Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 6, 2014 - October 7, 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Version / remarks:
(2013)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Version / remarks:
(2010)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: The Ocular Toxicity Working Group (OTWG) of ICCVAM and NICEATM, Background Review Document (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The BCOP Test Method, March 2006
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006
Deviations:
no
Qualifier:
according to guideline
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
GLP compliance:
yes

Test material

Constituent 1
Details on test material:
- Name of test material (as cited in study report): L 131
- Appearance: Faint light-green powder

Test system

Vehicle:
physiological saline
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 750 µL of a 20% (w/w) suspension in the vehicle per cornea

NEGATIVE CONTROL
- Amount applied: 750 µL of the vehicle per cornea

POSITIVE CONTROL
- Amount applied: 750 µL of a 20% (w/v) Imidazole solution in the vehicle per cornea
Duration of treatment / exposure:
4 hours
Details on study design:
TEST SITE
- Isolated bovine cornea
- Three corneas were used for each treatment group

REMOVAL OF TEST SUBSTANCE
- Washing: yes, with MEM with phenol red
- Time after start of exposure: 4 hours

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 mean opacity and mean permeability values (OD490) were used for each treatment group to calculate the mean in vitro score:
Mean in vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).
- Opacity and permeability values were also evaluated individually

TOOL USED TO ASSESS SCORE:
- Opacitymeter and microplate reader

DATA EVALUATION:
- A test substance that induces a mean IVIS >55 is classified with Eye Irr. Cat. 1 in accordance with the CLP Regulation
- A test substance that induces a mean IVIS ≤ 3 is not classified for Eye irritation in accordance with the CLP Regulation
- For a test substance that induces a mean IVIS of >3 - ≤55, it cannot be concluded whether the substance needs to be classified or not and for these substances, more (in vivo) information is needed

Results and discussion

In vivo

Results
Irritation parameter:
other: In vitro irritancy score (IVIS)
Basis:
mean
Time point:
other: 4 hours
Score:
1.3
Remarks on result:
other: Negative control: 0; Positive control: 144

Any other information on results incl. tables

Summary of opacity, permeability and in vitro scores:

Treatment

Mean

Opacity

Mean

Permeability

Mean In vitro Irritation Score

Negative control

0

0.000

0.0

Positive control

126

1.238

144

L 131

1.0

0.018

1.3

Individual in vitro irritancy scores:

Eye

In vitro Irritancy Score

Negative control

1

1.5

2

-1.8

3

0.3

Positive control

4

153.2

5

142.7

6

136.8

L 131

7

1.1

8

3.1

9

-0.4

 

No pH effect of the test substance was observed on the rinsing medium.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the results of a Bovine Corneal Opacity and Permeability test in which L 131 did not induce ocular irritation (mean in vitro irritancy score of 1.3), L 131 does not need to be classified for Eye irritation in accordance with the CLP Regulation.
Executive summary:

Using the Bovine Corneal Opacity and Permeability test (BCOP test) L 131 was screened for its eye irritancy potential in accordance with OECD 437 and according to GLP principles. The substance was applied as a 20% (w/w) suspension (750 µL) directly on top of the corneas. Adequate negative and positive controls were included. The substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score (IVIS) of 1.3 after 240 minutes of treatment. Since the substance induced a mean IVIS ≤ 3, the substance does not need to be classified for Eye irritation in accordance with the CLP Regulation.