Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
9 May to 18 May, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test method according to EU method B.5. GLP study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
Eye irritation
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): OS 2200 (489-95A)
- Physical state: Clear to light yellow liquid.
- Analytical purity: >92%.
- Lot/batch No.:38659-14
- Expiration date of the lot/batch:1 January 1996.
- Storage condition of test material: Room temperature in the dark under nitrogen.

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Froxfield (U.K.) Ltd., Petersfield, Hampshire, England.
- Age at study initiation: 12 to 14 weeks.
- Weight at study initiation: 2.8 to 3.2kg.
- Housing: individually housed in plastic cages with perforated floors.
- Diet (e.g. ad libitum): Ad libitum. Standard laboratory diet SDS Stanrab (P) rabbit diet.
- Water (e.g. ad libitum): Ad libitum.
- Acclimation period: All rabbits were acclimatised to the experimental environment.

ENVIRONMENTAL CONDITIONS
- Temperature : 19ºC.
- Humidity : 30-70%.
- Air changes (per hr): 19 per hour.
- Photoperiod (hrs dark / hrs light): 12hours dark/12 hours artificial light.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1mL/eye/rabbit.
Duration of treatment / exposure:
The eyelids were gently held together for one second before releasing.
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: After instillation, the eyelids were gently held together for one second before releasing

SCORING SYSTEM:
Ocular irritation was assessed using the prescribed numerical system:
Cornea
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Nacreous areas, no details of iris visible, size of pupil barely discernible 3
Opaque cornea. iris not discernible through the opacity 4
Iris
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, haemorrhage, gross destruction (any or all of these) 2

Conjunctivae
Redness (refers to the most severe reading of palpebral and bulbar conjunctivae, as compared to the control eye)
Blood vessels normal 0
Some blood vessels definitely hyperaemic (injected) 1
Diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3
Chemosis (lids and/or nictating membranes)
No swelling 0
Any swelling above normal (includes nictating membranes) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half-closed 3
Swelling with lids more than half-closed 4

TOOL USED TO ASSESS SCORE: Hand-slit lamp.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 24-72h
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 3 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 24-72h
Score:
0
Max. score:
0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
other: 24-72h
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 3 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 24-72h
Score:
0.3
Max. score:
1
Reversibility:
fully reversible within: 2 days
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: 24-72h
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 3 days
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: 24-72h
Score:
0
Max. score:
0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Time point:
other: 24-72h
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 3 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 24-72h
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 3 days
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: 24-72h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: 24-72h
Score:
0
Max. score:
0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #3
Time point:
other: 24-72h
Score:
1.3
Max. score:
2
Reversibility:
fully reversible within: 4 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 24-72h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 3 days
Irritant / corrosive response data:
Corneal opacification was seen in all three animals. Stripping of the epithelial cells from the corneal surface was seen in one animal one hour after, dosing only. No iridial inflammation was observed. A diffuse crimson colouration of the conjunctiva was seen in all three animals and was accompanied in one animal by obvious swelling with partial eversion of the eyelids. The eyes were normal 3 or 7 days after instillation.
Other effects:
There were no signs of toxicity or ill health in any rabbit during the observation period.

Any other information on results incl. tables

Table 1. Ocular reactions observed after instillation of OS-2200

Time after instillation

Day after instillation

Rabbit

Region of eye

1 hour

1

2

3

4

7

816*

Cornea

D

1

1

0

0

0

Iris

0

0

0

0

0

0

Conjunctiva

Redness

2

1

1

0

0

0

Chemosis

1

1

0

0

0

0

817

Cornea

aD

1

1

0

0

0

Iris

0

0

0

0

0

0

Conjunctiva

Redness

2

1

1

0

0

0

Chemosis

1

1

1

0

0

0

800

Cornea

D

1

1

1

1

0

Iris

0

0

0

0

0

0

Conjunctiva

Redness

2

2

1

1

0

0

Chemosis

1

2

1

0

0

0

*: Pilot animal

a Stripping of epithelial cells from the corneal surface

D Dulling

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The substance was determined to be not an irritant to the eye.
Executive summary:

An ocular irritation study was performed according to EU method B.5 (GLP study). Eye irritation was assessed by instilling 0.1mL of OS-2200 into one eye of three rabbits. The contralateral eye served as control. The eyes were observed for signs of irritation for 7 days after exposure. Corneal opacity was observed in all three animals. No irritation of the iris occurred. A diffuse crimson discoloration of the tissue surrounding the eye and lining the eyelids (conjunctiva) was noted in all three rabbits. This was accompanied by swelling and partial eversion of the eyelids in one animal. These effects lasted from 3 to 7 days. The test item was determined to be not irritating to eyes according to CLP Regulation.