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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
26 August 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Followed established Guideline and GLP requirements

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
In vitro test in step wise procedure of Guideline 405
Qualifier:
according to
Guideline:
other: OECD No. 437 (2009) "Bovine Corneal Opacity and Permeability Assay"
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): SD10
- Physical state: Clear, colorless liquid
- Analytical purity: 96.9%
- Lot/batch No.: 9147-192-3a
- Expiration date of the lot/batch: 11 April 2012
- Storage condition of test material: room temperature in dark, under nitrogen

Test animals / tissue source

Species:
other: bovine corneas
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: local abbatoir
- Age at study initiation: adult cattle, eyes obtained from freshly slaughtered animals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): eyes transported in Hanks' Balanced Salt Solution supplemented by Penicillin/Streptomycin and placed on ice packs. Eyes refrigerated on arrival to lab, and used within 24 hours.


Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL test item or control item applied to each cornea.
- Concentration (if solution): Negative control was 0.9% sodium chloride solution.

Duration of treatment / exposure:
After test item or control item applied to cornea in the BCOP holder and the holder tilted to assure uniform applicaiton to the cornea, the holder containing the cornea was incubated at 32 +/- 1 degrees C for 10 minutes. The cornea was then rinsed three times with complete Minimum esssential medium (MEM) containing phenol red before a final rinse with complete MEM.
Observation period (in vivo):
A pre-treatment opacity reading of each cornea was taken before the exposure period. A post-treatment opacity reading was taken and each cornea observed visually after the rinsing with complete MEM. The holders were then incubated at 32 +/- 1 degrees C for 120 minutes +/- 10 minutes. After that incubation, a final opacity reading was taken and each cornea visually observed.Following the final opacity reading, the medium was removed from the anterior chamber and replaced with 1 ml sodium fluorescein (4 mg/ml). The holders were incubated for 90 minutes at 32 +/- 1 degrees C. Medium from the posterior chamber was used for optical density reading (a sample of 360 ul represented each cornea.
Number of animals or in vitro replicates:
Three corneas each for test item, negative control and positive control
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, after 10 minutes, the corneas were rinsed with MEM plus phenol red three times before a final rinse with complete MEM
- Time after start of exposure: 10 minutes of exposure

SCORING SYSTEM: Opacity reading using calibrated opacitometer; permeability measured by optical density at 492 nm;

TOOL USED TO ASSESS SCORE: Calibrated opacitometer; 96 well plate for measuring optical density at 429 nm.

Results and discussion

Results of in vivo study
Irritation parameter:
other: In Vitro Irritation Score (IVIS)
Basis:
other: mean value for treated cornea group
Score:
ca. 66
Remarks on result:
other: The IVIS score was greater then 55, thus the test item is a severe irritant or corrosive
Irritant / corrosive response data:
See tables in results section for individual corneal and mean treatment values for opacity, permeability, corrected values of each, and the resulting IVIS scores.

Any other information on results incl. tables

Individual and Mean Corneal Opacity Measurements

Opacity

Treatment

Cornea Number

Pretreatment/

Postreatment/Post Incubation

Post incubation - Pretreatment

Corrected Value

Negative Control

1

1

1

2

1

2

1

1

1

0

3

2

2

3

1

Mean

0.7

Positive Control

4

2

28

28

26

25.3

5

1

27

27

26

25.3

6

2

25

25

23

22.3

Mean

24.3

Test Item

7

3

5

9

6

5.3

8

1

10

13

12

11.3

9

1

4

18

17

16.3

Mean

11.0

Individual and Mean Permeability Measurements and Resulting IVIS

Permeability (Optical Density, OD)

Treatment

Cornea Number

Corrected Value

In vitro Irritancy Score (IVIS)

Negative Control

1

0.057

2

0.127

3

0.056

Mean

0.080

1.9

Positive Control

4

2.860

2.780

5

2.650

2.570

6

3.350

3.270

Mean

2.873

67.4

Test Item

7

4.545

4.465

8

3.665

3.585

9

3.025

2.945

Mean

3.665

66.0

Corneal Epithelium Post Treatment and Post Incubation

Observation

Treatment

Cornea Number

Post Treatment

Post Incubation

 

Negative Control

1

Clear

Clear

 

2

Clear

Clear

 

3

Clear

Clear

 

Positive Control

4

Cloudy

Cloudy

 

5

Cloudy

Cloudy

 

6

Cloudy

Cloudy

 

Test Item

7

Clear

Cloudy

 

8

Cloudy

Cloudy

 

9

Clear

Cloudy

 

Applicant's summary and conclusion

Interpretation of results:
highly irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: Criteria in OECD No. 437
Conclusions:
Following assessment of all endpoint data in the Bovine Corneal Opacity and Permeability Assay, the test item was considered to have the potential to cause severe ocular irritancy in vivo. The in vivo eye irritation study is therefore not needed.

According to CLP (Reg. 1907/2006 and amendments) the substance is classified as causing irreversible eye effects (category 1) based on this in vitro result.

(Reg. 1907/2006 and amendments) the substance is classified as posing the risk of serious damage to eyes (R41).
Executive summary:

Following assessment of all endpoint data in the Bovine Corneal Opacity and Permeability Assay, the test item was considered to have the potential to cause severe ocular irritancy in vivo. The in vivo eye irritation study is therefore not needed. The In Vitro Irritancy Score, calculated on changes in opacity and permeability (optical density), was 66.0 for the test item compared to 67.4 for the positive control item (ethanol). According to the OECD Guideline No. 437, an IVIS of 55 or greater is interpreted as the test item has the potential for severe ocular irritancy in vivo.

According to CLP (Reg. 1907/2006 and amendments) the substance is classified as causing irreversible eye effects (category 1) based on this in vitro result.

(Reg. 1907/2006 and amendments) the substance is classified as posing the risk of serious damage to eyes (R41).