4
Reversibility:
other: not applicable
Irritation parameter:
other: In vitro irritancy score for test item
Basis:
mean
Remarks:
3 corneas
Score:
3.4
Reversibility:
other: not applicable
Irritation parameter:
other: In vitro irritancy score for positive control
Basis:
mean
Remarks:
3 corneas
Score:
38.7
Reversibility:
other: not applicable
Irritant / corrosive response data:
Corneal epithelium condition: The corneas treated with the test item were clear post treatment and post incubation. The corneas treated with the negative control item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation.
Other effects:
None

Any other information on results incl. tables

See the attached document for tables of results

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the test conditions, Camphor white oil is considered not to be an ocular corrosive or severe irritant. No definitive conclusion can be established since the IVIS was > 3 and ≤ 55 (test item is not predicted as causing serious eye damage and not classified for eye irritation or serious eye damage).
Executive summary:

In an in vitro eye irritation study conducted according to the OECD Guideline 437 and in compliance with GLP, 3 bovine corneas were applied with 0.75 mL of undiluted Camphor white oil for 10 minutes at 32 ±1 ºC followed by an incubation period of 120 ± 10 minutes at 32 ±1 ºC. Negative and positive control items were tested concurrently. After incubation the BCOP holders were removed from the incubator and a final opacity reading was taken. Each cornea was visually observed. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an in vitro irritancy score.

The corneas treated with the test item were clear post treatment and post incubation. The corneas treated with the negative control item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation. The in vitro irritancy score for negative and positive controls were 1.4 and 38.7, respectively and the values were within the respective ranges and therefore the study was considered valid. The in vitro irritancy score for the test item was 3.4, which is considered not to be an ocular corrosive or severe irritant.

 

Under the test conditions, Camphor white oil is considered not to be an ocular corrosive or severe irritant. No definitive conclusion can be established since the IVIS was > 3 and ≤ 55 (test item is not predicted as causing serious eye damage and not classified for eye irritation or serious eye damage).

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.