tetrasodium 2-(4-fluoro-6-(methyl-(2-(sulfatoethylsulfonyl)ethyl)amino)-1,3,5-triazin-2-ylamino)-5-hydroxy-6-(4-methyl-2-sulfonatophenylazo)naphthalene-1,7-disulfonate

Endpoint summary

Description of key information

The test substance is non-irritating to the skin or eye of rabbits. It caused discolouration of conjunctivae, nictitating membranes and sclera which were not completely reversed at the end of the observation period.

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 August 17, 1999 to August 20, 1999

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

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: 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: Charles River Deutschland GmbH 88353 Kißlegg.
- Body weight range at treatment: 2.8-3.4 kg
- Housing: In fully air-conditioned rooms in separate cages arranged in a battery
- Diet: Ssniff K-H (V2333), ad libitum and hay (i.e., approximately 15 g daily)
- Water: Water from automatic water dispensers, ad libitum
- Acclimation period: 1 week under study conditions
- Animal identification: Numbered ear tags

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Humidity: 50±20%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: August 17, 1999 to August 20, 1999

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

500 mg (i.e., 0.5g)

Duration of treatment / exposure:

4 h

Observation period:

30 - 60 minutes as well as 24, 48 and 72 h after removal of the patches.

Number of animals:

3 females

Details on study design:

TEST PROCEDURE
About 24 h before the start of the study the hair in the dorsal region of the body of 3 rabbits was removed with an electric clipper over an area of about 25 cm2. Only animals with intact skin were used.

Each animal was treated with 0.5 g test substance pasted with 0.5 mL deionized water. The substance was administered over the whole surface of a 2.5 x 2.5 cm cellulose patch on a piece of surgical plaster (i.e., specially produced by Beiersdorf Aktiengesellschaft). The plaster was fixed to the prepared skin area and then covered with a semi-occlusive bandage.

The exposure period was 4 h. After the exposure period all remnants of the test substance were carefully removed from the skin with warm tap water.

Examinations of the skin took place after 30 - 60 minutes as well as 24, 48 and 72 h after removal of the patches.

Erythema, eschar formation and edema were evaluated numerically according to the score of Draize . All other changes of the skin were recorded.

SCORING SYSTEM:

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 eschar formation (injuries in depth) preventing
grading of erythema...........................................................................................................4
Not assessable.....................................................................................................................*
Edema
No edema..............................................................................................................................0
Very slight edema (barely perceptible).............................................................................1
Slight edema (edges of area well defined by definite raising).......................................2
Moderate edema (raised approximately 1 mm) ..............................................................3
Severe edema (raised more than 1 mm and extending beyond area of exposure)...4
Not assessable......................................................................................................................*

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Other effects:

30 - 60 min up to 1 d after removal of the plaster the animals showed orange discolored skin.
3 d after administration all discolorations were disappeared.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test substance was considered to be non-irritating to skin.

Executive summary:

A study was performed to assess the skin irritation potential of the test substance in femaleNew Zealand albino rabbit according to OECD Guideline 404 and EU Method B.4, in compliance with GLP.

500 mg of test substance pasted with 0.5 mL deionized water was evenly distributed over the whole surface of a 2.5 x 2.5 cm cellulose patch on a piece of surgical plaster. The plaster was fixed to the prepared skin area of about 25 cm² and then covered with a semi-occlusive bandage. The patch was allowed to remain in contact with the skin for 4 h. After the exposure period, all remnants of test substance were carefully removed with warm tap water. Animals were scored for irritation reactions after 30 - 60 min, and at 24, 48 and 72 h.

Except for orange discolored skin in treated animal up to 1 d after removal of the plaster, no severe signs of irritation were observed during the whole observation period. Further, all discolorations were reversiblewithin 3 d. Overall mean scores for dermal irritation were calculated to be 0.0 for erythema and eschar formation and 0.00 for edema formation.

Under the test conditions, the test substance was considered to be non-irritating to 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 August 24, 1999 to September 14, 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

only one animal was tested. As the persistent discolouration of the conjunctivae requested a classification according to EU Directive 67/548/EEC, no further animals were tested.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: Charles River Deutschland GmbH, 88353 Kisslegg.
- Body weight at treatment: 3.0 kg
- Housing: In fully air-conditioned rooms in separate cages arranged in a battery
- Diet: Ssniff K-H (V2333), ad libitum and hay (i.e., approximately 15 g daily)
- Water: Water from automatic water dispensers, ad libitum
- Acclimation period: 1 week under study conditions
- Animal identification: Numbered ear tags

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Humidity: 50±20%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: August 24, 1999 to September 14, 1999

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48 , 72 h and after 7, 14 d and 21 d following administration of the test substance.

Number of animals or in vitro replicates:

1 female

Details on study design:

TEST PROCEDURE

About 24 h before the start of the study the test eyes of all animals were examined under UV light for corneal lesions after instillation of one drop of a 0.01% fluorescein-sodium solution. Only animals without ocular abnormalities were used for the study.

100 mg test substance was administered once to the conjunctival sac of the left eye of one rabbit. In each case the untreated eye served as a control. 24 h after administration the treated eye was washed out thoroughly with isotonic saline at approximately 37ºC. The eye was also washed out at designated examination times at which discharge was observed or a corneal examination with fluorescein was performed.

The eye was examined 1, 24, 48 and 72 h after administration of the test substance. At 24 and 72 h and after 7, 14 and 21 d, the eye was further examined for corneal lesions under UV light after instillation of one drop of a 0.01% fluorescein-sodium solution. Lesions in cornea, iris or conjunctivae were graded numerically. All other changes or toxic effects were recorded when apparent. Since effects were still present in the eye after 72 h, further examinations were carried out after 7, 14 and 21 d.

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 2 d

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

- From 1 h up to 1 d after application the conjunctiva of the animal showed definitely injected blood vessels up to a diffuse crimson red color and swelling with the lid being half closed.
- The irritations were attended by clear substance coloured eye discharge.
- 2 d after application the irritations were reversed.
- Additionally, nictitating membrane, conjunctiva and sclera were discolored orange from 1 h up to the end of the study.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the substance caused orange discoloration of the conjunctiva, nictitating membrane and sclera of one rabbit eye until end of the study period. Reddening of the conjunctiva was seen after substance application but this cleared within two days.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in one New Zealand albino female rabbit according to OECD Guideline 405 and EU Method B.5, in compliance with GLP.

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of the animal. The treated eye was washed out 24 h after administration. The untreated eye served as a control.

The conjunctiva, nictitating membrane and sclera were discolored orange by the test substance up to the end of the study. The conjunctiva showed definitely injected blood vessels up to a diffuse crimson red color and swelling with the lid being half closed, accompanied by clear substance coloured eye discharge on the day of substance administration. Based on the individual scores after 24, 48 and 72 h, the mean score was 0 for opacity of the cornea, iris and chemosis of the conjunctiva. The score was 0.33 for redness of the conjunctiva.

Under the test conditions, the substance caused orange discoloration of the conjunctiva, nictitating membrane and sclera of one rabbit eye until end of the study period. Reddening of the conjunctiva was seen after substance application but this cleared within two days.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

A study was performed to assess the skin irritation potential of the test substance in female New Zealand albino rabbit according to OECD Guideline 404 and EU Method B.4, in compliance with GLP. 500 mg of test substance pasted with 0.5 mL deionized water was evenly distributed over the whole surface of a 2.5 x 2.5 cm cellulose patch on a piece of surgical plaster. The plaster was fixed to the prepared skin area of about 25 cm² and then covered with a semi-occlusive bandage. The patch was allowed to remain in contact with the skin for 4 h. After the exposure period, all remnants of test substance were carefully removed with warm tap water. Animals were scored for irritation reactions after 30 - 60 min, and at 24, 48 and 72 h. Except for orange discolored skin up to 1 d after removal of the plaster, no severe signs of irritation were observed in treated animals during the whole observation period. Further, all discolorations were reversible within 3 d. Overall mean scores for dermal irritation were calculated to be 0.0 for erythema and eschar formation and 0.00 for edema formation. Under the test conditions, the test substance was considered to be non-irritating to skin (Seeberger, 1999b).

Eye Irritation:

A study was conducted to assess the eye irritancy potential of the test substance in one New Zealand albino female rabbit according to OECD Guideline 405 and EU Method B.5, in compliance with GLP. A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of the animal. The treated eye was washed out 24 h after administration. The untreated eye served as a control. The conjunctiva, nictitating membrane and sclera were discolored orange by the test substance up to the end of the study. The conjunctiva showed definitely injected blood vessels up to a diffuse crimson red color and swelling with the lid being half closed accompanied by clear substance coloured discharge on the day of substance administration. Based on the individual scores after 24, 48 and 72 h, the mean score was 0 for opacity of the cornea, iris and chemosis of the conjunctiva. The score was 0.33 for redness of the conjunctiva. Under the test conditions, the substance caused orange discoloration of the conjunctiva, nictitating membrane and sclera of one rabbit eye until end of the study period. Reddening was seen in the conjunctiva but this cleared within 2 days (Seeberger, 1999c).

Justification for classification or non-classification

Skin irritation:

Based on the results of a skin irritation study, the test substance does not need to be classified for skin irritation potential according to the EU CLP criteria (EC 1272/2008) as well as EU Directive 67/548/EEC.

Eye irritation:

Based on persistent discolouration of conjunctivae, nictitating membranes and iris until the end of the observation period (one animal), the test substance was classified as Xi; R41 under EU Directive 67/548/EEC. However, as no discoloration of the cornea is observed in any of the rabbit eyes, the substance does not have to be classified as causing serious damage to eyes according to the CLP Regulation (EC 1272/2008). No classification for irritating effects to the eyes is therefore required.