Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 to 31 July 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to current OECD test guidelines and GLP principles.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
E-BK105
IUPAC Name:
E-BK105
Details on test material:
- Name of test material (as cited in study report): E-BK105
- Physical state: Black powder, solid
- Lot/batch No.: MB-1
- Expiration date of the lot/batch: July 6, 2015
- Storage condition of test material: Room temperature, in a dark place

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kitayama Labes Co., Ltd.
- Age at study initiation: 10 weeks
- Weight at study initiation: 1869 to 2064 g
- Housing: Aluminium cages (W318xH358xD498) with stainless steel cascade racks, one animal per cage.
- Diet (ad libitum): Pellet diet (LRC4, Oriental Yeast Co., Ltd.), each lot was analysed: contaminants were confirmed to be within acceptable limits of the testing facility
- Water (ad libitum): Well water admixed with sodium hypochlorite (about 0.2 ppm). Analysed every 6 months: contaminants were confirmed to be within acceptable limits of Waterworks law of Japan.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24-26.1
- Humidity (%): 50.6-73.2
- Air changes (per hr): 10 to 20
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated eye
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
Duration of treatment / exposure:
Once dosed, the treated eye was left unwashed in three animals, while in three other animals about 30 seconds after administration the treated eye was washed.
Observation period (in vivo):
1, 24, 48, 72 and 96 hours after administration
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): in 3 of 6 animals, the eye was washed with about 20 ml/eye of distilled water for 30 seconds at a flow rate not damaging to the eye.
- Time after start of exposure: 30 sec.

SCORING SYSTEM: Draize evaluation criteria (acc to OECD 405), Kay & Calandra's ocular irritation evaluation criteria.

TOOL USED TO ASSESS SCORE: hand-slit lamp / fluorescein: at observations 24 hours after administration, the treated eye was observed first by a slit lamp. One drop of fluorescein sodium solution prepared with Japanese pharmacopoeia water for injection to 2 w/v% was then instilled into the cornea and 2-3 seconds later the cornea was washed with about 20 ml of distilled water. The presence or absence of any injury in the cornea was examined again by a slit lamp.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: #1, #2, #3, #4, #5, #6
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Remarks on result:
other: Score in unwashed and washed eyes.
Irritation parameter:
iris score
Basis:
animal: #1, #2, #3, #4, #5, #6
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
2
Remarks on result:
other: Score in unwashed and washed eyes.
Irritation parameter:
conjunctivae score
Remarks:
redness, chemosis and discharge
Basis:
animal: #1, #2, #3, #4, #5, #6
Time point:
other: 24 , 48 and 72 hours
Score:
0
Max. score:
3
Remarks on result:
other: Score in unwashed and washed eyes. Max. score with chemosis is 4.
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal: eye unwashed animals
Time point:
other: 1 hr
Score:
4.7
Max. score:
110
Reversibility:
fully reversible within: 24 hours
Remarks on result:
other: In eye washed animals the score was 4.0 indicating no effect of washing.
Irritant / corrosive response data:
Conjunctival redness and chemosis of score 1 were observed in all animals and discharge of score 1 was observed in one animals of eye unwashed animals at 1 hour after administration. These changes disappeared by 24 hours after administration in all animals. In eye washed animals, conjunctival redness of score 1 was observed in all animals, chemosis of score 1 was observed in two animals, and discharge of score 1 was observed in one animal at 1 hour after administration. These changes disappeared by 24 hours after administration in all animals. Maximum mean total score (MMTS according to Kay & Calandra)) for unwashed and washed eyes was 8 and 7.3, respectively. 24 hours after administration and onward, pigmentation of conjunctiva and partial cornea considered to be test substance related was noted in all animals. The pigmentation in the partial cornea disappeared by 96 hours after administration.
Other effects:
No clinical signs or abnormal body weight gain was observed in any animal throughout the observation period. Observation of the cornea using fluorescein sodium solution revealed no stain spot in any animal.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
E-BK105 had a minimal irritation potential to the rabbit eye under the conditions of this study (OECD 405): irritation was observed only after 1 hr and fully reversible within 24 hours.
Executive summary:

The acute eye irritation or corrosion potential of E-BK105 was assessed in six (3 eye unwashed and 3 eye washed animals) male New Zealand White rabbits (3 eye unwashed and 3 eye washed animals). The test substance was instilled into the right eye of

the animal at 0.1 g. As a result, conjunctival redness and chemosis of score 1 were observed in all animals and discharge of score 1 was observed in one animal of an eye unwashed animal at 1 hour after administration. These changes were fully reversible within 24 hours. In the eye washed animals conjunctival redness of score 1 was observed in all animals, chemosis of score 1 was observed in two animals, and discharge of score 1 was observed in one animal at 1 hour after administration. These effects were also fully reversible within 24 hours. The MMTS of the eye unwashed and washed animals was 4.7 and 4.0, respectively, showing no effect of the eye washing. In conclusion, E-BK105 had minimal eye irritation potential to the rabbit eye under the conditions of this study.