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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
12 November 2012 to 22 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant, guideline study, available as an unpublished report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Solid
Details on test material:
- Physical state: Pale Yellow Solid
- Purity: Not applicable - UVCB
- Substance identity: Aluminum, benzoate C16-18-fatty acids complexes
- Batch number: 11074091 + Benzoic acid
- Carbon Content: 65.1%
- Analysis code: A118
- Date recieved: 07 June 2012
- Expiration date: 01 July 2013
- Storage of test material: Room temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Two New Zealand White rabbits were supplied by Harlan Laboratories UK Limited, Hillcrest, Belton, Loughborough, UK. At the start of the study the animals weighed 2.22 or 2.83 kg and were twelve to twenty weeks old. After an acclimatisation period of at least five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23°C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
A volume of 0.1 ml of the test material, which was found to weigh approximately 62 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye.
Duration of treatment / exposure:
Up to 72 hours
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
2
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: Not applicable

The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of cornea involved). For each tissue the score was calculated as follows:
Score for conjunctivae = (A + B + C) x 2
Score for iris = D x 5
Score for cornea = (E x F) x 5
Using the numerical data obtained a modified version of the system described by Kay J H and Calandra J C (1962), J. Soc. Cosmet. Chem. 13, 281 289 (see Appendix 3) was used to classify the ocular irritancy potential of the test material. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test material.
If evidence of irreversible ocular damage is noted, the test material will be classified as corrosive to the eye.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Highest score observed at 1 and 24 hour timepoints
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 24 hours
Remarks on result:
other: Highest score observed at 1 hour timepoint
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Highest score observed at 1 and 24 hour timepoints
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Highest score observed at 1 hour timepoint
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effect noted
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effect noted
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: mean at 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: No effect noted
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: No effect noted
Irritation parameter:
other: Conjunctivae Discharge
Basis:
animal #1
Remarks:
72655 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
other: No effects noted
Irritation parameter:
other: Conjunctivae Discharge
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 1 hour
Remarks on result:
other: Highest score observed at 1 hour timepoint
Irritant / corrosive response data:
Individual and group mean scores for ocular irritation are given in Table 1 and Table 2 in any other information on results section. Individual mean scores for cornea, iris and conjunctivae are given in Table 3.
No corneal or iridial effects were noted during the study.
Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye one hour after treatment with minimal conjunctival irritation noted in both treated eyes at the 24 Hour observation. Both treated eyes appeared normal at the 48 Hour observation.
Other effects:
Both animals showed expected gain in bodyweight during the study (Table 4).

Any other information on results incl. tables

Table 1              Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

72665 Male

72703 Male

IPR= 2

IPR = 1

Time After Treatment

1
Hour

24
Hours

48
Hours

72
Hours

1
Hour

24
Hours

48
Hours

72
Hours

CORNEA

 

 

 

 

 

 

 

 

E = Degree of Opacity

0

0

0

0

0

0

0

0

F = Area of Cornea Involved

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

IRIS

 

 

 

 

 

 

 

 

D

0

0

0

0

0

0

0

0

Score (D x 5)

0

0

0

0

0

0

0

0

CONJUNCTIVAE

 

 

 

 

 

 

 

 

A = Redness

1

1

0

0

2

1

0

0

B = Chemosis

1

1

0

0

2

1

0

0

C = Discharge

0

0

0

0

1

0

0

0

Score (A + B + C) x 2

4

4

0

0

10

4

0

0

Total Score

4

4

0

0

10

4

0

0

IPR=  Initial pain reaction

Interpretation According to the Globally Harmonized System of Classification and Labelling of Chemical

Irreversible eye effects categories

An eye irritant Category 1 (irreversible effects on the eye)is a test material that produces:

-

at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days; and/or

-

at least in 2 of 3 tested animals a positive response of:

corneal opacity³ 3 and/or

iritis³ 1.5r

calculated as the mean scores following grading at 24, 48 and 72 hours after instillation of the test material

Reversible eye effects categories

An eye irritant Category 2A (irritating to eyes)is a test material that produces:

-

at least in 2 of 3 tested animals a positive response of:

corneal opacity³ 1 and/or

iritis³ 1 and/or

conjunctival redness³ 2 and/or

conjunctival oedema (chemosis)³ 2

-

Calculated as the mean scores following grading at 24, 48 and 72-Hour observations after instillation of the test material, and

at

-

which fully reverses within an observation period of normally 21 days

Within this category an eye irritant isconsidered mildlyirritating to eyes (Category 2B)when the effects listed above are fully reversible within 7 days of observation.

Interpretation According to Regulation (EC) No 1272/2008, Relating to the Classification, Labelling and Packaging of Dangerous Substances

        

Category for irreversible eye effects

Category

Criteria

Irreversible effects on the eye

(Category 1)

If, when applied to the eye of an animal, a substance produces:

-  at least in one animal effects on the cornea, iris or conjunctiva that   are not expected to reverse or have not fully reversedwithin an observation periodof normally 21 days;

and/or

-     at least in 2 of 3 animals, a positive response of:

-     cornea opacity³3 and/or

-     iritis>1.5

calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

Category for reversible eye effects

Category

Criteria

Irritating to eyes

(Category 2)

If, when applied to the eye of an animal, a substance produces:

-     at least in 2 of 3 tested animals, a positive response of:

-   cornea opacity³1 and/or

-   iritis>1, and/or

-   conjunctival redness³2 and/or

-   conjunctival oedema (chemosis)³2

-   calculated as the mean scores following grading at 24, 48 and         72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

For those substances where there is pronounced variability among animal responses, this information shall be taken into account in determining the classification.

Table 2              Individual Total Scores and Group MeanScores forOcular Irritation

Rabbit Number

and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

72665 Male

4

4

0

0

72703Male

10

4

0

0

Group Total

14

8

0

0

Group Mean Score

7.0

4.0

0.0

0.0

Table 3            Individual and Mean Scores for Cornea, Iris and Conjunctivae

Rabbit Number and Sex

Time After Treatment

Corneal Opacity

Iridial Inflammation

Conjunctival Redness

Conjunctival Chemosis

72665 Male

24 Hours

0

0

1

1

48 Hours

0

0

0

0

72 Hours

0

0

0

0

Total

0

0

1

1

Mean

0.0

0.0

0.3

0.3

72703 Male

24 Hours

0

0

1

1

48 Hours

0

0

0

0

72 Hours

0

0

0

0

Total

0

0

1

1

Mean

0.0

0.0

0.3

0.3

 

Table 4              Individual Bodyweights and Bodyweight Changes

Rabbit Number
and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

72665 Male

2.83

2.88

0.05

72703 Male

2.22

2.25

0.03

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other:
Conclusions:
The test material, aluminum, benzoate C16-18-fatty acids complexes, produced a maximum group mean score of 7.0 and was classified as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. Aluminum, benzoate C16-18-fatty acids complexes does not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.
Executive summary:

Introduction.

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The assessment was carried out in a GLP-compliant study following OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002) and Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study (Harlan 2013). The study is considered reliable and relevant for use for this endpoint.

Result.

A single application of the test material to the non-irrigated eye of two rabbits produced moderate conjunctival irritation in one treated eye and minimal conjunctival irritation in the other treated eye one hour after treatment with minimal conjunctival irritation noted in both treated eyes at the 24-hour observation. Both treated eyes appeared normal at the 48-hour observation.

Conclusion.

The test material, aluminum, benzoate C16-18-fatty acids complexes, produced a maximum group mean score of 7.0 and was classified as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. Aluminum, benzoate C16-18-fatty acids complexesdoes not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.