Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
see - comment below
Principles of method if other than guideline:
No music was played during the light cycle some days during the housing of the animals.
The treated eye of the animals was first rinsed with bi-distilled water and immediately afterwards with saline 0.9 %.
The relative humiity was observed below 40 % in the test laboratory.
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
prop-2-en-1-yl 2-(2-chloro-5-nitrobenzoyloxy)-2-methylpropanoate
EC Number:
696-317-2
Cas Number:
174489-76-0
Molecular formula:
C14H14ClNO6
IUPAC Name:
prop-2-en-1-yl 2-(2-chloro-5-nitrobenzoyloxy)-2-methylpropanoate

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne / France
- Age at study initiation: 15 weeks (male, 14 weeks (females)
- Weight at study initiation: 2.7 - 3.2 kg
- Housing: Indiviadual
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 28 - 70 %
- Air changes (per hr): 10 - 15/hour
- Photoperiod (hrs dark / hrs light): 12/12 hour

Test system

Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied : 0.1 g
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
1, 24, 48 and 72 days (scored for lesions). Also, 7, 10 14 and 17 days.
Number of animals or in vitro replicates:
3 (1 male, 2 females)
Details on study design:
APPLICATION OF TEST SUBSTANCE:
The eyes of the animals were examined one day prior to test item administration.
On the day of treatment, 0.1 g of the test item was place d in the conjunctival sac of the left eye of each animal. The right eye remained untreated and served as the reference control.

REMOVAL OF TEST SUBSTANCE:
After 24 hour reading, the treated eye of each animal was rinsed with bi-distilled water until no remnants were left (60 ml syringe with an infusion cannula (round shaped top), approx. 150 ml bi-distilled water). Immediately after this procedure, the eye of each animal was then additionally rinsed with saline 0.9 % (approx. 20 ml).

OBSERVATIONS:
Viability/Mortality: Daily from delivery of the anmals to the termination of test.
Clinical signs: Daily from delivery of the anmals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation.

SCORING SYSTEM:
The ocular reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 at
approximately 1, 24, 48, 72 hours, as well as 7, 10, 14 and 17 days after administration.

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 1 hour
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible within: 10 days
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24 hours
Score:
ca. 1
Max. score:
2
Reversibility:
fully reversible within: 10 days
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 48 hours
Score:
ca. 1
Max. score:
2
Reversibility:
fully reversible within: 10 days
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 72 hours
Score:
ca. 1
Max. score:
2
Reversibility:
fully reversible within: 10 days
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 1 hour
Score:
ca. 0
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: 2 of 3 results not visible due to conjunctival swelling.
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24 hours
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: all 3 results not visible due to conjunctival swelling.
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 48 hours
Score:
ca. 0
Max. score:
0
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 72 hours
Score:
ca. 0
Max. score:
0
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 1 hour
Score:
ca. 1
Max. score:
2
Reversibility:
fully reversible within: 17 days
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24 hours
Score:
ca. 2
Max. score:
3
Reversibility:
fully reversible within: 17 days
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 48 hours
Score:
ca. 2
Max. score:
3
Reversibility:
fully reversible within: 17 days
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 72 hours
Score:
ca. 2
Max. score:
2
Reversibility:
fully reversible within: 17 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 1 hour
Score:
ca. 3
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24 hours
Score:
ca. 4
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 48 hours
Score:
ca. 2
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 72 hours
Score:
ca. 1
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
Slight to moderate corneal opacity was seen in all animals at the 24 hour reading and persisted through 72 hours with more or less severity. Diffuse areas of opacity were seen I only one animal at the 7 day reading before disappearing by day 10.

Slight to moderate redness of the conjunctivae and nictitating membrane was observed in all animals at the 1 hour examination and increased in severity at 24 hours beforediminishing to clear by day 10 (two animals) or 17 (one animal). Marked swelling of the conjunctivae and nicitiating membrane with lids about half closed was observed at 1 hour and increased to swelling with the lids more than half closed after 24hours.

Due to marked conjunctival swelling, the iris could not be observed from 1 hour (two animals) to 24 hours (all animals) after treatment. During the remaining observation period, no abnormal effect on the iris was observed. The sclera was also not visible in all animals at 1 and 24 hours due to the swelling. Moderately to markedly reddened sclera was seen in two animals at 48 hours, staying visible in the third animal before disappearing by day 7 (two animals) or 14 (one anima). Slight to marked (watery or mucous) discharge was observed in all animals at the 1 hour reading which increased to marked discharge in all animals at 24 hours before diminishing to clear by 48 hours (one animal) or 72 hours (two animals). All eye reactions were clear within 17 days after treatment.

The mean values from 24 to 72 hours were 1.3 for corneal opacity, 0 for iris, 2.2 for conjunctival redness and 2.6 for conjunctival chemosis.
Other effects:
No clinical signs of systemic toxicity were observed in the animals during the test and observation period and no mortality occurred.

No staining of the treated eyes by the test item was observed.

Yellowish remnants of the test item were observed in eye or conjunctival sac of all animals at 1 and 24 hour examinations. The remnants were removed during the rinsing of the treated eyes after 24 hour reading.

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

The body weight of all rabbits were considered to be within the normal range of variability.

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information to eyes Criteria used for interpretation of results: EU
Conclusions:
Based upon the criteria stated in the EEC Commission Directive 93/21/EEC, CA 2218 A (Intermediate of CGA 276854) is required to be classified as ‘Irritating to eyes’.
Executive summary:

The primary eye irritation potential of CA 2218 A (Intermediate of CGA 276854) was investigated by installation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. The treated eyes were rinsed just after the 24 hour examination. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14 and 17 days after test application. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion. 

 

Marked swelling of the conjunctivae and nictitating membrane with lids about half-closed was observed at the 1 hour and increased in severity by 24 hours. The treated eyes of the animals were rinsed after the 24 hour reading; the severity of the swelling decreased in all animals at 48 hours before disappearing by day 7. 

 

Slight to moderate corneal opacity was seen in all animals at the 24 hour reading and persisted through 72 hours in two animals and through the 7 day reading in the third. Due to marked conjunctival swelling, the iris could not be observed from 1 hour (two animals) to 24 hours (all animals) after treatment. During the remaining observation period, no abnormal effect on the iris was observed. Slight to moderate redness of the conjunctivae and nictitating membrane was observed in all animals at the 1 hour examination and increased in severity at 24 hours before diminishing to clear by day 10 (two animals) or 17 (one animal). 

 

The sclera was also not visible in all animals at 1 and 24 hours and in one animal t 48 hours due to the swelling. Moderately to markedly reddened sclera was seen in two animals at 48 hours and in all three animals at 72 hours, before disappearing by day 7 (two animals) or 14 (one animal). Slight to marked discharge was observed in all animals at the 1 hour reading and increased in severity at 24 hours before diminishing to clear by 48 hours (one animal) or 72 hours (two animals). All eye reactions were clear within 17 days after treatment. 

 

 

The mean values from 24 to 72 hours were 1.3 for corneal opacity, 0 for iris, 2.2 for conjunctival redness and 2.6 for conjunctival chemosis.

 

Based upon the criteria stated in the EEC Commission Directive 93/21/EEC, CA 2218 A (Intermediate of CGA 276854) is required to be classified as ‘Irritating to eyes’.