Disodium ar,ar'-[(9,10-dihydro-9,10-dioxo-1,4-anthrylene)diimino]bis[(4-chlorophenoxy)benzenesulphonate]

Endpoint summary

Description of key information

Not classified as skin irritating, according to the CLP Regulation (EC) No 1272/2008

Eye Irrit. 2 (H319), according to the CLP Regulation No (EC) No 1272/2008

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

The procedure employed was that described by the Consumer Product Safety Commission of the U.S.A. in The Code of Federal Regulations, Title 16, Section 1500.41.

GLP compliance:

no

Remarks:

pre GLP

Species:

rabbit

Type of coverage:

occlusive

Preparation of test site:

other: abraded and intact skin

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 g of test item were mixed with 0.5 ml of distilled water

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

six rabbits

Details on study design:

TEST SITE
- Area of exposure: two slngle layers thick moinstoned with test item were introduce under a square patch such as surgical gauze measuring 1 inch x 1 inch.
- Type of wrap if used: the entire trunk of the animal was wrapped with an impervious material such as rubberized cloth.

SCORING SYSTEM
Readings are again made at the end of a total of 72 hours (48 hours after the first reading). An equal number of exposures are made on areas of skin that have been previously abraded. The abrasions are minor incisions through the stratum corneum, but not sufficiently deep to disturb the derma or to produce bleeding. Evaluate the reactions of the abraded skinat 24 hours and 72 hours. Add the values for erythema and eschar formation at 24 hours and at 72 hours for intact skin to the values on abraded skin at 24 hours and at 72 hours (four values). Slmilarly, add the values for oedema formation at 24 hours and at 72 hours for intact and abraded skln (four values). The total of the eight values is divlded by four to give the primary irritation score.

Erythema and eschar formation
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Oedema formation
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema ( ratsed approximately I milllmeter) 3
Severe oedema (raised more than 1 miillimeter and extending beyond the area of exposure) 4

Classification system recommended by the ETAD subcommittee for Toxicology:
0 - 0.5 non-irritant
0.6 - 3.0 slightly irritant
3.1 - 5.0 moderately irritant
5.1 - 8.0 severely irritant

Irritation parameter:

erythema score

Basis:

animal: 6/6

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Remarks:

none of the animals showed any observable response

Irritation parameter:

edema score

Basis:

animal: 6/6

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Remarks:

none of the animals showed any observable response

Irritant / corrosive response data:

None of the animals showed any observable response to treatment throughout the 72 hours observation period.

Interpretation of results:

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

Conclusions:

Not irritating

Executive summary:

Primary irritation to the skin was assayed following the procedures prescribed by The Consumer Product Safety Commission of the U.S.A. in The Code of Federal Regulations, Title 16, Section 1500.41. 0.5 g of test item were rnixed with 0.5 ml of distilled water before application to the skin. The primary irritation was measured by a patch technique on both abraded and intact skin of albino rabbits.

None of the animals showed any observable response to treatment throughout the 72 hours observation period.

Conclusion

The scoring system criteria used to record the hardness of the skin reactions followed in the current test are the same as those reported into the OECD guideline, therefore it is possible to re-evaluate the results in accordance with the CLP Regulation (EC) No 1272/2008.

None of the animals showed any observable response to treatment throughout the 72 hours observation period, thus it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 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:

March from 09 to 20, 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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 CH 4414 FueIllinsdorf/SwitzerIand.
- Age at study initiation: 14 - 15 weeks.
- Weight at study initiation: 2.4 - 2.8 kg.
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system.
- Diet: pelleted standard KIiba 341, Batch 94/84 rabbit maintenance diet, ad Iibitum.
- Water: community tap water from Itingen, 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 hours artificial fluorescent light/12 hours dark.
- Other: at least 8 hours music/light period.

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

2 males and 1 female

Details on study design:

TOOL USED TO ASSESS SCORE: eye examinations were made with a slit-lamp 30 SL.

OBSERVATIONS
- Viability / Mortality: daily.
- Body weights: pre-test, day 1 and at termination.

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 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.
The following subjective numerical scoring system was used to grade the eye lesions.

CORNEAL IRRITATION
Opacity, degree of density (densest area used for assessment)
No ulceration of opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacreous area for details of iris visible, size of pupil banely discernible 3
Opaque cornea, iris not discernible through the opacity 4

IRIDIC IRRITATION
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circurncorneal hyperemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, hemorrahge, gross destruction (any or all of these) 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrar and bulbar conjunctivae, cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperaemic (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 (include nictating membranes) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

>= 1 - <= 1.7

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 48 hrs

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

>= 2.3 - <= 3

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:

Test item showed a primary irritation score of 5.0 when applied to the rabbit eye mucosa.
In the area of application a bluish discoloration of the cornea and conjunctivae was observed within 1 hour after treatment. This could be related to effects of the test article.
No corrosion of the cornea was observed at each of the measuring intervals.

Other effects:

TOXIC SYMPTONS / 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.

MEAN SCORES PER ANIMAL

Animal N. / sex	Reaction	1 hr	24 hrs	48 hrs	72 hrs	7 days	Mean 24/48/72 hrs
323 M	Corneal opacity	1*	1*	1*	1*	0	1.00
324 M	Corneal opacity	2*	2*	1*	1*	0	1.33
325 F	Corneal opacity	1*	2*	2*	1*	0	1.66
323 M	Iris	0	1*	0	0	0	0.33
324 M	Iris	0	1*	0	0	0	0.33
325 F	Iris	0	1*	0	0	0	0.33
323 M	Conjunctival redness	1	3*	3*	3*	1	3.00
324 M	Conjunctival redness	1	3*	3*	2*	0	2.67
325 F	Conjunctival redness	1	3*	2*	2*	0	2.33
323 M	Conjunctival chemosis	2*	2*	1	0	0	1.00
324 M	Conjunctival chemosis	3*	1	1	0	0	0.67
325 F	Conjunctival chemosis	2*	1	1	0	0	0.67

INDIVIDUAL FINDINGS

Animal No 323, male

After 1 hr	Cornea: opacity, slight, whole; discolored. Conjunctivae: reddened, slight, dorsal; edema, clear visible; discolored, including nictating membrane, substance remainders, greenish mucous diffuse. Sclera: not visible. Discharge: severe.
After 24 hrs	Cornea: opacity, whole, diffuse. Iris: red. Conjunctivae: reddened severe; edema. Sclera: reddened, severe. Discharge: moderate to severe.
After 48 hrs	Cornea: opacity ventral. Conjunctivae: reddened severe; edema slight. Sclera: reddened, severe. Discharge: minimal
After 72 hrs	Cornea: opacity ventral, diffuse, hardly visible. Conjunctivae: reddened severe. Sclera: reddened, severe.
After 7 days	Conjunctivae: reddened, conjunctival sac, hardly visible.

Animal No 324, male

After 1 hr	Iris: darkened. Conjunctivae: reddened, slight; edema, severe, discolored including nictating membrane; substance remainders, greenish mucous, diffuse. Sclera: not visible. Discharge: severe.
After 24 hrs	Cornea: opacity, whole. Iris: red. Conjunctivae: reddened, severe, whole; edema, slight. Sclera: reddened, severe. Discharge: severe.
After 48 hrs	Cornea: opacity, whole. Conjunctivae: reddened, severe, whole; edema, slight. Sclera: reddened, severe.
After 72 hrs	Cornea: opacity, whole. Conjunctivae: reddened, clear visible. Sclera: reddened, ventral
After 7 days	No abnormal changes visible.

AnimaI No 325, female

After 1 hr	Cornea: opacity, slight, whole; discolored. Conjunctivae: reddened, slight, dorsal; edema, clear visible; discolored, including nictating mernbrane, substance remainders, greenish rnucous, diffuse. Sclera: not visible. Discharge: severe.
After 24 hrs	Cornea: opacity, whole. Iris: red, slight. Conjunctivae: reddened, severe; edema, slight. Sclera: reddened, slight. Discharge: moderate.
After 48 hrs	Cornea: opacity, diffuse. Conjunctivae: reddened, clear visible; edema slight. Sclera: reddened, clear visible. Discharge: minimal.
After 72 hrs	Cornea: opacity, ventral slight. Conjunctivae: reddened, clear visible. Sclera: reddened, clear visible. Discharge: moderate.
After 7 days	No abnormal changes visible.

Interpretation of results:

other: Eye Irrit. 2 (H319), according to the CLP Regulation No (EC) 1272/2008

Conclusions:

Test item is able to cause eye irritation.

Executive summary:

The primary eye irritation study was investigate according to the OECD guideline 405. The irritation potential was assessed placing the test item in the conjunctival sac of rabbit eyes. In the area of application a greenish discoloration of the cornea and conjunctivae was observed within 1 hour after treatment. This could be related to effects of the test article. The mean values from gradings at 24, 48 and 72 hours were between 1 and 1.7 for corneal opacity, lower than 1 for iritis, between 2.3 and 3 for conjunctival redness and lower than 2 for conjunctival oedema, in all of the tested animals. No corrosion was observed at each of the measuring intervals.

The substance was found to cause a primary irritation score of 5.0 when applied to the rabbit eye mucosa.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were between 1 - 1.7 for corneal opacity, lower than 1 for iritis, between 2.3 - 3 for both conjunctival redness and lower than 2 for conjunctival oedema, in all of the tested animals. All the reactions were fully recovered within 7 days.

The test item meets the criteria to be classified as capable to causes serious eye irritation (i.e. Eye Irrit. 2, H319), according to the CLP Regulation No (EC 1272/2008).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

SKIN IRRITATION

Primary irritation to the skin was assayed following the procedures prescribed by The Consumer Product Safety Commission of the U.S.A. in The Code of Federal Regulations, Title 16, Section 1500.41. The primary irritation was measured by a patch technique on both abraded and intact skin of albino rabbits. None of the animals showed any observable response to treatment throughout the 72 hours observation period.

EYE IRRITATION

The primary eye irritation study was investigate according to the OECD guideline 405. The irritation potential was assessed placing 0.1 g of the test item in the conjunctival sac of rabbit eyes. In the area of application a greenish discoloration of the cornea and conjunctivae was observed within 1 hour after treatment. This could be related to effects of the test article. The mean values from gradings at 24, 48 and 72 hours were between 1 and 1.7 for corneal opacity, lower than 1 for iritis, between 2.3 and 3 for conjunctival redness and lower than 2 for conjunctival oedema, in all of the tested animals. No corrosion was observed at each of the measuring intervals. The substance was found to cause a primary irritation score of 5.0 when applied to the rabbit eye mucosa.

A second old experiment is also available. The test item eye irritation potential was assayed following the procedures prescribed by The Consumer Product Safety Commission of the U.S.A. in The Code of Federal Regulations, Title 16, Section 1500.42. Corneal opacities developed in three animals. Iritis was observed in two animals. A diffuse crimson red colouration of the conjunctivae with considerable swelling and partial eversion of the eyelids or with the eyelids about half closed was observed in two animals. Mild conjunctival reactions were observed in three animals. One animal did not show any observable response to treatment throughout the 7 days observation period. 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 the tested animals. In this case, the substance caused less sever effect; this can be explained by the composition of the lot tested, in which the test item content was lower than the content in the lot tested in the key study.

An additional experiment was conducted with Acid Green 040; however, the complete study report is not more available and only a sheet reporting the conclusive classification can be retrieved. The substance was classified as moderately irritant, according to the directive of the VCI (Verband der Chemischen Industrie e.V., FRG) and of the ETAD (Ecological and Toxicological Association of the Dyestuff Manufacturing Industries), Methods 001 - 003 and it was classified as irritant according to the criteria described in the Commission Directive (of adapting to technical progress for the fifth time Council Directive 67/548/EEC, Annex VI, Part IIB).

Due to the fact that details on testing method, procedures and results are completely lacking, a reliability cannot be assigned; data is here mentioned only for completeness sake.

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.

None of the animals showed any observable response to treatment throughout the 72 hours observation period, thus it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.

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. 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 between 1 - 1.7 for corneal opacity, lower than 1 for iritis, between 2.3 - 3 for both conjunctival redness and lower than 2 for conjunctival oedema, in all of the tested animals. All the reactions were fully recovered within 7 days.

In conclusion, the substance is not classified for the skin irritation; however, the substance meets the criteria to be classified as eye irritating, category 2 (H319), according to the CLP Regulation (EC) No. 1272/2008.