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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:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented, according to accepted guidelines.

Data source

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

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
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
GLP compliance:
yes
Remarks:
Statement of GLP not signed.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): diethyl ether.
- Physical state: Liquid (clear).
- Analytical purity: 99.93 %.
- Lot/batch No.: 09023198.
- Expiration date of the lot/batch: 09-2010 (as stated by the Sponsor); 30-Sep-2010 (as handled by Harlan Laboratories Ltd.).
- Storage condition of test material: At room temperature (range of 20 ± 5 °C, provided by Harlan Laboratories Ltd.), light protected.

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd., Hillcrest, Dodgeford Lane, Belton, Loughborough, Leics, LE12 9TE / UK.
- Age at study initiation: 11 weeks.
- Weight at study initiation: 2472 - 2571 g.
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls.
- Diet (e.g. ad libitum): Pelleted standard Teklad Global High Fiber Rabbit Diet 2031C (batch no. 25/10, Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) was available ad libitum.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 23-Sep-2010 to 27-Sep-2010 (one animal); 23-Sep-2010 to 28-Sep-2010 (two animals).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 °C.
- Humidity (%): 30-70%.
- Air changes (per hr): 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 / 12.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL/animal.
Duration of treatment / exposure:
Single exposure
Observation period (in vivo):
1, 24, 48, and 72 hours after administration.
Number of animals or in vitro replicates:
3 males.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data.

SCORING SYSTEM: The eye reactions were assessed according to the numerical scoring system listed in the Commission Regulation (EC) No 440/2008, B.5, at approximately 1, 24, 48 and 72 hours after administration. Scleral reddening and ocular discharge were also assessed.

Commission Regulation (EC) No 440/ 2008, 30 May 2008 - Grading of Ocular Lesions.

1. 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 luster), details of iris clearly visible: 1
Easily discernible translucent area, details of iris slightly obscured: 2
Nacreous area, no details of iris visible, size of pupil barely discernible: 3
Opaque cornea, iris not discernible through the opacity: 4

2. Area of cornea involved
Zero: 0
One quarter (or less) but not zero: 1
Greater than one quarter, but less than half: 2
Greater than half, but less than three quarters: 3
Greater than three quarters, up to whole area: 4

3. Iris
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive): 1
No reaction to light, hemorrhage, gross destruction (any or all of these): 2

4. Conjunctivae
-Redness (refers to most severe reading of palpebral and bulbar conjunctivae when compared with control eye)
Blood vessels normal: 0
Some blood vessels definitely hyperemic (injected): 1
Diffuse, crimson color, individual vessels not easily discernible: 2
Diffuse beefy red: 3

-Chemosis: lids and/or nictitating membranes
No swelling: 0
Any swelling above normal (including nictitating 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

Additionally, ocular discharge, reddening of the sclerae and staining of conjunctivae, sclerae and cornea by the test item were assessed according to the following scheme:

1. Ocular Discharge
No discharge: 0
Slight: Any amount different to normal (does not include small amount observed in inner canthus of normal animal): 1
Moderate: Discharge with moistening of the lids and hair just adjacent to the lids: 2
Marked: Discharge with moistening of the lids and hairs, and a considerable area around the eye (running): 3

2. Sclerae
-Redness
Normal: Blood vessels normal: 0
Slight reddening: Some blood vessels definitely hyperemic (injected): 1
Moderate reddening: Diffuse, crimson color, individual vessels not easily discernible: 2
Marked reddening: Diffuse beefy red: 3

3. Staining of Conjunctivae, Sclerae and Cornea by the Test Item
Not observed: 0
Slight staining: 1
Marked staining: 2

The test item was classified according to the referred classification criteria: Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labeling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and
1999/45/EC, and amending Regulation (EC) No 1907/2006.

TOOL USED TO ASSESS SCORE: Eye examinations were made with a Varta Cliptrix diagnostic-lamp (Roth AG, 4153 Reinach / Switzerland).

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Remarks:
-Corneal opacity and area of corneal opacity
Basis:
mean
Remarks:
-3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
mean
Remarks:
-3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
-Redness
Basis:
animal #1
Time point:
other: 1 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Remarks:
-Redness
Basis:
mean
Remarks:
-Animals 2 and 3
Time point:
other: 1 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 48 h
Irritation parameter:
conjunctivae score
Remarks:
-Redness
Basis:
mean
Remarks:
-Animals 1 and 2
Time point:
24 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Remarks:
-Redness
Basis:
animal #3
Time point:
24 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 48 h
Irritation parameter:
conjunctivae score
Remarks:
-Redness
Basis:
animal #1
Time point:
48 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Remarks:
-Redness
Basis:
mean
Remarks:
- Animals 2 and 3
Time point:
48 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 48 h
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
-3 animals
Time point:
other: 1 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 24 h
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
-3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 24 h
Irritation parameter:
other: Sclera
Basis:
animal #1
Time point:
other: 1 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 24 h
Irritation parameter:
other: Sclera
Basis:
mean
Remarks:
-Animals 2 and 3
Time point:
other: 1 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 24 h
Irritation parameter:
other: Sclera
Basis:
mean
Remarks:
-3 animals
Time point:
other: 24, 48, and 72 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 h
Irritation parameter:
other: Ocular discharge
Basis:
mean
Remarks:
-Animals 1 and 3
Time point:
other: 1 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 24 h
Irritation parameter:
other: Ocular discharge
Basis:
animal #2
Time point:
other: 1 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 h
Irritation parameter:
other: Ocular discharge
Basis:
mean
Remarks:
-3 animals
Time point:
other: 24, 48, and 72 h
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 24 h
Irritation parameter:
conjunctivae score
Remarks:
- Redness
Basis:
mean
Remarks:
3 animals
Time point:
72 h
Score:
0
Max. score:
3
Reversibility:
fully reversible
Irritant / corrosive response data:
1. Irritation:
The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iris light reflex, redness and chemosis of the conjunctivae, respectively. The individual mean scores for corneal opacity and iris light reflex were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.67, 0.33 and 0.00 for reddening and 0.00 for chemosis for all three animals. Slight to moderate reddening of the conjunctivae, slight to moderate reddening of the sclerae and slight swelling (chemosis) of the conjunctivae were noted in all three males 1 hour after instillation. The reddening of the conjunctivae persisted as slight in two animals (Nos. 87 and 88) up to the 24- or 48-hour reading. Slight ocular discharge was present in two animals (Nos. 87 and 89) at the 1-hour observation.
No abnormal findings were observed in the treated eyes of any animals 72 hours after treatment, the end of the observation period for all animals.

2. Coloration:
No staining produced by the test item was observed in the treated eyes.

3. Corrosion:
No corrosion of the cornea was observed at any of the reading times.

Refer to Table 1.
Other effects:
1. Viability / Mortality: No intercurrent deaths occurred during the course of the study.
2. Clinical Signs: No clinical signs were recorded throughout the entire observation period.
3. Body Weights: The body weight of the animals was within the range commonly recorded for this strain and age.
4. Macroscopic Findings: No necropsy was performed at the end of the study.

Any other information on results incl. tables

Table 1: Eye Irritation Scores – Mean Values after 24, 48, and 72 hours

Animal number

Sex

Corneal Opacity

Iris Light Reflex

Conjunctivae

Redness

Chemosis

87

Male

0.00

0.00

0.67

0.00

88

Male

0.00

0.00

0.33

0.00

89

Male

0.00

0.00

0.00

0.00

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Remarks:
Criteria used for interpretation of results: other: CLP (EC 1272/2008)