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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:
12.09. – 13. 09. 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
Name of test material (as cited in study report): Solvent Yellow 124
Substance type: organic
Physical state: liquid
Appearance: dark yellow viscous liquid

Composition of test material, percentage of components:
- Analytical purity: 90.0 % (w/w)
- Impurities (identity and concentrations):
4´-[2-((hydroxy)ethyl)ethylamino]azobenzene 3.0 % (w/w)
1,1-bis(N-ethyl[4-(phenylazo)phenyl]aminoethan-2-oxy)ethan 2.5 % (w/w)
- Unknown impurities 4.5 % (w/w)

Lot/batch No.: S2408
Expiration date of the lot/batch: Unlisted
Stability under test conditions: stable
Storage condition of test material: During the study the test substance was stored in glass bottle at laboratory temperature.

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
Open-chamber method was used, because the test substance was viscous liquid. The test substance (enough test substance to completely cover the cornea) was applied directly to the epithelial surface of the cornea using the micropipette. After dosing, the glass window was replaced on the anterior chamber to recreate a closed system.
Duration of treatment / exposure:
10 mins
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done):
The test substance, was removed from the anterior chamber with olive oil (the removal of test substance with water is not possible) and subsequently with EMEM (containing phenol red - the effectiveness of rinsing acidic or alkaline materials). The corneas were given a final rinse with EMEM (without phenol red). The EMEM (without phenol red) was used as a final rinse to ensure removal of the phenol red from the anterior chamber prior to the opacity measurement. The anterior chamber was then refilled with fresh EMEM without phenol red.

- Time after start of exposure:
Procedure scheme:
Selection of corneas, mounting in holders → incubation with EMEM 1hour (32 ± 1°C) → removed EMEM, measurement of baseline opacity → treatment by positive and negative control substance and test substance (incubation 10 min.) → washing epithelium, incubation 2 hour (32 ± 1°C), measurement of opacity after application → application of sodium fluorescein (4 mg/ml), incubation 1.5 hour (32 ± 1°C) → measurement of optical density (490 nm).

TOOL USED TO ASSESS SCORE: fluorescein

SCORING SYSTEM:
Opacity: Opacity values of treated corneas were corrected by subtracting individual background opacity values.

Permeability: Mean OD value of treated corneas was corrected by subtracting the mean OD value of negative control.

IVIS calculation: Resulting mean opacity and OD490 values for each treatment group was combined in an empirically-derived formula to calculate an in vitro irritancy score (IVIS) for each treatment group as follows:
IVIS = mean opacity value + (15 x mean permeability OD490 value)

Decision criteria:
A substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Value:
>= 1.9
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Any other information on results incl. tables

The In Vitro Irritancy Score (IVIS) was computed according the following formula:

IVIS = mean opacity value + (15 x mean permeability OD490 value)

For calculation of IVIS value was used olive oil as negative control because the test substance was removed with olive oil.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The In Vitro Irritancy Score (IVIS) for Solvent Yellow 124 was 1.90. This is less than the limit value of IVIS (55.1), which means that the test substance was not identified as a corrosive or severe irritant.
Executive summary:

The test substance, Solvent Yellow 124, was tested for the evaluation the potential ocular corrosivity or severe irritancy as measured by its ability to induce opacity and increased permeability in an isolated bovine cornea.

The test was performed according to the Method B.47 Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants, Council Regulation (EC) No.1152/2010, published in O.J. L 324, 2010.

The test was performed using nine isolated bovine corneas. The testing was performed on three groups of corneas: test substance treatment group, positive control group and negative control group. Three corneas per group were used.

Open-chamber method was used, because the test substance was viscous liquid. The opacity and permeability of each cornea were measured. The In Vitro Irritancy Score (IVIS) was calculated from the values of opacity and permeability.

The In Vitro Irritancy Score (IVIS) for Solvent Yellow 124 was 1.90. This is less than the limit value of IVIS (55.1), which means that the test substance was not identified as a corrosive or severe irritant.