tetrasodium 3-amino-4-{[4-({4-fluoro-6-[phenyl(2-{[2-(sulfonatooxy)ethyl]sulfonyl}ethyl)amino]-1,3,5-triazin-2-yl}amino)-2-sulfonatophenyl]diazenyl}-5-hydroxynaphthalene-2,7-disulfonate

Endpoint summary

Description of key information

No skin or eye-irritating effects have been observed in respective studies in rabbits.

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 September 28, 2004 to October 19, 2004

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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: 2.4 – 2.9 kg
- Housing: In fully air-conditioned rooms in separate cages arranged in a battery
- Diet: Ssnif K-H (V2333), ad libitum and hay (i.e., approximately 15 g daily)
- Water: Tap water in plastic bottles, ad libitum
- Acclimation period: 1 week under study conditions
-Animal identification: Ear tags

ENVIRONMENTAL CONDITIONS
- Temperature: 18±3°C (except short lasting deviations due to disturbances of air condition)
- Humidity: 50±20% (except short lasting deviations due to disturbances of air condition)
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: September 28, 2004 to October 19, 2004

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

deionized

Controls:

yes, concurrent no treatment

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 h

Observation period:

30-60 minutes, 24, 48, 72 h and 7 and 14 d after removal of the patches

Number of animals:

3 females

Details on study design:

ROUTE OF ADMINISTRATION: Dermal

FREQUENCY OF ADMINISTRATIONS: Single dose

PREPARATION OF THE TEST SUBSTANCE
Each animal was treated with 0.5 g (i.e., 500 mg) of test substance pasted with 0.35 mL of deionized water.

TEST PROCEDURE
An initial test was performed using one animal.

As no corrosive effects were observed in the initial test a confirmatory test with two further animals was conducted.

About 24 h before the start of the study the hair in the dorsal region of the body of the 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 mg of test substance pasted with 0.35 mL deionized water. The test substance was evenly distributed 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. Because of persistent discolorations by the test substance 72 h after removal of the patches, additional readings were performed after 7 and 14 d.

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.....................................................................................................................9
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

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

Irritant / corrosive response data:

30–60 minutes up to 7 or 14 d after removal of the patches the treated skin of the animals showed small or large light pink substance discolored areas. No signs of irritations were noted.

Other effects:

No clinical signs of systemic toxicity were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test substance was considered to be non-irritating to skin in an in vivo study with rabbits.

Executive summary:

A study was performed to assess the skin irritation potential of the test substance in female Crl:KBL(NZW)BR rabbits according to OECD Guideline 404, EPA OPPTS 870.2500 and EU Method B.4, in compliance with GLP.

500 mg of test substance pasted with 0.35 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 from the skin with warm tap water. Animals were scored for irritation reactions after 30 - 60 minutes, and at 24, 48, 72 h and 7 and 14 d.

No severe signs of irritation were observed in treated animals during the whole observation period. Further, overall mean scores for dermal irritation were calculated to be 0.00 for erythema and eschar formation and 0.00 for edema formation. In addition, no clinical signs of systemic toxicity were observed.

 

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 October 12, 2004 to November 02, 2004

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:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, 88353 Kißlegg
- Body weight at treatment: 2.8 kg
- Housing: In fully air-conditioned rooms in separate cages arranged in a battery
- Diet: Ssnif K-H (V2333), ad libitum and hay (i.e., approximately 15 g daily)
- Water: Tap water in plastic bottles, ad libitum
- Acclimation period: 1 week under study conditions
- Animal identification: Ear tags

ENVIRONMENTAL CONDITIONS
- Temperature: 18±3°C (short lasting deviations are permissible, e.g., during cleaning processes)
- Humidity: 50±20% (short lasting deviations are permissible, e.g., during cleaning processes)
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: October 12, 2004 to November 02, 2004

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 and 21 d following administration of the test substance

Number of animals or in vitro replicates:

1 female

Details on study design:

ROUTE OF ADMINISTRATION: Conjunctival

FREQUENCY OF ADMINISTRATIONS: Single dose

PREPARATION OF THE TEST SUBSTANCE:
The animal was treated with 0.1 g (i.e., 100 mg) of test substance

TEST PROCEDURE
An initial test was performed using one animal.

As the test substance showed potential risk of serious damage to the eye only one animal was used in the study.

About 24 h before the start of the study both eyes of the animal was 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.

0.1 g of 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 37°C. The eye was also washed out at designated examinationtimes 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 were 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:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Max. score:

3

Remarks on result:

not determinable

Remarks:

Due to the violet discoloration of the eye the animal could not be assessed for redness of conjunctiva

Irritant / corrosive response data:

From 1 h up to 1 d after application the animal showed slight swellings of lids. The irritations were attended by eye discharge discolored by the test substance (i.e., violet) 1 h after administration. Additionally, conjunctiva, nictitating membrane and iris were discolored by the test substance (i.e., violet) up to the end of the study (i.e., Day 22).

Other effects:

No clinical signs of systemic toxicity were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the substance caused violet discoloration of the conjunctiva, nictating membrane and iris of one rabbit eye until end of the study period. However, no signs of irritation was found and the substance is therefore not considered irritating to the eye according to CLP criteria.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in female NZW White (Crl:KBL(NZW) rabbits 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 one animal. The treated eye was washed out 24 h after administration. The untreated eye served as a control.

The conjunctiva, nictitating membrane and iris were discolored (violet) by the test substance up to the end of the study (Day 22). Overall mean scores for eye irritation were calculated to be 0.0 for iris and opacity of cornea and 0.33 for chemosis of conjunctiva. However, due to the violet discoloration of the eye, the animal could not be assessed for redness of conjunctiva. No clinical signs of systemic toxicity were seen.

Under the test conditions, the substance caused violet discoloration of the conjunctiva, nictating membrane and iris of one rabbit eye until end of the study period.

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 Crl:KBL(NZW)BR rabbits according to OECD Guideline 404, EPA OPPTS 870.2500 and EU Method B.4, in compliance with GLP.500 mg of test substance pasted with 0.35 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 from the skin with warm tap water. Animals were scored for irritation reactions after 30 - 60 minutes, and at 24, 48, 72 h and 7 and 14 d.No severe signs of irritation were observed in treated animals during the whole observation period. Further, overall mean scores for dermal irritation were calculated to be 0.00 for erythema and eschar formation and 0.00 for edema formation. In addition, no clinical signs of systemic toxicity were observed.Under the test conditions, the test substance was considered to be non-irritating to skin.

Eye irritation:

A study was conducted to assess the eye irritancy potential of the test substance in female NZW White (Crl:KBL(NZW) rabbits 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 one animal. The treated eye was washed out 24 h after administration. The untreated eye served as a control. The conjunctiva, nictitating membrane and iris were discoloured (violet) by the test substance up to the end of the study (Day 22). Overall mean scores for eye irritation were calculated to be 0.0 for iris and opacity of cornea and 0.33 for chemosis of conjunctiva. However, due to the violet discoloration of the eye, the animal could not be assessed for redness of conjunctiva. No clinical signs of systemic toxicity were seen. Under the test conditions, the substance caused violet discoloration of the conjunctiva, nictitating membrane and iris of one rabbit eye until end of the study period.

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).

Eye irritation:

Based on the results of a eye irritation study, the test substance does not need to be classified for eye irritation potential according to the EU CLP criteria (EC 1272/2008).