Dodeca (lithium, sodium)-2-({4-[4-(4-{bis[alkyl-(sulfonatoalkoxy)-4-({-sulfonato-[(substituted-phenyl)diazenyl]phenyl} diazenyl)anilino]-triazin-yl}-6-[alkyl-(sulfonatoalkoxy)-4-(sulfonato--[(substituted-phenyl)diazenyl]phenyl)diazenyl)anilino]piperazin-yl)-triazin-ylamino]-alkyl-5-(sulfonatoalkoxy)phenyl}diazenyl)-5-[(sulfonatophenyl)diazenyl]benzensulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 16 - July 31, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to 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

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): E-BW102
- Physical state: Reddish brown powder
- Storage condition of test material: At room temperature, dark place
- Stability under storage conditions: No data

Test animals / tissue source

Species:

rabbit

Strain:

other: Kbs:NZW (Healthy)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kitayama Labes Co., Ltd.
- Age at study initiation: Animals used within the study were 10 weeks old *
- Weight at study initiation: body weights were 1961 to 2127 g *.
- Housing: Individually housed in labeled aluminium cages with stainless-stell cascade racks (W318 x H358 x D498 mm).
- Diet (e.g. ad libitum): Pelleted diet (LRC4, Oriental Yeast Co., Ltd., lot 100420)
- Water (e.g. ad libitum): Free access to well water.
- Acclimatization period: 6 days from receipt.

* Animal specifications (sex, age and body weight) are specified in the attached table.

Animals without any abnormality in the cornea, iris or conjunctiva were selected.

Results of analysis for diet was provided by Oriental Yeast Co., Ltd., and the contaminants in the diet were confirmed to be within the acceptable limits established by the test facility. The water was analyzed every 6 months at Nichigo Kyushu Co., Ltd. in compliance with the water quality standard (Waterworks Law) of MHLW, Japan. The contaminants in the water were confirmed to be within the acceptable limits.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24.0 -26.1
- Humidity (%): 50.6 - 73.2
The accommodation equipment was washed with a cage washer. The animal room was swept and wiped every day with a mop soaked in disinfectant.
- Air changes (per hr): 10 to 20 times
- Photoperiod (hrs dark / hrs light): 12/12

IN LIFE DATES: From: July 27, 2010 To: July 31, 2010

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: One eye of each animal remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL- Amount applied: 0.1g

Duration of treatment / exposure:

Single instillation on Day 1.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48, 72 and 96 hours after instillation of the test substance.

Number of animals or in vitro replicates:

6 males

Details on study design:

REMOVAL OF TEST SUBSTANCE
-Washing: In the remaining 3 animals, about 30 seconds after administration, the right eye was washed with about 20 mL/eye of distilled water for 30 seconds at a flow rate not damaging the eye.

OBSERVATIONS
- Clinical observation: Observation was performed twice, before and after administration and once daily on the other days.
- Body Weight: Body weight was measured before administration and 96 hours after administration.
- Necropsy: Not performed.
- Irritation:
The eyes of each animal were examined approximately 1, 24, 48, 72 and 96 hours after instillation of the test substance.
The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
0: No opacity
1: Scattered or diffuse area, details of iris clearly visible
2: Easily discernible translucent areas, details of iris slightly obscured
3: Opalescent areas, no details of iris visible, size of pupil barely discernible
4: Opaque, iris invisible

Area of cornea involved:
0: No opacity area
1: One quarter (or less) but not zero
2: Greater than one quarter, but less than half
3: Greater than half, but less than three quarters
4: Greater than three quarters, up to whole area

IRIS
0: Normal
1: Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof), iris still reacting to light (sluggish reaction is positive)
2: No reaction to light, hemorrhage, gross destruction (any or all of these)

CONJUNCTIVAL IRRITATION
Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris):
0: Vessels normal
1: Vessels definitely injected above normal
2: More diffuse, deeper crimson red, individual vessels not easily discernible
3: Diffuse beefy red

Chemosis:
0: No swelling
1: Any swelling above normal (includes nictitating membrane)
2: Obvious swelling with partial eversion of lids
3: Swelling with lids about half closed
4: Swelling with lids about half closed to completely closed

Discharge:
0: No discharge
1: Any amount different from normal (dose not include small amount observed in inner canthus of normal animals)
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs, and considerable area around the eye

Eye examinations were performed using a slit lamp.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

mean

Remarks:

Unwashed animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

Unwashed Animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

Unwashed Animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

3

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

Unwashed Animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

4

Results 5

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

Unwashed animal #1, #2, #3

Time point:

other: 1 hr

Score:

6

Max. score:

110

Reversibility:

fully reversible within: 24 hr

Results 6

Irritation parameter:

conjunctivae score

Remarks:

(opacity)

Basis:

mean

Remarks:

Washed animal #4, #5 and #6

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

4

Results 7

Irritation parameter:

iris score

Basis:

mean

Remarks:

Washed animal #4, #5 and #6

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

2

Results 8

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

Washed animal #4, #5 and #6

Time point:

other: 24, 28 and 72 hrs.

Score:

0

Max. score:

3

Results 9

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

Washed animal #4, #5 and #6

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

4

Results 10

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

Washed animal #4, #5, #6

Time point:

other: 1 hr

Score:

6

Max. score:

110

Reversibility:

fully reversible within: 24 hr

Irritant / corrosive response data:

Conjunctival redness, chemosis and discharge of score 1 were observed in all animals of eye-unwashed animals at 1 hour after administration. These changes disappeared by 24 hours after administration in all animals.

Individual and mean eye irritation scores are specified in the attached table.

Other effects:

Observation of the cornea using fluorescein sodium solution revealed no stain spot in any animal.

No clinical signs were observed in any animal throughout the observation period.

No abnormal body weight gain was observed in anyb animal throughout the observation period.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

An acute eye irritation/corrosion study with E-BW102 in the rabbit (6 males/dose) was conducted according to OECD 405 and GLP guidelines. Based on these study results, E-BW102 does not have to be classified and has no obligatory labelling requirement for eye irritation according to Regulation (EC) No. 1272/2008.