1,8-dihydroxy-4-nitro-5-(phenylamino)anthraquinone

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

None

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: 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).

Deviations:

no

Principles of method if other than guideline:

None

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

1,8-dihydroxy-4-nitro-5-(phenylamino)anthraquinone

EC Number:

243-632-4

EC Name:

1,8-dihydroxy-4-nitro-5-(phenylamino)anthraquinone

Cas Number:

20241-76-3

Molecular formula:

C20H12N2O6

IUPAC Name:

1-anilino-4,5-dihydroxy-8-nitro-9,10-anthraquinone

Specific details on test material used for the study:

None

Test animals / tissue source

Species:

rabbit

Strain:

other: Himalayan breed

Details on test animals or tissues and environmental conditions:

The test was performed on 3 male and 3 female rabbits of the Himalayan breed weighing 1.5 to 2 kg. The animals were housed individually in metal cages, were kept at a constant room temperature of 22 ± 1 °C, at a relative humidity of 55 ± 5 % and on a 10 hour light cycle day. The animals had ad libitum access to standard rabbit food - NAFAG, Gossau SG - and water. Prior to treatment they were adapted to our laboratories for a minimum of 4 days. Only rabbits with normal ophthalmic findings were used for these tests.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The right eye was not treated and served as an untreated control.

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 ml of lukewarm water. 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:

6 (3 males and 3 females)

Details on study design:

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. 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.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

eyes not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

4

Results 2

Irritation parameter:

cornea opacity score

Remarks:

eyes rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

1.22

Max. score:

4

Reversibility:

fully reversible within: 7d

Results 3

Irritation parameter:

iris score

Remarks:

eyes rinsed and not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 4

Irritation parameter:

conjunctivae score

Remarks:

eyes rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

2.67

Max. score:

3

Reversibility:

fully reversible within: 7d

Results 5

Irritation parameter:

chemosis score

Remarks:

eyes rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

1.89

Max. score:

4

Reversibility:

fully reversible within: 7d

Results 6

Irritation parameter:

conjunctivae score

Remarks:

eyes not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

2.33

Max. score:

3

Reversibility:

fully reversible within: 7d

Results 7

Irritation parameter:

chemosis score

Remarks:

eyes not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

1.6

Max. score:

4

Reversibility:

fully reversible within: 7d

Irritant / corrosive response data:

In the present experiments the irritation index was established as 3.9 for the cornea, 0 for the iris and 6 for the conjunctivae. The test material caused therefore a minimal overall eye irritation to rabbits.
Rinsing the eyes following instillation was of practically no effect.

Any other information on results incl. tables

Table 1: Eye irritation in individual rabbits with eyes rinsed 30 sec. after treatment

Days after Appl.	Rabbit 1 (MALE)			Rabbit 2 (MALE)			Rabbit 3 (MALE)
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
1	10	0	10	5	0	10	5	0	10
2	10	0	10	5	0	8	5	0	6
3	10	0	8	0	0	6	5	0	0
4	5	0	6	0	0	0	0	0	0
7	0	0	0	0	0	0	0	0	0

Cornea: 4; Iris: 0; Conjunctivae: 5.6

Table 2: Eye irritation in individual rabbits with eyes not rinsed after treatment

Days after Appl.	Rabbit 4 (FEMALE)			Rabbit 5 (FEMALE)			Rabbit 6 (FEMALE)
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
1	5	0	10	5	0	12	5	0	10
2	5	0	10	5	0	10	5	0	10
3	5	0	8	5	0	8	5	0	6
4	5	0	8	0	0	6	0	0	0
7	5	0	0	0	0	0	0	0	0

Cornea: 3.7; Iris: 0; Conjunctivae: 6.5

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

FAT 36080/A can be considered as a mild eye irritant.

Executive summary:

The eye irritation potential of FAT 36080/A 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. 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 (A) and rinsed (B) eyes respectively. The irritation index was established as 3.9 for the cornea, 0 for the iris and 6 for the conjunctivae. The test material caused therefore a minimal overall eye irritation to rabbits. Rinsing the eyes following instillation was of practically no effect. The scores obtained in the study meet the threshold scores proposed by CLP Regulation for classification as eye irritant category 2. The mean scores for corneal opacity and conjunctival redness were scored as ≥1 and ≥2 respectively, for at least 2 out of 3 animals which received rinsing. Hence, FAT 36080/A can be classified as eye irritant category 2 according to the CLP (Regulation EC No. 1272/2008) criteria for classification.