Registration Dossier

Administrative data

Endpoint:
eye irritation, other
Remarks:
in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25-05-1981 to 02-07-1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study report which followed a known protocol (FDA/ Draize Method); acceptable restrictions.

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
1981

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: FDA of the United States (fed, reg, 28 (119), 5582, 1963)/ Draize and Kelley (Drug Cosmet, Industr, 71(1952) 36)
Principles of method if other than guideline:
In general the techniques of tests as published by the FDA of the United States (Fed. Reg. 28 (119), 5582, 1963) and Draize and Kelley (Drug Cosmet. Induster. 71 (1952) 36) are followed.

Six New Zealand White albino rabbits are used for each test substance. The animals are caged individually and received no hay or other extraneous material that might enter the eyes.

The eyes of the animals are examined before testing and only those animals without observable eye defects and used. One tenth of a milliliter of the test substance, or in the case of solids or aemisolids, 100 milligrams of the test substance, is allowed to fall on the everted lower lid of one eye of each rabbit; the upper and lower eye lid are then carefully closed and subsequently held together for at least one second before releasing, to prevent loss of material. The other eye, remaining untreated, serves as a control.

The eyes are not washed following instillation and the animals are released immediately.

An animal is considered as giving a positive reaction if there is, at any of the readings, discernible opacity of the cornea (other than a slight dulling of the normal lustre), or ulceration of the cornea, or inflammation of the iris (other than a slight deeping of the folds (rugae) or a slight circumcorneal injection), or if such substance produces in the conjunctivae (palpebral and bulbar, excluding the cornea and iris) an obvious swelling with partial eversion of the lids, or a diffuse deep-crimson red with individual vessels not easily discernible. The FDA-scoring scale is used.

The test is considered positive if four or more of the animals in the test group of six rabbits exhibit a positive reaction. If one animal exhibits a positive reaction, the test is regarded as negative.

If two or three animals exhibit a postive reaction, the test is repeated, using a different group of six animals. The second test is considered as positive if three or more of the animals exhibit a positive reaction. If only one or two animals in the second test exhibit a positive reaction, the test is again repeated with a different group of six animals. Should be a third test be needed, the substance will be regarded as irritant if two or more animals exhibit a positive response.
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: clear colourless liquid
Details on test material:
- Name of test material (as cited in study report): Ligustral U 05668
- Physical state: clear colourless liquid

Test animals / tissue source

Species:
rabbit
Strain:
other: New Zealand White albino rabbit

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): one tenth of a milliliter
Duration of treatment / exposure:
The treated eyes are not washed following instillation.
Observation period (in vivo):
24, 48, 72 hours and 7 days
Number of animals or in vitro replicates:
6 animals and unspecified sex
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: The method for grading ocular lesions is presented in Table 1 below.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: 1, 4 (individual mean)
Time point:
other: 24 h, 48h, 72h
Score:
0.67
Max. score:
1
Irritation parameter:
cornea opacity score
Basis:
animal: 2, 3, 5 (individual mean)
Time point:
other: 24h, 48 h, 72h
Score:
1
Max. score:
1
Irritation parameter:
cornea opacity score
Basis:
animal #6
Time point:
other: 24h, 48h, 72h
Score:
0
Max. score:
0
Irritation parameter:
iris score
Basis:
animal: 1, 2, 6 (individual mean)
Time point:
other: 24h, 48h, 72h
Score:
0.33
Max. score:
1
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: 24h, 48h, 72h
Score:
1
Max. score:
1
Irritation parameter:
iris score
Basis:
animal #4
Time point:
other: 24h, 48h, 72h
Score:
0
Max. score:
0
Irritation parameter:
iris score
Basis:
animal #5
Time point:
other: 24h, 48h, 72h
Score:
0.67
Max. score:
1
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 1, 2, 6 (individual mean)
Time point:
other: 24h, 48h, 72h
Score:
1.67
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #3
Time point:
other: 24h, 48h, 72h
Score:
2.33
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 4, 5 (individual mean)
Time point:
other: 24h, 48h, 72h
Score:
2
Max. score:
2
Irritation parameter:
chemosis score
Basis:
animal: 1, 5 (individual mean)
Time point:
other: 24h, 48h, 72h
Score:
1.67
Max. score:
2
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 24h, 48h, 72h
Score:
2
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 24h, 48h, 72h
Score:
2.33
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #4
Time point:
other: 24h, 48h, 72h
Score:
1.67
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #6
Time point:
other: 24h, 48h, 72h
Score:
1.33
Max. score:
2

Any other information on results incl. tables

Table 2: Individual scores awarded to the ocular lesions elicited by Ligustral

Rabbit number

Score

cornea iris Conjunctivae
 redness chemosis 
After 24 hours
1 1 1
2
3
4 0
After 48 hours
1
After 72 hours
1
2 1 0 1
After 7 days
1
5 0 0
0 0 0

 

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the FDA-standards, undiluted Ligustral is considered to be an eye irritant.
Executive summary:

In a primary eye irritation study (V81.203), 0.1 mL of undiluted Ligustral/Cyclal C/Trigustral was instilled into the eye lid of one eye of a New Zealand White albino rabbit (6 animals). The eye was not washed after instillation of the test article. Animals then were observed for 24 hr, 48 hr, 72 hr and 7 days after dosing. Irritation was scored using the system noted in Table 1.

The test substance caused slight corneal opacity in 5/6 rabbits, generally involving 50 -75% of the cornea, slight iritis also in 5/6 rabbits and moderate to severe redness and/or swelling of the conjunctivae in all animals. In addition five rabbits (no.1 through 5) showed moderate discharge of eye fluid. In the course of the seven-day observation period recovery of these lesions was observed but at the 7 -day reading one animal still showed slight corneal opacity, slight iritis and moderate conjunctivitis while two other animals showed slight redness of the conjunctivae.

In this study, Ligustral is an eye irritant based on the results of this test study according to FDA standards.