L 131

Eye irritation

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 guideline

Test guideline 1

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

Test guideline 2

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

Test guideline 3

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

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006

Deviations:

no

Test guideline 5

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

Test material information

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.