Registration Dossier

Administrative data

Description of key information

Based on the results of the performed acute dermal toxicity study and the performed BCOP test it is concluded that the substance is not irritating/corrosive to skin and not irritating/corrosive to eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
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.
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
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
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

- 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.

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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

As the acute dermal toxicity study did not indicate skin irritation up to the limit dose level (2000 mg/kg bw), the studies in vitro study for skin corrosion, in vitro study for skin irritation and in vivo study for skin irritation are not performed and is it concluded that the substance is not irritating/corrosive to the skin.

Eye irritation

Using the Bovine Corneal Opacity and Permeability test (BCOP test) the substance 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.


Justification for selection of eye irritation endpoint:
One study available.

Justification for classification or non-classification

Based on the results of the performed acute dermal toxicity study the substance does not need to be classified for skin irritation, in accordance with the CLP Regulation, as no erythema nor oedema was observed during the observation period.

Based on the results of the performed BCOP test the substance does not need to be classified for Eye irritation, in accordance with the CLP Regulation, since the substance induced an IVIS ≤ 3.