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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 December 2013 - 17 February 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

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:
solid: particulate/powder

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: breeder: CEGAV, Argenvilliers, France.
- Age at study initiation: 2 to 4 months old on the day of treatment
- Fasting period before study: no
- Housing: individually housed in noryl cages
- Diet: 110 pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: 31 January 2014 to 17 February 2014

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: untreated right eye served as a control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.1 g/animal was used.
Duration of treatment / exposure:
Not applicable: single application followed by rinsing performed just after the 1-hour scoring.
Observation period (in vivo):
1, 24, 48 and 72 h up to the complete reversibility (Day 5).
Number of animals or in vitro replicates:
Three males
Details on study design:
REMOVAL OF TEST SUBSTANCE: Yes

SCORING SYSTEM: Draize scale.

- Conjunctival chemosis (lids and/or nictitating membranes):
0 no swelling
1 any swelling above normal
2 obvious swelling with partial eversion of lids
3 swelling with lids about half-closed
4 swelling with lids more than half-closed

- Conjunctival redness (palpebral and bulbar conjunctivae, excluding the cornea and iris):
0 blood vessels normal
1 blood vessels definitely hyperemic (injected)
2 diffuse, crimson colour, individual vessels not easily discernible
3 diffuse, beefy red

- Iris lesions
0 normal
1 markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia,or injection, any or a combination of any there findings, but, iris still reacting to light (sluggish reaction is positive)
2 no reaction to light, haemorrhage, gross destruction (any or all of these)

- Corneal intensity of opacity (direct examination and, if necessary, with an UV lamp)
0 no ulceration or opacity
1 scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible
2 easily discernible translucent area, details of iris slightly obscured
3 nacreous areas, no details of iris visible, size of pupil barely discernible
4 opaque cornea, iris not discernible through the opacity

- Corneal area of opacity (direct examination and, if necessary, with an UV lamp)
1 one quarter (or less) but not zero
2 greater than one quarter but less than a half
3 greater than one half but less than three quarters
4 greater than three quarters up to whole area

- Any other lesions observed were noted

TOOL USED TO ASSESS SCORE: UV lamp after instillation of 0.5% sodium fluorescein solution

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Remarks:
(intensity)
Basis:
animal: #1, 2 and 3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal: #1, 2 and 3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: day 2
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: day 3
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 24, 48 and 72 h (mean)
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: day 3
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #2
Time point:
other: 24, 48 and 72 h (mean)
Score:
1.3
Max. score:
3
Reversibility:
fully reversible within: day 5
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 24, 48 and 72 h (mean)
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: day 3
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #3
Time point:
other: 24, 48 and 72 h (mean)
Score:
1.3
Max. score:
3
Reversibility:
fully reversible within: day 5
Irritant / corrosive response data:
No ocular reactions were observed in the right untreated control eye.
In the left treated eye, slight chemosis (grade 1) and moderate redness of the conjunctiva (grade 2) were observed in all animals on Day 1. These reactions persisted on Day 2 in 2/3 animals whereas slight redness of the conjunctiva (grade 1) only was noted on Day 2 in the third animal. Then, slight redness of the conjunctiva (grade 1) was still noted until Day 4 on 2/3 animals.
No Iris or corneal lesions were noted in any animals all over the study period.

Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:
. chemosis: 0.0, 0.3 and 0.3; showing slight but no significant eye irritation,
. redness of the conjunctiva: 0.3, 1.3 and 1.3; showing slight but no significant eye irritation,
. iris lesions: 0.0, 0.0 and 0.0; showing no eye irritation,
. corneal opacity: 0.0, 0.0 and 0.0; showing no eye irritation.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
EKKE was slightly irritant when administered by ocular route to rabbits. The findings were all reversible in animals by Day 5.
Accordingly, the test item, should not be classified as irritating to the eyes according to the CLP and GHS criteria.
Executive summary:

The potential irritant properties for the eye of EKKE was evaluated following a single administration to rabbits. This study was conducted in compliance with OECD Guideline No. 405 and the principles of Good Laboratory Practices. EKKE was first administered to a single male New Zealand White rabbit. As mean value from grading at 24, 48 and 72 hours after instillation was < 2 for conjunctival edema (chemosis) and for conjunctival redness and < 1 for iris lesion and for corneal opacity, the test item was administered in the left eye of two other animals. The test item was administered inthe conjunctival sac of the left eye. The right eye remained untreated and served as control. A quantity of 0.1 g/animal was used.

For all animals,a local anaesthetic was used prior to treatment. Just after the 1-hour scoring, both eyes were rinsed with a sterile isotonic saline solution (0.9% NaCl). Each animal was observed at least once a day for mortality and clinical signs. Ocular reactions were observed approximately 1, 24, 48 and 72 hours after the instillation and then daily until the reversibility of the ocular reactions. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. Body weight was recorded on the day of treatment and at the end of the evaluation of ocular reactions. On completion of the observation period, the animals were sacrificed then discarded without macroscopic post-mortem examination.  

No unscheduled deaths occurred during the study and no clinical signs were noted in any animals. The body weight of the animals was unaffected by the test item treatment. No ocular reactions were observed in the right untreated control eye. In the left treated eye, slight chemosis and moderate redness of the conjunctiva were observed in all animals on Day 1. These reactions persisted on Day 2 in 2/3 animals whereas slight redness of the conjunctiva only was noted on Day 2 in the third animal. Then, slight redness of the conjunctiva was still noted until Day 4 on 2/3 animals.

No iris or corneal lesions were noted in any animals all over the study period.

Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:

- chemosis: 0.0, 0.3 and 0.3; showing slight but no significant eye irritation,

- redness of the conjunctiva: 0.3, 1.3 and 1.3; showing slight but no significant eye irritation,

- iris lesions: 0.0, 0.0 and 0.0; showing no eye irritation,

- corneal opacity: 0.0, 0.0 and 0.0; showing no eye irritation.

EKKE was slightly irritant when administered by ocular route to rabbits. The findings were all reversible in animals by Day 5. Accordingly, the test item, should not be classified as irritating to the eyes according to the CLP and GHS criteria.