Sodium 1-amino-4-(cyclohexylamino)-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 26 to August 15, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 1-amino-4-(cyclohexylamino)-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

EC Number:

224-460-9

EC Name:

Sodium 1-amino-4-(cyclohexylamino)-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Cas Number:

4368-56-3

Molecular formula:

C20H219N2O5S.Na

IUPAC Name:

sodium 4-(4-acetamidophenyl-1-amino)-1-amino-9,10-dioxoanthracene-2-sulphonate

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Association of Breeders of Hradec Kralove.
- Weight at study initiation: above 2.5 Kg.
- Housing: animals were housed individually in metal cages without bedding with automatic air-condition.
- Diet: standard commercially manufactured feed mixture TM-Rabbit, ad libitum.
- Water: drinking water according to CSN 757111 ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3°C
- Humidity: 50 ± 15%
- Photoperiod: 12 hrs cycle dark/light
- Other: fluorescent light

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (g): 0.1 g

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours after instillation of the sample.

Number of animals or in vitro replicates:

3 animals (males and females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the eye was flushed with water.
- Time after start of exposure: 24 hrs.

TOOL USED TO ASSESS SCORE:
- Clinical examination of the left eye was performed by the use of ophthalmoscope Heine BETA 200 at intervals of 1, 24, 48 and 72 hours after instillation of the sample.
- At 24 hours, the damage of the corneal tissue was evaluated by the use of a 2% acqueous solution of fluorescein sodium salt.
- At 48 hours was the damage was evaluated without fluorescein.
- At 72 hours was the damage was evaluated by the use of a 2% acqueous solution of fluorescein sodium salt.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

ca. 1

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0.67

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Results 3

Irritation parameter:

iris score

Basis:

other: all animals

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

2

Reversibility:

fully reversible within: 24 hours

Results 4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

other: all animals

Time point:

24/48/72 h

Score:

ca. 2

Max. score:

3

Reversibility:

not fully reversible within: 72 hours

Results 5

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 6

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 1.33

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Results 7

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0.67

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

After 1 hour after application of the substance the following clinical signs are observed:
- Medial canthus: in rabbits, no. 1, no. 2 and no. 3 mucopurulent discharge, blepharospasm, narrowing of rima palpebrarum are observed.
- Bulbar and palpebral conjunctiva: rabbits, no. 1, no. 2 and no. 3 didn't show any signs of conjunctivitis. Swelling with partial lid inversion and chemosis is observed.
- Cornea: in rabbits, no. 1 and no. 2 opacity of cornea was observed; iris is not discernible. In rabbit, no. 3 the area is easily discernible translucent; details of iris slightly obscured
- Iris: in rabbits, no. 1, no. 2 and no. 3 there is total loss of light reaction.

After 24 hours after application of the substance the following clinical signs are observed:
- Medial canthus: in rabbits no. 1 and no. 3 mucopurulent medium discharge, blepharospasm and narrowing of rima palpebrarum. In rabbit, no. 2 large mucopurulent discharge, blepharospasm and narrowing of rima palpebrarum ae observed.
- Bulbar and palpebral conjunctiva: in rabbits, no. 1 no. 2 and no. 3 it is observed diffuse,crimson colour; individual vessels are not easily discernible. Difficult to evaluate the weak swelling
- Cornea: rabbits, no. 1, no. 2 and no. 3 diffuse areas of opacity; details of iris clearly visible
- Iris: rabbits, no. 1, no. 2 and no. 3 corresponded to physiological norms, without macroscopic pathomorfological changes

After 48 hours after application of the substance the following clinical signs are observed:
- Medial canthus: in rabbits, no. 1 and no. 3 there is central mucopurulent discharge, blepharospasm and narrowing of rima palpebrarum. Rabbit no. 2 shows a large mucopurulent discharge, blepharospasm and rima palpebrarum narrowing
- Bulbar and palpebral conjunctiva: in rabbits, no. 1, no. 2 and no. 3 it is observed diffuse,crimson colour; individual vessels are not easily discernible
- Cornea: in rabbits, no. 1 and no. 2 diffuse area of opacity, but details of iris are clearly visible. In rabbit, no. 3 cornea has a smooth, shiny appearance without macroscopic pathomorfologival changes.
- Iris: rabbits, no. 1, no. 2 and no. 3 corresponded to physiological norms, without macroscopic pathomorfological changes

After 72 hours after application of the substance the following clinical signs are observed:
- Medial canthus: in rabbits, no. 1 and no. 3 weak mucopurulent discharge. In rabbit no. 2 there is large mucopurulent discharge.
- Bulbar and palpebral conjunctiva: in rabbits, no. 1, no. 2 and no. 3 is observed diffuse,crimson colour; individual vessels are not easily discernible. Rabbit no.2 showed swelling with partial eversion of eye lid
- Cornea: in rabbits, no. 1 and No. 2 diffuse area of opacity, but details of iris are clearly visible
- Iris: rabbits, no. 1, no. 2 and no. 3 corresponded to physiological norms, without macroscopic pathomorfological changes

Applicant's summary and conclusion

Interpretation of results:

Category 2A (irritating to eyes) based on GHS criteria

Conclusions:

The test substance is irritating to eyes

Executive summary:

The substance was tested for Eye Irritation according to the OECD Guideline 405. The test substance (0.1 g) was placed as it is, in the conjuctival sac of the left eye after gently pulling the lower lid away from the eyeball. The lids were then held together for some seconds. The right eye was left untreated, and it served for control. For the assessement of eye irritation (in cornea, iris, conjuctivae) Three New Zealand white rabbits were used.

 

The eyes were evaluated and examined for any signs of occular lesions at 1, 24 ,48, 72 hours following test substance application. For the grading of the occular responses the harmonised scoring system as described in the OECD Guideline 405 was used. The response was calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material. The corneal opacity in at least 2 of 3 tested animals (animal #1 and animal #2) was equal to 1 and the conjuctival redness was equal to 2 in all the three animals.

 

The test substance is classified as Irritating to eyes (Category 2) according to the CLP Regulation (EC) No. 1272/2008.