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

Endpoint summary

Description of key information

Skin Irritation: Non irritating
Eye Irritation: Irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 26 to August 15, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Association of Breeders of Hradec Kralove.
- Weight at study initiation: above 2.5 Kg.
- Housing: rabbits were housed invidually in metal cages commercially produced without bedding, with automatic air-condition.
- Diet: rabbits were fed standard commercially manufacturing 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

Type of coverage:

occlusive

Preparation of test site:

other: shaved and abraded

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied : 0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: an area of 6 cm x 6 cm was depilated.
- Type of wrap if used: the sample was covered with gauze, above which aluminum film and cellulose wadding was applied. Technical tape was used so that the test substance was in contact with the depilated skin.

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 4 hours.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

The mean erythema and edema score for the three rabbits is 0.

At 4, 24, 48 and 72 hours after removal of patch no rmacroscopic or pathomorphological changes were identified on the intact or scarified skin. The present sample after epicutaneous application to the skin of rabbits did not induce contact dermatitis.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material is not irritant to the rabbits

Executive summary:

The substance was evaluated for Skin irritation/Corossion according to the OECD Guideline 404. The test substance (0.5 g) after the treatment with water was applied as a paste on the intact (left lateral depilated area) and scarified skin (right lateral depilated area) of Three New Zealand white rabbits. Depilated area of skin outside the application zone was used to control the assessment.

 

After 4 - 24 - 48 - 72 hours of the removal of the patch the animals were examined for signs of erythema and oedema. No edema or erythema signs were identified on the intact skin and neither on the scarified skin.

 

Based on erythema and edema scores, the test substance is not considered to be irritating to the skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

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

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.

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

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

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

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

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

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

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

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation/Corossion

The test substance was evaluated in the key study for its skin irritation potential according to the OECD guideline 404.

0.5 g of the test substance was applied on the skin of three rabbits using gauze, then covered by aluminium foil and cellulose wadding. After 4 hours the gauze was removed and the skin was observed for 72 hours.

The mean score of observed values of edema & erythema calculated for each rabbit, after 24, 48, 72 hours, was equal to 0 for all the three rabbits.

 

Eye Irritation

The test substance was evaluated in the key study for its eye irritation potential according to the OECD guideline 405.

0.1 g of the test substance was instilled into the conjunctival sac of the left eye, of three rabbits. The eyelids were then held closed for some seconds. The right eye remained untreated and served as a control. The eyes of the animals were observed for 72 hours.

The mean scores of observed values of irritation/corrosion were calculated for each rabbit, after 24, 48, 72 hours.

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.

Justification for classification or non-classification

Skin Irritation/Corossion

A corrosive substance is a substance that produces destruction of skin tissue, namely, visible necrosis through the epidermis and into the dermis, in at least 1 tested animal after exposure up to 4 hour duration.

To classify a substance as irritant, the major criterion is that at least 2 of 3 tested animals have a mean score of ≥ 2,3 - ≤ 4,0.

The mean score of observed values of erythema & edema after 24, 48, 72 hours after patch removal for all the three rabbits is 0.

The substance is not classified as skin irritant because in the performed tests it doesn't meet the classification criteria of the CLP regulation (EC) No. 1272/2008.

 

Eye Irritation

Serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance to the anterior surface of the eye, which is not fully reversible within 21 days of application.

Eye irritation means the production of changes in the eye following the application of test substance to the anterior surface of the eye, which are fully reversible within 21 days of application.

The mean scores of observed values of irritation/corrosion were calculated for each rabbit, after 24, 48, 72 hours after installation of test material.

The corneal opacity in at least 2 of 3 tested animals was equal to 1 and the conjuctival redness was equal to 2 in all the three animals.

The test substance meets the requirements of the CLP regulation (EC) No. 1272/2008 , therefore it is classified in Category 2 (H19) for eye irritation.