Sodium 1-amino-9,10-dihydro-9,10-dioxo-4-[(2,2,4-trimethylcyclohexyl)amino]anthracene-2-sulphonate

Endpoint summary

Description of key information

Not skin irritating

Not eye irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From May 26th to August 15th, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The read across approach is detailed into the document attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

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: the rabbits were house 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 × 6 cm was depilated. For the formation of scarified skin, four horizontal and four vertical incisions were made. Depilated area of​​ skin outside of the application zone was used to control the assessment.
- 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.

SCORING SYSTEM
The degree of erythema was diagnosed visually, and the degree of edema was assessed by visual examination and palpation, accordign to the OECD guideline 404.

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

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.

Animal No	Reaction	Intact skin				Scarified skin
		4 hrs	24 hrs	48 hrs	72 hrs	4 hrs	24 hrs	48 hrs	72 hrs
14	Erythema	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0
29	Erythema	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0
30	Erythema	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0

Interpretation of results:

other: not classified, according to the CLP Regulation (EC1272/2008)

Conclusions:

Not irritant.

Executive summary:

The substance was evaluated for skin irritation/ corossion, according to OECD guideline 404. The test substance (0.5 g) 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 and 72 hours of the removal of the patch the animals were examined for signs of erythema and ethema. 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.

Conclusion

None of the animals showed any observable response to treatment throughout the 72 hours observation period, in both the cases of intact and scarified skin. Therefore, it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC 1272/2008).

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 February 24th to March 06th, 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted May 12, 1981

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, Fuellinsdorf / Switzerland.
- Age at study initiation: 14 - 15 weeks.
- Weight at study initiation: 2.4 - 2.5 kg.
- Housing: individually in steinless steel cage, equipped with automatic cleaning and drinking system.
- Diet: pelleted standard Kilba 341, batch 93/83, rabbit maintenance diet, ad libitum.
- Water: community tap water, ad libitum.
- Acclimation period: 4 days under laboratory conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Relative humidity: 55 ± 10 %
- Air changes: 10 - 15 air change per hour.
- Photoperiod: 12 hrs artificial light / hrs darkness.
- Other: at least 8 hours of music / light period.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g per animal

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

1 maes and 2 females

Details on study design:

TOOL USED TO ASSESS SCORE: hand-slit lamp 30 SL and a Varta Cliptrix diagnostic-lamp.

OBSERVATIONS
- Viability/mortality: daily
- Body weight: pre-treatment, day 1 and at termination of the test.

SCORING SYSTEM
The rabbits were examined 1, 24, 48, 72 hours and 7 days after application of the test compound. The ocular reactions were scored according to the OECD guideline 405.

Opacity-degree of density (area most dense taken for reading)
No opacity 0
Scattered or diffuse area (other than slight dulling of normal luster) details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Nacros area, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4

Iris
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any 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

Conjunctivae Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperemic (injected) 1
Diffuse, crimson color, individual vessels not easily discernible 2
Diffuse beefy red 3

Chemosis: lids and/or nictating membranes
No swelling 0
Any swelling above normal (includes nictitating membranes) 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

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 72 hrs

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 48 hrs

Irritant / corrosive response data:

Test substance showed a primary irritation score of 2.1 when applied to the rabbits eye mucosa.
In the area of application a bluish discoloration of the cornea, sclera and conjunctivae was observed within 1 hour after treatrnent. This could be related to effects of the test article
No corrosion of the cornea was observed at each of the measuring intervals..

TOXIC SYMPTOMS / MORTALITY
No acute toxic symptoms were observed in the animals during the test period and no mortality occurred.

BODY WEIGHT
The body weight gain of all rabbits was similar.

Eye irritation scores

Animal N / sex	Reaction	1 hr	24 hrs	48 hrs	72 hrs	7 days	Mean 24/48/72 hrs
281 M	Cornea opacity	0	1*	1*	0	0	0.67
282 F	Cornea opacity	0	1*	1*	0	0	0.67
283 F	Cornea opacity	1*	1*	1*	0	0	0.67
281 M	Iris	0	0	0	0	0	0.00
282 F	Iris	0	0	0	0	0	0.00
283 F	Iris	0	0	0	0	0	0.00
281 M	Conjunctivae redness	0	1	1	1	0	1.00
282 F	Conjunctivae redness	0	1	1	1	0	1.00
283 F	Conjunctivae redness	0	1	1	1	0	1.00
281 M	Conjunctivae chemosis	2*	1	0	0	0	0.33
282 F	Conjunctivae chemosis	2*	2*	0	0	0	0.67
283 F	Conjunctivae chemosis	2*	1	0	0	0	0.33

Primary irritation score

Time point	Mean scores
1 hr	2.3
24 hrs	3.3
48 hrs	2.0
72 hrs	1.0

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Not irritating

Executive summary:

A study was performed to assess the irritation of the test material to the eye of the New Zealand White rabbit. The method followed OECD Guidelines No. 405. A single application 0.1 g of the test material was administered to the eye of three rabbits. Ocular effects were recorded 1, 24, 48 and 72 hours and 7 days after administration.

Test substance showed a primary irritation score of 2.1 when applied to the rabbits eye mucosa. In the area of application a bluish discoloration of the cornea, sclera and conjunctivae was observed within 1 hour after treatrnent. This could be related to effects of the test article. No corrosion of the cornea was observed at each of the measuring intervals.

No acute toxic symptoms were observed in the animals during the test period and no mortality occurred. The body weight gain of all rabbits was similar.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all of three tested animals. Therefore, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC) No 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

SKIN IRRITATION

No information on the skin irritation/corrosion potential of the Target Substance is available. For this reason data coming from an analogue substance (Similar Substance 01) has been taken into account. More details aboit the similarity between the two substances is reported in the document attached in section 13.

The substance was evaluated for skin irritation/ corrosion, according to OECD guideline 404. The test substance (0.5 g) 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. After 4, 24, 48 and 72 hours of the removal of the patch the animals were examined for signs of erythema and ethema; 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.

EYE IRRITATION

A study was performed to assess the eye irritation potential of Acid Blue 278 to New Zealand White rabbit. The method followed OECD Guidelines No. 405. A single application 0.1 g of the test material was administered to the eye of three rabbits. Th eyes were not rinsed. Test substance showed a primary irritation score of 2.1 when applied to the rabbits eye mucosa. In the area of application a bluish discoloration of the cornea, sclera and conjunctivae was observed within 1 hour after treatrnent. No corrosion of the cornea was observed at each of the measuring intervals. No acute toxic symptoms were observed in the animals during the test period and no mortality occurred. The body weight gain of all rabbits was similar.

In addition, a sheet reporting the test results of an old experiment conducted on Acid Blue 278 is available. The effects were scored in accordance to the ETAD system. However, due to the fact that details on testing procedures and results are lacking, a reliability cannot be assiged. The result is here mentioned for completeness sake. The substance was indicated to be slightly irritating (Sheet of test results, 1977).

Justification for classification or non-classification

According to the CLP Regulation (EC) No 1272/2008, 3.2 Skin corrosion/irritation section, skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

On the basis of the available information, the substance was not classified.

According to the CLP Regulation (EC) No 1272/2008, serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance, which is not fully reversible within 21 days of application.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all the tested animals. In addition, all the recorded effects were fully reversible within 72 hours.

In conclusion, the substance does not meet the criteria to be classified for the eye/skin irritation, according to the CLP Regulation (EC) No 1272/2008.