Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
6 March to 15 March 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study to GLP conducted using material that consisted of 65 % active ingredient and 35 % xylene.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Benzoic acid, 2-hydroxy-,C14-18 alkyl derivs.
EC Number:
931-472-4
Cas Number:
182700-89-6
IUPAC Name:
Benzoic acid, 2-hydroxy-,C14-18 alkyl derivs.
Constituent 2
Reference substance name:
605-976-7
EC Number:
605-976-7
IUPAC Name:
605-976-7
Test material form:
liquid: viscous
Details on test material:
- Physical state: pale brown viscous liquid
- Stability under test conditions: no data
- Storage condition of test material: under nitrogen at ambient temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Age at study initiation: about 5 months
- Weight at study initiation: 3.44 to 3.78 kg
- Housing: individually, in suspended stainless steel cages
- Diet (e.g. ad libitum): conventional, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 1 to 2 months

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 22
- Humidity (%): 32 to 57
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
- Rabbit No.1: From 6 March 1995 to 13 March 1995
- Rabbits Nos. 2 and 3: From 8 March 1995 to 15 March 1995

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
Duration of treatment / exposure:
single application, not washed out of the eye
Observation period (in vivo):
8 days
Number of animals or in vitro replicates:
Three. One animal was initially treated with the test material. As a severe irritation response was not observed, two more rabbits were treated.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: scored according to OECD guideline. Also scored for pain reaction to instillation of the test material and for area of the cornea affected by the lesion and discharge from the conjunctivae.

TOOL USED TO ASSESS SCORE: ophthalmoscope or pencil beam torch. From 24 h fluorescein was used for examination of the cornea, viewed under uv-light.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: mean 24, 48 and 72 h
Score:
0.8
Max. score:
4
Reversibility:
other: reversible in 2/3 rabbits within 48 or 72 h
Irritation parameter:
iris score
Basis:
mean
Time point:
other: mean 24, 48 and 72 h
Score:
0.7
Max. score:
2
Reversibility:
other: fully reversible in 2/3 rabbits within 48 h
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: mean 24, 48 and 72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: mean 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
other: fully reversible in 2/3 rabbits in 48 or 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: no reaction
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: mean 24, 48 and 72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: mean 24, 48 and 72 h
Score:
1.7
Max. score:
4
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 h
Score:
0.3
Max. score:
2
Reversibility:
fully reversible within: 48 h
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: mean 24, 48 and 72 h
Score:
0.7
Max. score:
2
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: mean 24, 48 and 72 h
Score:
1
Max. score:
2
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: mean 24, 48 and 72 h
Score:
1.7
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: mean 24, 48 and 72 h
Score:
2.3
Max. score:
3
Reversibility:
fully reversible within: 8 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: mean 24, 48 and 72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: mean 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: mean 24, 48 and 72 h
Score:
1.7
Max. score:
4
Reversibility:
not fully reversible within: 8 days
Irritant / corrosive response data:
The sentinel rabbit (No.1) showed a diffuse beefy-red conjunctiva during the first 24 h, which gradually reduced but injection of the blood vessels persisted to study termination on day 8. Iritis, very slight chemosis and slight to moderate discharge with mucus were evident during the first 24 h but had reversed by 48 h, 48 h and 8 days, respectively. No effects on the cornea were apparent.

Animal No.2 showed pink to crimson conjunctiva and very slight or slight discharge (with mucus) during the first 72 h but had mainly reversed by day 8. An area of very slight corneal opacity, very slight or slight chemosis and iritis were seen at 24 and 48 h.

Animal No.3 showed crimson or beefy-red conjunctiva, very slight or slight chemosis, very slight or slight discharge, iritis and opacity of the cornea all of which were still present on day 8. Pannus formation (which is irreversible) affecting both the upper and lower edges of the cornea was evident on day 8, which resulted in the decision to terminate the study at this point.
Other effects:
The pain response for animal No. 1 upon instillation of the test substance was moderate (grade 3; on a scale of 0 to 5).

Applicant's summary and conclusion

Interpretation of results:
Category I
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material was irritating when 0.1 mL was instilled into the eyes of three New Zealand White rabbits.
Executive summary:

In a GLP study conducted according to OECD Guideline 405 and to EU Method B.5, the test material (approximately 65 % active substance, 35 % xylene) was assessed for its irritant potential when instilled into the eyes of three New Zealand White rabbits.

Each animal was treated with 0.1 mL of the undiluted test material in the right eye and the eyelids gently held together for 1 second before releasing. The left eye was untreated and served as a concurrent control.The rabbits were observed at 1, 24, 48 and 72 h and at 8 days after instillation for signs of occular irritation. Initially only one animal was treated which experienced a moderate pain response; to prevent this in the other two animals they were treated with an anesthetic before instillation of the test material (including the control eye).

Moderate to severe conjunctivitis was evident in all three rabbits during the first 72 h after exposure which was not fully reversible by 8 days. Moderate iritis was seen in one animal at 1 and 24 h, at 24 and 48 h in another, and throughout the study in the third. No corneal opacity was evident in one animal, was present at 24 and 48 h in a second and in the third was present from 1 h through to day 8. In this latter animal the irreversible lesion, pannus, was evident on day 8, affecting both the upper and lower edges of the cornea.

In conclusion, the test material was irritating when 0.1 mL was instilled into the eyes of three New Zealand White rabbits, causing pannus in one animal.