Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-[(2-cyanoethyl)[2-(2-hydroxyethoxy)ethyl]amino]-1,3,5-triazine-4,2-diyl]imino]]bis[benzene-1,4-disulphonate]

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:

experimental study

Adequacy of study:

key study

Study period:

August from the 26th to the 30th, 1982

Reliability:

2 (reliable with restrictions)

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 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted on 12 May, 1981

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoering, AG.
- Age at start of treatment: ca. 12 weeks.
- Body weight at start of treatment: 2.3 - 3 kg.
- Diet: pelleted standard Kliba 23/341/1 rabbit maintenance diet, ad libitum.
- Water: tap water, ad libitum.
- Accomodation: individually in stainless steel cage.
- Acclimation period: 4 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Photoperiod: 12 hours light/dark.

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

other: polyethylene glycol (PEG 400) + saline (70 : 30 parts)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

1 g of a 50 % dilution of the test substance in polyethylene glycol (PEG 400) + saline (70 : 30 parts) was used.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

2 males and 1 females

Details on study design:

TEST SITE
- Area of exposure: 24 hours before the treatment, the back and flanks of each rabbit were closely clipped with electric clippers, exposing an area of skin of ca 10 x 10 cm. the test material was applied under a patch of surgical gauze approximately 3.0 x 3.0 cm.
- Type of wrap if used: the patch was covered with impermeable material and fastened to the body with adhesive tape.

OBSERVATION TIME POINTS
The skin reaction of the treated skin site was observed 1 hour, 24, 48, 72 hours, following removal of bandages and gauze patch.

SCORING SYSTEM
Draize 1959; Appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.

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 (raised approximately 1 mm) 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) 4

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:

Under the conditions of the test, the substance was found to cause no irritation when applied to intact rabbit skin.
In the area of application, no discolouration of the skin was observed in the rabbits which could be related to compound effects.
No corrosion effect had occurred on the skin at each measuring interval.

Animal	Reaction	1 hr	24 htrs	48 hrs	72 hrs
371 M	Erythema	0	0	0	0
372 M	Erythema	0	0	0	0
373 F	Erythema	0	0	0	0
371 M	Oedema	0	0	0	0
372 M	Oedema	0	0	0	0
373 F	Oedema	0	0	0	0

Interpretation of results:

other: not classified, according to CLP Regulation (EC) No1272/2008

Conclusions:

Not skin irritating

Executive summary:

The skin irritation potential of the test material was investigated by occlusive topical application to three New Zeland White rabbits. 1 g of a 50 % dilution of the test substance in polyethylene glycol (PEG 400) + saline (70 : 30 parts) was used.

The treated skin site was observed 1 hour, 24, 48, 72 hours, following removal of bandages and gauze patch and the skin reactions were scored in accordance with Draize system (1959) and Appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.

Under the conditions of the test, the substance was found to cause no irritation when applied to intact rabbit skin.

In the area of application, no discolouration of the skin was observed in the rabbits which could be related to compound effects.

No corrosion effect had occurred on the skin at each measuring interval.

Conclusion

The mean values from grading at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions.

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:

August from the 26th to the 30th, 1982

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 on 12 May, 1981

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoering, AG.
- Age at start of treatment: ca. 12 weeks.
- Body weight at start of treatment: 2.3 - 3 kg.
- Diet: pelleted standard Kliba 23/341/1 rabbit maintenance diet, ad libitum.
- Water: tap water, ad libitum.
- Accomodation: individually in stainless steel cage.
- Acclimation period: 4 days.
- Health check: before testing, both eyes of each animal were examined and any animal with eye defect was rejected.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Photoperiod: 12 hours light/dark.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g/rabbit

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

SCORING SYSTEM
The eye were examined and the grade of ocular reaction recorded at 1, 24, 48 and 72 hours after treatment.
Draize 1959; Appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.

Corneal opacity
No ulceration or opacity 0
Scattered or diffuse areas, details of iris clearly vsible 1
Easily discernible translucent area, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris not discernible through the opacity 4

Area of cornea affected
No ulceration or opacity 0
One quarter or less but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4

Iris
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia 1
No reaction to light, haemorrhage, gross destruction 2

Conjuctival redness 
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

Conjunctival oedema (chemosis)
No swelling 0
Any swelling above normal 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

Discarge
No discharge 0
Any amount different from normal 1
Discharge with moistening of the lids and hairs 2
Discharge with moistening of the lids and hairs, in a considerable area 3

In addition the colouration of sclera/cornea was recorded in connection with the colour of the test compound.

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Remarks on result:

no indication of irritation

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 in 2 out of 3 animals; no information about the 3rd animal because the test was stopped at 72 hrs

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:

Under the conditions of the test, the substance was found to cause no irritation when applied to rabbit eye mucosa.
No discolouration of the cornea and conjunctivae which could be related to compound effects was observed.
The primary irritaiton index was found to be 3.3
No corrosion effect was observed at each of the measuring intervals.

Individual reactions

Animal No	Reaction	1 hr	24 hrs	48 hrs	72 hrs	Mean 24/48/72 hrs
374 M	Cornea opacity	0	0	0	0	0.00
375 F	Cornea opacity	0	0	0	0	0.00
376 F	Cornea opacity	0	0	0	0	0.00
374 M	Iris	0	0	0	0	0.00
375 F	Iris	0	0	0	0	0.00
376 F	Iris	0	0	0	0	0.00
374 M	Conjunctival redness	2	1	1	1	1.00
375 F	Conjunctival redness	1	0	0	0	0.00
376 F	Conjunctival redness	2	1	1	0	0.67
374 M	Conjunctival chemosis	1	1	0	0	0.33
375 F	Conjunctival chemosis	1	0	0	0	0.00
376 F	Conjunctival chemosis	1	0	0	0	0.00
374 M	Conjunctival discharge	2	2	0	0	-
375 F	Conjunctival discharge	1	0	0	0
376 F	Conjunctival discharge	1	0	0	0

Interpretation of results:

other: not classified, according to CLP Regulation (EC) No1272/2008

Conclusions:

Not eye irritating

Executive summary:

The eye irritation potential of te test item was assessed in accordance with method and procedures outlined into the OECD guideline 405. 0.1 g of test substance were placed in the rabbit eyes, which were not rinsed. The eye were examined and the grade of ocular reaction recorded at 1, 24, 48 and 72 hours after treatment, according to the Draize system (1959) and in accordance with the Appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States criteria.

Under the conditions of the test, the substance was found to cause no irritation when applied to rabbit eye mucosa. No discolouration of the cornea and conjunctivae which could be related to compound effects was observed. The primary irritaiton index was found to be 3.3

No corrosion effect was observed at each of the measuring intervals.

Conclusion

The mean values from grading at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in all animals. No corrosion effect was observed at each of the measuring intervals. In one case the recovery of the redness cannot be judged because the test was stopped before; nevertheless, based on the severity of the effect observed and the decrease over the test period, the lacking of this information can be considered as not critical and not impacting the test outcomes conclusion.

Thus, the substance can be considered as non irritating to eye, 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

The skin irritation potential of the test material was investigated by occlusive topical application to three New Zeland White rabbits.1 g of a 50 % dilution of the test substance in polyethylene glycol (PEG 400) + saline (70 : 30 parts) was used. The treated skin site was observed 1 hour, 24, 48, 72 hours, following removal of bandages and gauze patch and the skin reactions were scored in accordance with Draize system (1959) and Appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States.

Under the conditions of the test, the substance was found to cause no irritation when applied to intact rabbit skin. In the area of application, no discolouration of the skin was observed in the rabbits which could be related to compound effects. No corrosion effect had occurred on the skin at each measuring interval.

In addition, the water-diluted substance was tested for skin irritation potential to rabbits. The irritation was measured by a patch test technique in 6 rabbits (males and females) and the resulting reactions were evaluated on the basis of the code of Federal Regulations, Title 16, Section 1500.41. In the intact skin neither erythema nor edema were observed. In the abraded skin after 24 hours, all six animals showed very slight or well defined erythema but no edema. After four days, all these findings had disappeared. In two aninals, the treated abraded area was slightly scaly.

Since the diluted substance was tested item, the results cannot be used for classification purposes; they have been included into the dossier exclusively for completeness sake.

EYE IRRITATION

The eye irritation potential of the test item was assessed in accordance with method and procedures outlined into the OECD guideline 405. 0.1 g of test substance were placed in the rabbit eyes, which were not rinsed. The eye were examined and the grade of ocular reaction recorded at 1, 24, 48 and 72 hours after treatment, according to the Draize system (1959) and in accordance with the Appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics, Association of Food and Drug Officials of the United States criteria.

Under the conditions of the test, the substance was found to cause no irritation when applied to rabbit eye mucosa. No discolouration of the cornea and conjunctivae which could be related to compound effects was observed. The primary irritation index was found to be 3.3

No corrosion effect was observed at each of the measuring intervals. In one case the recovery of the redness cannot be judged because the test was stopped before; nevertheless, based on the severity of the effect observed and the decrease over the test period, the lacking of this information can be considered as not critical and not impacting the test outcomes conclusion.

In addition, the water-diluted substance was tested for eye irritation potential to rabbits. 0.1 ml of the test material was placed into the conjunctival sac of the right eyes of the animals. Three males and three females were used; in addition, in three males treated eyes were washed after instillation; the left eye served as control. Reading score for ocular lesions were made according to the recommendations of the USA Food and Drug officials, published in 1959 (Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics).

No ocular findings at all were observed neither in the unrinsed nor the rinsed eye.

Since the diluted substance was tested item, the results cannot be used for classification purposes; they have been included into the dossier exclusively 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.

The mean values from grading at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions.

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 grading 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 animals.

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