Sodium 2-[[4-(cyclohexylamino)-9,10-dihydro-9,10-dioxo-1-anthryl]amino]-5-ethoxybenzenesulphonate

Endpoint summary

Description of key information

Non skin irritating

Non 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

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

May 12, 1981

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH 4414 Fuellinsdorf / Switzerland.
- Age at study initiation: 14 - 15 weeks
- Weight at study initiation: 2.4 - 2.9 kg.
- Housing: individually in stainless steel cages equipped with a automatic cleaning and drinking system.
- Diet: pelleted standand KIiba 341, Batch 94/84 rabbit maintenance diet ("KIiba", Klingentalmuehle AG, Switzerland), ad libitum.
- Water: community tap water from Itigen, ad libitum.
- Acclimation period: four days under Iaboratory conditions, after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Air changes: 15 air changes per hr.
- Photoperiod: 12 hrs dark / 12 hrs light.
- Other: at least 8 hours music/light period.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A single dose was administrated to the left eye of each animal. The individual dose volume was 0.1 g per animal.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: eyes were not rinsed.

OBSERVATIONS
- Viability/mortality: daily.
- Body weight: pre-test, day 1 and at termination of test on Mettler PK 16 balance.

SCORING SYSTEM
The eyes of each animal were examined 1, 24, 48, 72 hours and 7 days after administration. The irritation was assessed according to the OECD guideline for testing of Chemicals, Section 4, number 405 "Acute Eye Irritation/Corrosion" adopted May 12, 1981.
The corrosive properties of the test article and the color of the treated eye were described and recorded.

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 48 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: 72 hrs

Irritant / corrosive response data:

Under the conditions of the experiment, the test item was found to cause a primary irritation score of 2.2 when applied to the rabbit eye mucosa.
In the area of application a bluish dicoloration of the conjunctivae was observed within 1 hour after treatment; this could be related to effects of the test article.
No corrosion was observed at each of the measuring intervals.

Other effects:

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

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

Eye irritation score

Animal N.	Reaction	After					Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs	7 days
320 M	Cornea opacity	1	1	0	0	0	0.33
321 F	Cornea opacity	0	0	0	0	0	0.00
322 F	Cornea opacity	0	0	0	0	0	0.00
320 M	Iris	0	0	0	0	0	0.00
321 F	Iris	0	0	0	0	0	0.00
322 F	Iris	0	0	0	0	0	0.00
320 M	Conjuntivae redness	1	2	2	1	0	1.67
321 F	Conjuntivae redness	1	1	1	1	0	1.00
322 F	Conjuntivae redness	1	1	1	1	0	1.00
320 M	Conjunctivae chemosis	2	2	1	0	0	1.00
321 F	Conjunctivae chemosis	2	1	0	0	0	0.33
322 F	Conjunctivae chemosis	2	1	0	0	0	0.33

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

Not irritating

Executive summary:

Three rabbits were used to test the eye irritation potential of the substance, in accordance with OECD guideline 405. 0.1 g of the undiluted test material were placed into the conjunctival sac. The eyes of each animal were examined 1, 24, 48, 72 hours and 7 days after administration. Reading scores for ocular lesions were made according to the OECD guideline for testing of Chemicals, Section 4, number 405 "Acute Eye Irritation/Corrosion" adopted May 12, 1981.

Under the conditions of the experiment, the test item was found to cause a primary irritation score of 2.2 when applied to the rabbit eye mucosa. In the area of application a bluish dicoloration of the conjunctivae was observed within 1 hour after treatment; this could be related to effects of the test article. No corrosion was observed at each of the measuring intervals.

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 chemosis, in all the tested animals. All the reactions were fully reversible.

Thus, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC 1272/2008).

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Study period:

June from 01st to 04th, 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted February 24, 1987

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH 4414 Fuellinsdorf / Switzerland.
- Age at study initiation: 14 - 15 weeks
- Weight at study initiation: 2.8 kg.
- Housing: individually in stainless steel cages equipped with a automatic cleaning and drinking system.
- Diet: pelleted standand KIiba 341, Batch 25/87 and 26/87 rabbit maintenance diet ("KIiba", Klingentalmuehle AG, Switzerland), ad libitum.
- Water: community tap water from Itigen, ad libitum.
- Acclimation period: five days under Iaboratory conditions, after veterinary examination. The eyes of the animals were observed 24 hours prior to test article administration.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3 °C
- Humidity: 40 - 70 %
- Air changes: 10 - 15 air changes per hr.
- Photoperiod: 12 hrs dark / 12 hrs light.
- Other: music/light period.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A single dose was administrated to the left eye of each animal. The individual dose volume was 0.1 g per animal.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

2 males and 1 female

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: eyes were not rinsed.

OBSERVATIONS
- Viability/mortality: daily.
- Body weight: pre-test, day 1 and at termination of test on Mettler PK 16 balance.

SCORING SYSTEM
The eyes of each animal were examined 1, 24, 48, 72 hours and 7 days after administration. The irritation was assessed according to the OECD guideline for testing of Chemicals, Section 4, number 405 "Acute Eye Irritation/Corrosion" adopted May 12, 1981.
The corrosive properties of the test article and the color of the treated eye were described and recorded.

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: 72 hrs

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 72 hrs

Irritant / corrosive response data:

Under the conditions of the experiment, the test item was found to cause a primary irritation score of 1.9 when applied to the rabbit eye mucosa.
Corneal opacity was observed in two animals duning the test. Anlmal no. 458 (male) showed sllght opacity from 1 to 48 hours, anirnal no. 459 (female) showed diffuse opacity from 1 to 24 hours after test article application.

Other effects:

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

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

Eye irritation score

Animal N.	Reaction	After				Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs
457 M	Cornea opacity	0	0	0	0	0.00
458 M	Cornea opacity	1	1	1	0	0.67
459 F	Cornea opacity	1	1	0	0	0.33
457 M	Iris	0	0	0	0	0.00
458 M	Iris	0	0	0	0	0.00
459 F	Iris	0	0	0	0	0.00
457 M	Conjuntivae redness	1	1	1	0	0.67
458 M	Conjuntivae redness	1	1	1	0	0.67
459 F	Conjuntivae redness	1	1	1	0	0.67
457 M	Conjunctivae chemosis	1	0	0	0	0.00
458 M	Conjunctivae chemosis	2	2	1	0	1.00
459 F	Conjunctivae chemosis	1	1	0	0	0.33

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

Not irritating

Executive summary:

Three rabbits were used to test the eye irritation potential of the substance, in accordance with OECD guideline 405. 0.1 g of the undiluted test material were placed into the conjunctival sac. The eyes of each animal were examined 1, 24, 48, 72 hours and 7 days after administration. Reading scores for ocular lesions were made according to the OECD guideline for testing of Chemicals, Section 4, number 405 "Acute Eye Irritation/Corrosion" adopted May 12, 1981.

Under the conditions of the experiment, the test item was found to cause a primary irritation score of 1.9 when applied to the rabbit eye mucosa.

Corneal opacity was observed in two animals duning the test. Anlmal no. 458 (male) showed sllght opacity from 1 to 48 hours, anirnal no. 459 (female) showed diffuse opacity from 1 to 24 hours after test article application.

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 chemosis, in all the tested animals. All the reactions were fully reversible within 72 hours.

Thus, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC 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

An old experiment was conducted on the test item; however, due to the fact that details on testing procedures and results are lacking, a reliability cannot be assigned. The result is here mentioned for completeness sake. The skin irritation potential was assayed in rabbits and scored according to the U.S. Hazardous Substance Act of 16.09.1964.. The substance was indicated to be non skin irritating (see attachment).

In order to assess the skin irritation potential of Acid Blue 280, the available information on the structural analogue Similar Substance 03 was taken into consideration; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID 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

An old experiment was conducted on the test item; however, due to the fact that details on testing procedures and results are lacking, a reliability cannot be assigned. The result is here mentioned for completeness sake. The eye irritation potential was assayed in rabbits and scored according to the U.S. Hazardous Substance Act of 16.09.1964.. The substance was indicated to be non eye irritating (see attachment).

In order to clarify the eye irritation potential of Acid Blue 280, the available information on the structural analogues Similar Substance 02 was taken into consideration; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID section 13).

Two different experiments are available; in both the cases, Similar Substance 02 was tested in accordance with OECD guideline 405 and 0.1 g of the undiluted test material were placed into the conjunctival sac.

Under the conditions of the first experiment, the test item was found to cause a primary irritation score of 1.9 when applied to the rabbit eye mucosa. Corneal opacity was observed in two animals duning the test. Animal no. 458 (male) showed sllght opacity from 1 to 48 hours, anirnal no. 459 (female) showed diffuse opacity from 1 to 24 hours after test article application.

In the second experiment, the test item was found to cause a primary irritation score of 2.2 when applied to the rabbit eye mucosa. In the area of application a bluish dicoloration of the conjunctivae was observed within 1 hour after treatment; this could be related to effects of the test article. No corrosion was observed at each of the measuring intervals.

In both the experiments, 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 chemosis, in all the tested animals. All the reactions were fully reversible within 72 hours.

REFERENCE

See attachment

Justification for classification or non-classification

According to the CLP Regulation (EC 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.

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.

According to the CLP Regulation (EC 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. Eye irritation means the production of changes in the eye, which are 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 chemosis, in all the tested animals. All the reactions were fully reversible within 72 hours.

In conclusion, the substance is not classified for the eye/skin irritation, according to the CLP Regulation (EC 1272/2008).