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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:
August 31, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Deviations:
not specified
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Source of Bovine Eyes
Eyes from adult cattle (typically 12 to 60 months old) were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter, and were placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 μg/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were prepared immediately on arrival.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
0.75 mL
Duration of treatment / exposure:
10 minutes
Duration of post- treatment incubation (in vitro):
32 ± 1 ºC for 120 minutes

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
Mean
Value:
ca. 29.8
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: No prediction can be made
Other effects / acceptance of results:
The positive control In Vitro Irritancy Score was within the range of 31.6 to 58.7. The positive control acceptance criterion was therefore satisfied. The negative control gave opacity of ≤3.0 and permeability ≤0.077. The negative control acceptance criteria were therefore satisfied.

Any other information on results incl. tables

Results

Corneal Opacity and Permeability Measurement

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in below table:

Treatment

Cornea number

Opacity

Permeability (Optical density)

In Vitro Irritancy Score

Pre-treatment

Post-treatment

Post incubation

Post-incubation - Pre‑treatment

Corrected value

 

Corrected value

Negative control#

1

3

3

4

1

 

0.007

 

 

2

3

2

2

0

 

0.011

 

 

3

4

2

2

0

 

0.007

 

 

 

 

 

 

0.3*

 

0.008¨

 

0.5

Positive control#

4

6

40

37

31

30.7

0.794

0.786

 

5

3

31

33

30

29.7

1.960

1.952

 

6

2

24

26

24

23.7

1.885

1.877

 

 

 

 

 

 

28.0·

 

1.538·

51.1

Test substance

13

1

6

21

20

19.7

0.747

0.739

 

14

3

6

22

19

18.7

0.824

0.816

 

15

1

6

21

20

19.7

0.539

0.531

 

 

 

 

 

 

19.3·

 

0.695·

29.8

* = Mean of the post-incubation-pre‑treatment values                           

# = Control data shared with Envigo - Shardlow study number PS75FR

Corneal Epithelium Condition

The condition of each cornea is given in below table:

Treatment

Cornea Number

Observation

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 Substance

13

Clear

Cloudy

14

Clear

Cloudy

15

Clear

Cloudy

# = Control data shared with Envigo - Shardlow study number PS75FR

The corneas treated with the test substance were clear post treatment and cloudy post incubation. The corneas treated with the negative control substance were clear post treatment and post incubation. The corneas treated with the positive control substance were cloudy post treatment and post incubation.

 

In Vitro Irritancy Score

The In Vitro irritancy scores are summarized as follows:

Treatment

In Vitro Irritancy Score

Test Substance

29.8

Negative Control

0.5

Positive Control

51.1

 

Criteria for an Acceptable Test

The positive control In Vitro Irritancy Score was within the range of 31.6 to 58.7. The positive control acceptance criterion was therefore satisfied. The negative control gave opacity of ≤3.0 and permeability ≤0.077. The negative control acceptance criteria were therefore satisfied.

 

Conclusion

Based on the study results, the study author concluded that no prediction of eye irritation could be made.

Applicant's summary and conclusion

Interpretation of results:
other: No prediction can be made based on EU CLP criteria
Conclusions:
Under study conditions, no prediction could be made about eye irritation potential of the test substance.
Executive summary:

An in vitro study was conducted to determine the eye irritation potential of the the test substance, 'C18-unsatd and C22-unsatd. AAP EDM-ES' (active: 104%), using Bovine Corneal Opacity and Permeability (BCOP) method, according to the OECD Guideline 437 and EU B.47 Method, in compliance with GLP. The undiluted test substance and reference substances were applied to the test system for 10 minutes followed by an incubation period of 120 minutes. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability), were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). The IVIS of the test substance was determined to be 29.8 which is well below the corrosive limit of 55 and above the non-corrosive limit of 3; therefore no predictions could be made. Therefore based on EU CLP criteria, no prediction could be made about eye irrtation potential of the test substance. The IVIS of the positive control was within the range of 31.6 to 58.7, therefore acceptance criterion was satisfied. The negative control gave opacity of ≤3.0 and permeability ≤0.077, therefore the negative control acceptance criteria were satisfied. The quality criteria required for acceptance of results in the test were satisfied. Under the study conditions, no prediction could be made about eye irritation potential of the test substance (Envigo, 2017).