2-(6-methoxybenzofuran-2-yl)-1,3-dimethyl-5-(methylsulphonyl)1H-benzimidazolium acetate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Principles of method if other than guideline:

As per the procedure described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO).

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Fluorescent Brightener 363

IUPAC Name:

Fluorescent Brightener 363

Test animals / tissue source

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.5 to 2 kg.
- Housing: animals were housed individually in metal cages.
- Diet: rabbit food - NAFAG, Gossau SG -, ad libitum.
- Water: ad libitum.
- Acclimation period: animals were adapted to our laboratories for a minimum of 8 days. Only rabbits with normal ophthalmic findings were used for these tests.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1 °C
- Relative humidity: 55 ± 5 %
- Photoperiod: 14 hours light cycle day.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

In 3 out of 6 rabbits treated eye was flushed approximately 30 seconds after treatment; the remaining 3 rabbits did not receive any flushing during the entire study period.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 male and 3 female rabbits

Details on study design:

APPLICATION OF TEST SUBSTANCE
The test material in an amount of 0.1 g was inserted into the conjunctival sac of the left eye of the 6 rabbits and the lids were gently closed for one second. The right eye was not treated and served as an untreated control.

REMOVAL OF TEST SUBSTANCE
- Washing: in 3 out of the 6 rabbits the treated eye was flushed with the with 10 ml of lukewarm water.
- Time after start of exposure: approximately 30 seconds after treatment .

TOOL USED TO ASSESS SCORE: hand-slit lamp.

OBSERVATION TIME POINTS
The eye irritation was appraised with a slit-lamp on day 1, 2, 3, 4 and 7.

SCORING SYSTEM
The eye irritation was scored for each individual rabbit. The primary eye irritation, index represents the sum of the mean values for cornea, iris and conjunctivae. This procedure was carried out separately for unrinsed and rinsed eyes respectively.

Score for eye irritation in rabbits:
Cornea
A Opacity and degree of density (most dense area scored)
No opacity 0
Scattered or diffuse area, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris invisible 4

B Area of cornea involved
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

A x B x 5
Maximum possible score = 80

Iris
A Values
Normal 0
Folds above normal, congestion, swelling, circumcornea injection (any or all 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

A x 5
Maximum possible score = 10

Conjunctivae
A Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

B Chemosis
No swelling 0
Any swelling above normal (includes nictitating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids about half closed to completely closed 4

C Discharge
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adj acent to lids 2
Discharge with moistening of the lids and hairs, and considerable area around the eye 3

(A + B + C) x 2
Maximum possible score = 20

The total maximum score was thus 80 for the cornea, 10 for the iris, 20 for the conjunctivae and 110 for the entire eye. The severity of overall eye irritation was rated as:
0 - 10 minimal
11 - 25 slight
26 - 56 moderate
57 - 84 marked
above 84 extreme

The ratio unrinsed Vs rinsed eyes indicates the effect of rinsing the eyes 30 seconds after instillation of the compound.
<1 = rinsing increased the effect of the compound
1.1-1.5 = practically no effect
R = 1.6-2 = little but assessable effect
2.1-4 = good effect
4.1-10 = very good effect
>10 = action of the compound practically abolished

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 2/3

Remarks:

rinsed eyes

Time point:

24/48/72 h

Score:

>= 1

Reversibility:

fully reversible within: 4 days

Results 2

Irritation parameter:

iris score

Basis:

animal: 3/3

Remarks:

rinsed eyes

Time point:

24/48/72 h

Score:

< 1

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Remarks:

rinsed eyes

Time point:

24/48/72 h

Score:

< 2

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Remarks:

rinsed eyes

Time point:

24/48/72 h

Score:

< 2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The irritation index was established as 3.4 for the cornea, 0 for the iris and 1.6 for the conjunctivae.
The test material caused therefore a minimal overall eye irritation to rabbits.
Effect of rinsing was determined to be 3.1, indicating the 'good effect' of rinsing in minimising the irriatation effect of the test substance.

Any other information on results incl. tables

Animal N./sex	Effect	Days after application
		1	2	3	4	7
RINSED EYES
1 F	Cornea	5	5	0	0	0
2 F	Cornea	5	5	5	0	0
3 F	Cornea	5	5	5	0	0
1 F	Iris	0	0	0	0	0
2 F	Iris	0	0	0	0	0
3 F	Iris	0	0	0	0	0
1 F	Conjuntivae	0	0	0	0	0
2 F	Conjuntivae	0	0	0	0	0
3 F	Conjuntivae	0	0	0	0	0
NOT RINSED EYES
4 M	Cornea	10	10	5	0	0
5 M	Cornea	10	5	5	0	0
6 M	Cornea	10	5	5	0	0
4 M	Iris	0	0	0	0	0
5 M	Iris	0	0	0	0	0
6 M	Iris	0	0	0	0	0
4 M	Conjuntivae	8	10	6	0	0
5 M	Conjuntivae	6	6	0	0	0
6 M	Conjuntivae	6	6	0	0	0

Applicant's summary and conclusion

Interpretation of results:

other: eye irrit 2 (H319), according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Minimal eye irritating

Executive summary:

The eye irritation potential of the test substance was evaluated in a study conducted as per the procedure described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO). The test was performed on 3 male and 3 female rabbits of Himalayan breed weighing 1.5 to 2 kgs. The test material in an amount of 0.1 g was inserted into the conjunctival sac of the left eye of the 6 rabbits and the lids were gently closed for one second. The right eye was not treated and served as an untreated control. In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 ml of lukewarm water. The eye irritation was appraised with a slit-lamp on day 1, 2, 3, 4 and 7 and was scored for each individual rabbit.

A slight persistent blue coloration of the sclera was observed throughout the test. The irritation index was established as 3.4 for the cornea, 0 for the iris and 1.6 for the conjunctivae. The test material caused therefore a minimal overall eye irritation to rabbits. Rinsing the eyes following instillation was found to be of ‘good effect’ in minimizing the irritation.

Conclusion

Although the criteria for the evaluation of the eye irritation potential followed in the test are different from those reported into the OECD guideline currently in force, the scoring system used for corea opacity and degree of density, for the iris and for the conjunctivae redness and chemosis is the same of that reported into the OECD. Thus, because the raw tables are included into the study report, it is possible to re-evaluate the results according to the CLP Regulation (EC) 1272/2008.

The mean values from gradings at 24, 48 and 72 hours for rinsed eyes were equal to 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in at least 2 out of 3 tested animals.

Therefore the substance meets the criteria to be classified as eye irritating, according to the CLP (EC) 1272/2008 Regulation.