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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:
September 22, 2014 - September 23, 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:
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:
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:
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:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006
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
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): L 130
- Appearance: Faint light-green, powdery solid

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
- Corneas that had an initial opacity reading higher than 7 were not used
- Three corneas were used for each treatment group, at random.

REMOVAL OF TEST SUBSTANCE
- Washing: yes, with MEM+phenol red
- Time after start of exposure: 4 hours (at 32 °C)

SCORING SYSTEM:
- After exposure, the cornea is thoroughly rinsed to remove the test substance followed by immediate opacity measurement
- Following the final opacity measurement, permeability of the cornea to Na-fluorescein (Merck) was evaluated, after another incubation period of 90 minutes at 32 °C. OD490 values of less than 1500 were used in the calculation.
- 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:
0.4
Remarks on result:
other: Negative control: 0; Positive control: 139

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

99

2.673

139

L 130

0.3

0.004

0.4

- Individual in vitro irritancy scores:

 

Eye

In vitro Irritancy Score

Negative control

1

2.1

2

-2.1

3

0.1

Positive control

4

143.4

5

151.5

6

123.4

L 130

7

-0.9

8

1.0

9

1.1

 

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 130 (a solid; tested in a 20% (w/w) suspension) did not induce ocular irritation (mean in vitro irritancy score of 0.4), L 130 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 130 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 of 0.4 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.