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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2010
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented, according to accepted guidelines for a read-across substance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Guideline 437 Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants (BCOP Test)
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): "Glucose Syrups Wheat Hydrolized", dried wheat glucose syrup, Glucodry 385, dried glucose syrup.
- Physical state: Solid (white, slightly hygroscopic powder).
- Analytical purity: Certificate of Analysis attached in Appendix 4; however, information was illegible.
- Lot/batch No.: 5095187.
- Expiration date of the lot/batch: June 2011.
- Storage condition of test material: At room temperature and no protection from light.

Test animals / tissue source

Species:
other: Bovine cornea
Strain:
not specified
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Slaughterhouse, Klaus Grandits", Ungerbachstr. 10, A-2860 Kirchschlag.
- Age at study initiation: Isolated corneas from the eyes of cows and bulls at the age of 20 months.
- Bovine corneas were equilibrated for 1 hour to achieve normal metabolic activity.

Test system

Vehicle:
water
Remarks:
deionised water (sterile)
Controls:
yes
Amount / concentration applied:
The test substance was administered at a concentration of 20% in deionised water (sterile). Bovine corneas were exposed to 750 µL of test substance.
Duration of treatment / exposure:
The corneas were exposed to the test substance for 4 hours.
Observation period (in vivo):
None.
Number of animals or in vitro replicates:
3 corneas per treatment.
Details on study design:
- Negative control: deionised water, sterile.
- Positive control: 20% imidazole (CAS-No. 288-32-4).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the exposure period, substance was removed from the anterior chamber and the epithelium was washed 3 times with Eagle's Minimum Essential Medium (EMEM)+ to determine the effectiveness of rinsing acidic or alkaline material and to remove substance residues. cEMEM was used as a final rinse to ensure removal of the phenol red from the anterior chamber prior to the opacity measurement.
- Time after start of exposure: 4 hours.

SCORING SYSTEM: Using opacity and permeability data, an in vitro irritancy score (IVIS) was calculated.
IVIS = mean opacity value + (15 x mean permeability OD490 value)

According to the guidelines, an irritation potential of a test substance is predicted if the mean IVIS of 3 individual corneas exposed to the test substance is ≥55.1.

TOOL USED TO ASSESS SCORE: Opacity was measured quantitatively with the aid of an opacitometer. For the purpose of permeability assessment, fluorescein solution was filled into the anterior chambers of the cornea holders followed by an incubation period of 90 minutes. The amount of sodium fluorescein that crossed into the posterior chambers was quantitatively measured with a spectrophotometer at OD490.

Results and discussion

In vivo

Results
Irritation parameter:
other: Not applicable
Irritant / corrosive response data:
Not applicable
Other effects:
Not applicable

Any other information on results incl. tables

Since the IVIS of the negative control was 8.4, instead of 7.4, which is the upper limit of the historical negative control data, the study was repeated.

The IVIS for "Glucose Syrups Wheat Hydrolized" was 1.1. IVIS of the negative control was 3.3 and 105.8 for the positive control, thus demonstrating the validity of the experiment.

Table 1: Opacity, permeability, and IVIS values

Substance

Opacity

Permeability (1x)

Permeability (15x)

IVIS

Individual

Mean

SD

Individual

Mean

SD

Mean

SD

Individual

Mean

SD

Aqua dest (NC)

6.6

3.3

3.5

0.001

0.000

0.007

0.000

0.098

6.6

3.3

3.4

3.5

-0.007

3.3

-0.3

0.006

-0.2

20% Imidazole (PC)

59.5

66.9

6.5

2.991

2.591

0.681

38.867

10.220

104.4

105.8

10.1

69.4

1.804

96.5

71.8

2.978

116.4

Test substance

-0.9

-0.1

0.8

0.008

0.078

0.122

1.175

1.833

-0.8

1.1

2.2

0.5

0.008

0.6

0.2

0.219

3.5

IVIS = in vitro irritancy score; NC = negative control; PC = positive control; SD = standard deviation.

Applicant's summary and conclusion

Interpretation of results:
other: unable to classify
Remarks:
Criteria used for interpretation of results: other: CLP (EC 1272/2008)
Conclusions:
According to the Guidance on the Application of the CLP Criteria (ECHA Reference ECHA-09-G-02-EN), a positive result in the BCOP is classified as serious eye damage Category 1, while negative results in the BCOP must be followed by further testing.