2-Naphthalenesulfonic acid, 4-hydroxy-3-[2-[2-methoxy-5-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-7-[(sulfomethyl)amino]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:5)

Endpoint summary

Description of key information

The test substance is not considered to be irritating to skin and eyes.

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 Mar. 22, 2005 to Apr. 29, 2005

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
- Weight at study initiation: 2.2–2.9 kg
- Housing: in fully air-conditioned rooms in separate cages arranged in a battery
- Diet: ssniff® K-H (V2333), ad libitum and hay (approximately 15 g daily)
- Water: tap water in plastic bottles, ad libitum
- Acclimation period: 1 week under study conditions
- Temperature: 18±3°C
- Humidity: 50±20%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From Mar. 22, 2005 to Apr. 29, 2005

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 500 mg

VEHICLE
- Amount applied: 0.3 mL

Duration of treatment / exposure:

4 h

Observation period:

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 irritations 72 h after removal of the patches, additional readings were performed after 7 d with one animal. First animal exposed which had servere test item related skin discoloration was observed until the discoloration had disappeared (Day 15) to assess reversibility of discoloration.

Number of animals:

4

Details on study design:

TEST SITE
- Preparaton of test site: About 24 h before the start of the study the hair in the dorsal region of the body of the 4 rabbits was removed with an electric clipper over an area of about 25 cm². Only animals with intact skin were used.
- Type of wrap if used: Semi-occlusive

REMOVAL OF TEST SUBSTANCE
- Washing: Warm tap water.
- Time after start of exposure: 4 h

SCORING SYSTEM: Draize

Overall study design:
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 at first. A 4th animal had to be used for valid assessment because one animal was not assessable for erythema in the first study week due to test substance related discolorations. About 24 h before the start of the study the hair in the dorsal region of the body of the 4 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 of test substance formed into a paste-like consistency with 0.3 mL of deionized water.The test substance was evenly distributed over the whole surface of a 2.5x2.5 cm cellulose patch on a piece of surgical plaster (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 irritations 72 h after removal of the patches, additional readings were performed after 7 d with one animal. First animal which showed severe test substance related skin discoloration was observed until the discoloration had disappeared (Day 15) to assess reversibility of discoloration. Erythema, eschar formation and edema were evaluated numerically according to the score of Draize. All other changes of the skin were recorded.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0.78

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0.56

Max. score:

4

One (1^st) animal was not assessable for erythema in the first study week because of test substance related skin discoloration. Therefore, a 4^thanimal was treated in addition to the remaining animals. 1^st animal was not included in overall assessment.

30-60 minutes up to 24 or 72 h after decontamination the animals 2^ndand 4^thshowed sporadically very slight up to well-defined erythema. In case of 2^ndanimal, erythema was not assessable due to the discoloration caused by the test item 30-60 minutes after decontamination.

Very slight up to slight edema were observed 24 h up to 72 h after decontamination in second animal. Additionally, the skin surface of the 2^ndanimal was dry, rough and scaled. 7 d after decontamination the irritations were reversed. 3^rd animal showed no irritations.

Interpretation of results:

GHS criteria not met

Conclusions:

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

Executive summary:

A study was conducted to assess the skin irritation potential of the test substance in New Zealand White rabbits according to EU Method B.4 and OECD Guideline 404, in compliance with GLP.

0.5 g of test substance was mixed with 0.3 mL of deionized water to form a paste-like consistency and applied as a single dose on the clipped dorsal surface of the test animals. As no corrosive effects were observed in the initial test, a confirmatory study with two further animals was conducted. A 4^thanimal had to be added because one animal was not assessable for erythema in the first study week due test substance-related discoloration. The substance was evenly distributed over a surface of 2.5 x 2.5 cm using a semi-occlusive patch. The substance was washed with warm water after a 4 h exposure period. Examinations of the skin took place 30-60 minutes as well as 24, 48 and 72 h after removal of the patches. Because of persistent irritation 72 h after patch removal, additional readings were performed after 7 d with one animal. The animal which showed severe skin discoloration was observed until this had disappeared (Day 15). Erythema, eschar formation and edema were evaluated according to the score of Draize.

No clinical signs of systemic toxicity were observed. From 30-60 minutes up to 24 or 72 h after washing, two animals showed sporadically very slight up to well-defined erythema. Additionally, the skin surface was dry, rough and scaled. 7 d after washing, the irritation was reversed. A third animal showed no irritation.

Under the conditions of the study, the test substance was found 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 Apr. 19, 2005 to May 10, 2005

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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, 88353 Kißlegg
- Weight at study initiation: 3.63 kg
- Housing: in fully air-conditioned rooms in separate cages arranged in a battery
- Diet: ssniff K-H (V2333), ad libitum and hay (approx. 15 g daily)
- Water: tap water in plastic bottles, ad libitum
- Acclimation period: 1 week under study conditions

ENVIRONMENTAL CONDITIONS
- Temperature: 18 ± 3°C
- Humidity: 50 ± 20 %
- Air changes:

IN-LIFE DATES: From: Apr. 19, 2005 To: May 10, 2005

Vehicle:

not specified

Remarks:

No vehicle used

Controls:

other: the untreated right eye served as a control

Amount / concentration applied:

TEST MATERIAL
- Amount applied : 0.1 g

Duration of treatment / exposure:

24 h

Observation period (in vivo):

21 d

Number of animals or in vitro replicates:

1

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: 24 h after start of exposure with isotonic saline at approx. 37 °C

SCORING SYSTEM: as per OECD Guideline

TOOL USED TO ASSESS SCORE: fluorescein and UV light

Irritation parameter:

cornea opacity score

Basis:

animal #1

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

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Irritant / corrosive response data:

From 1 h up to 72 h after administration the conjunctiva showed definitely injected blood vessels up to a crimson red color and up to 24 h after treatment some swelling above normal up to obvious swelling with partial eversion of lids. 24 h after application the nictitating membrane of the animal was discolored by the test item up to the end of the observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the substance caused discoloration of the nictitating membrane of rabbit eye until end of the study period. No other signs of irritation were observed.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in one NZW White 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.

Overall mean scores for eye irritation were calculated to be 0.0 for iris and opacity of cornea, 1.33 for redness of conjunctiva and 0.33 for chemosis of conjunctiva. 24 h after the application, the nictitating membrane was discoloured by the test substance up to the end of the study. No clinical signs of systemic toxicity were seen. Body weight gain of the treated animal was not affected.

Due to the fact that a classification of R41 under DSD/DPD was warranted due to the non-reversible discolouration of the nictitating membrane, no further animals were tested due to animal welfare reasons. However, as only the conjunctivae of the rabbit eye were discoloured. And no other signs of eye irritation were found, following the criteria of Regulation (EU) 1272/2008, no classification applies in terms of serious eye irritation/eye damage.

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 conducted to assess the skin irritation potential of the test substance in New Zealand White rabbits according to EU Method B.4 and OECD Guideline 404, in compliance with GLP. 0.5 g of test substance was mixed with 0.3 mL of deionized water to form a paste-like consistency and applied as a single dose on the clipped dorsal surface of the test animals. As no corrosive effects were observed in the initial test, a confirmatory study with two further animals was conducted. A 4^thanimal had to be added because one animal was not assessable for erythema in the first study week due test substance-related discoloration. The substance was evenly distributed over a surface of 2.5 x 2.5 cm using a semi-occlusive patch. The substance was washed with warm water after a 4 h exposure period. Examinations of the skin took place 30-60 minutes as well as 24, 48 and 72 h after removal of the patches. Because of persistent irritation 72 h after patch removal, additional readings were performed after 7 d with one animal. The animal which showed severe skin discoloration was observed until this had disappeared (Day 15). Erythema, eschar formation and edema were evaluated according to the score of Draize. No clinical signs of systemic toxicity were observed. From 30-60 minutes up to 24 or 72 h after washing, two animals showed sporadically very slight up to well-defined erythema. Additionally, the skin surface was dry, rough and scaled. 7 d after washing, the irritation was reversed. A third animal showed no irritation. Under the conditions of the study, the test substance was found to be non-irritating to skin (Kauffmann, 2005d).

Eye irritation:

A study was conducted to assess the eye irritancy potential of the test substance in one New Zealand White 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. Overall mean scores for eye irritation were calculated to be 0.0 for iris and opacity of cornea, 1.33 for redness of conjunctiva and 0.33 for chemosis of conjunctiva. 24 h after the application, the nictitating membrane was discolored by the test substance up to the end of the study. No clinical signs of systemic toxicity were seen. Body weight gain of the treated animal was not affected. Under the test conditions, the test substance was found to be non-irritating to the eyes (Kaufmann, 2005e).

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 this endpoint according to the EU CLP criteria (EC 1272/2008) as well as EU Directive 67/548/EEC.

Eye irritation:

Based on persistent staining of the nictitating membrane in one animal at the end of the observation period, the test substance is classified as 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 warrant classification according to EU CLP criteria (EC No 1272/2008).