Trisodium 7-[[4-chloro-6-[ethyl[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-4-hydroxy-3-[(4-methoxy-2-sulphonatophenyl)azo]naphthalene-2-sulphonate

Endpoint summary

Description of key information

The test item is not irritating to skin or 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 July 12, 1982 to July 26, 1982

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:

Abraded and intact dorsal skin (previously clipped) of six New-Zealand white rabbits was exposed to 500 mg test substance for 24 h under occlusive conditions. At the end of the exposure period, the treated skin area was cleaned and irritation/corrosion was assessed at 24, 48 and 72 h and at 1 and 2 weeks according to Draize scoring system.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Origin: Nihon Dobutsu Co., Osaka
Body weight: 2.28 - 3.17 kg
Temperature and relative humidity: 24+/-2°C and 60+/-10%, respectively
Lighting time: 12 h daily
Food: CG-3 type (100 g/day)
Water: tap water, ad libitum

Type of coverage:

occlusive

Preparation of test site:

other: abraded or intact

Remarks:

previously clipped

Vehicle:

physiological saline

Amount / concentration applied:

500 mg of the test substance (slightly moistened with physiological saline) on 1 x 1 inch lint patch

Duration of treatment / exposure:

24 h

Observation period:

14 d

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: 4x (2.5 x 2.5 cm²) 2 shaved and 2 abraded sites per anmal
- % coverage:
- Type of wrap if used: Blenderm® surgical tape, 3M Co., Saint Paul, U.S.A.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): at the end of exposure period, the patches were removed from the skin and the treated area was wiped to remove the remaining test material.
- Time after start of exposure:24 h

OBSERVATION TIME POINTS
24, 48 and 72 hrs and 1 and 2 weeks after application

SCORING SYSTEM: Skin reactions of erythema and edema were scored 24, 48 and 72 h and 1 and 2 weeks after application according to the Draize scoring system.
- Method of calculation: Primary irritation score= Sum of (Total scores of erythema and edema at 24 and 72 hrs in 2 intact and 2 abraded skins / 8) for all 6 rabbits / 6

Irritation parameter:

erythema score

Remarks:

shaved skin

Basis:

mean

Remarks:

of 6 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

shaved skin

Basis:

mean

Remarks:

of 6 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

primary dermal irritation index (PDII)

Remarks:

shaved and abraded skin

Basis:

mean

Remarks:

of 6 rabbits

Time point:

other: 24 and 72 h

Score:

0.17

Max. score:

8

Reversibility:

fully reversible within: 14 d

Irritant / corrosive response data:

The the test material produced very slight erythema and slight edema at 24 hrs after application in one abraded skin patch, any other skin irritant reactions were not observed. The erythema gradually developed into eschar at 3 days, and disappeared within 10 days. The edema have gradually disappeared within 14 days. The primary irritation score was 0.17.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the substance was not considered to be irritating to skin.

Executive summary:

A study was conducted to determine the in vivo skin irritation potential of the substance according to an internal method of the laboratory. Clipped abraded and intact dorsal skin (ca. 1 x 1 inch) of six New-Zealand white rabbits was exposed to 500 mg of the test substance (slightly moistened with physiological saline) for 24 h under occlusive conditions. At the end of the exposure period, the treated skin area was carefully cleaned. Skin irritation/corrosion was assessed at 24, 48 and 72 h as well as at 1 and 2 weeks after bandage removal according to the Draize scoring system. The test substance produced some grade of erythema and eschar and very slight edema at 24 h after application only in one abraded skin. These changes disappeared within 14 d after application. Under the study conditions, the substance was not considered to be irritating to skin (Hosokawa, 1982).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

From July 12, 1982 to July 26, 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Remarks:

in unwashed eyes the duration of te observation period is insufficient; in washed eyes, the study is acceptable

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

Nine New-Zealand white rabbits received 100 mg of the test substance in the conjunctival sac of one eye. The other eye served as negative control. The treated eyes of 6 animals remained unwashed and eyes of the other 3 animals were washed after application. Signs of ocular irritation were examined at 1, 24, 48, 72 and 96 h and at 1 and 2 weeks after treatment. The grading and scoring of irritating reactions was performed in accordance with the Draize scale.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Origin: Nihon Dobutsu Co., Osaka
Body weight: 2.28 - 3.17 kg
Temperature and relative humidity: 24+/-2°C and 60+/-10%, respectively
Lighting time: 12 h daily
Food: CG-3 type (100 g/day)
Water: tap water, ad libitum

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg on the everted lower lid

Duration of treatment / exposure:

Unwashed group: NA
Washed group: 30 seconds

Observation period (in vivo):

14 d

Number of animals or in vitro replicates:

Unwashed group: 6
Washed group: 3

Details on study design:

Nine New-Zealand white rabbits received 100 mg of the test substance in the conjunctival sac of one eye. The other eye remained untreated and served as negative control. The treated eyes of 6 animals remained unwashed and eyes of the other 3 animals were washed after application. Reading of ocular lesions was conducted at 1, 24, 48, 72 and 96 h and at 1 and 2 weeks after treatment. The grading and scoring of irritating reactions was performed in accordance with the Draize scale.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): in 3 animals for 1 minute with 300 mL lukewarm water
- Time after start of exposure: 30 seconds

SCORING SYSTEM: Draize

Irritation parameter:

cornea opacity score

Remarks:

washed

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Remarks:

unwashed

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

0.83

Max. score:

4

Reversibility:

not fully reversible within: 14 d

Irritation parameter:

iris score

Remarks:

washed

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Remarks:

unwashed

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

washed

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

unwashed

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Remarks:

washed

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Remarks:

unwashed

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

0.17

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test substance produced some grade of irritant reactions such as cornea opacity and hyperemia, chemosis and discharge in conjunctiva in unwashed eyes at 1 or 24 h after application. In the washed eyes, no irritant reaction was observed in the animals. The mean total score of these irritant reactions for the unwashed group was maximum at 24 h after application and the score was 11.2. Although slight cornea opacity remained in 4/6 unwashed animals after 14 d, other irritant reactions disappeared within 2 to 7 d after application in all remaining animals.

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the study conditions, the substance was considered not to be irritating to eyes if the substances is washed out of the eyes. If the substance is not washed out, slight cornea opacity remained for 14 days in 4 of 6 rabbits showing a tendency for reversibility in the areas affected.

Executive summary:

A study was conducted to determine the eye irritation potential of the substance according to an internal method of the laboratory. Nine New-Zealand white rabbits received 100 mg of the test substance in the conjunctival sac of one eye. The other eye remained untreated and served as negative control. The treated eyes of 6 animals remained unwashed and eyes of the other 3 animals were washed after application. Signs of ocular irritation were examined at 1, 24, 48, 72 and 96 h and at 1 and 2 weeks after treatment. The grading and scoring of irritating reactions was performed in accordance with the Draize scale. The test substance produced irritant reactions such as corneal opacity and hyperemia, chemosis and discharge in the conjunctiva of unwashed eyes at 1 and 24 h after application. In the washed eyes, no irritant reaction was observed in the animals. The mean total score of these irritant reactions for the unwashed group was 11.2 at 24 h. Slight corneal opacity remained in 4 out of 6 unwashed animals at 14 d, other irritant reactions disappeared within 2 to 7 d after application in all remaining animals. (Hosokawa, 1982).

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

RA study

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0.56

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0.11

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

One hour up to three days after administration the conjunctivae of the animals showed sporadically definitely injected blood vessels up to a deeper crimson red
colour. One hour, in one case up to one day after administration slight swellings up to obvious swellings were noted. Additionally, serous eye discharge discoloured by the compound and orange discoloured nictitating membranes were observed. Seven days after administation all signs of irritation were disappeared.
.

Interpretation of results:

GHS criteria not met

Conclusions:

Testing of the substance for primary eye irritation in the rabbit showed that the substance is not irritating to eye

Executive summary:

A study was conducted to assess the eye irritancy potential of the read-across substance in New Zealand albino 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 three animals. The treated eyes were washed out 24 h after administration. The untreated eye served as a control. The eyes were examined 1, 24, 48 and 72 hours after administration of the test substance. At 24 and 72 hours and after 7 days, the eyes 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 according to Draize. All other changes or toxic effects were recorded. Since effects were still present in the eyes after 72 hours, further examinations were carried out after 7 days.

One hour up to three days after administration the conjunctivae of the animals showed sporadically definitely injected blood vessels up to a deeper crimson red colour. One hour, in one case up to one day after administration slight swellings up to obvious swellings were noted. Additionally, serous eye discharge discoloured by the compound and orange discoloured nictitating membranes were observed. Seven days after administration all signs of irritation were disappeared.

In conclusion, testing of the test substance for primary eye irritation in the rabbit showed that the substance is not irritating to eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation, in vivo:

A study was conducted to determine the in vivo skin irritation potential of the substance according to an internal method of the laboratory. Clipped abraded and intact dorsal skin (ca. 1 x 1 inch) of six New-Zealand white rabbits was exposed to 500 mg of the test substance (slightly moistened with physiological saline) for 24 h under occlusive conditions. At the end of the exposure period, the treated skin area was carefully cleaned. Skin irritation/corrosion was assessed at 24, 48 and 72 h as well as at 1 and 2 weeks after bandage removal according to the Draize scoring system. The test substance produced some grade of erythema and eschar and very slight edema at 24 h after application only in one abraded skin. These changes disappeared within 14 d after application. Under the study conditions, the substance was not considered to be irritating to skin (Hosokawa, 1982).

 

Eye irritation, in vivo:

A study was conducted to determine the eye irritation potential of the substance according to an internal method of the laboratory. Nine New-Zealand white rabbits received 100 mg of the test substance in the conjunctival sac of one eye. The other eye remained untreated and served as negative control. The treated eyes of 6 animals remained unwashed and eyes of the other 3 animals were washed after application. Signs of ocular irritation were examined at 1, 24, 48, 72 and 96 h and at 1 and 2 weeks after treatment. The grading and scoring of irritating reactions was performed in accordance with the Draize scale. The test substance produced irritant reactions such as corneal opacity and hyperaemia, chemosis and discharge in the conjunctiva of unwashed eyes at 1 and 24 h after application. In the washed eyes, no irritant reaction was observed in the animals. The mean total score of these irritant reactions for the unwashed group was 11.2 at 24 h. Slight corneal opacity remained in 4 out of 6 unwashed animals at 14 d, other irritant reactions disappeared within 2 to 7 d after application in all remaining animals. Under the study conditions, the substance was considered not to be irritating to eyes if the substances is washed out of the eyes. If the substance is not washed out, slight cornea opacity remained for 14 days in 4 of 6 rabbits showing a tendency for reversibility in the areas affected (Hosokawa, 1982). As the observation period was too short for unwashed eyes to evaluate the complete reversibility of the test substance, the study cannot be used for classification of the substance.

 

A study was conducted to assess the eye irritancy potential of the structural analogue in New Zealand albino 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 three animals. The treated eyes were washed out 24 h after administration. The untreated eye served as a control. The eyes were examined 1, 24, 48 and 72 hours after administration of the test substance. At 24 and 72 hours and after 7 days, the eyes 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 according to Draize. All other changes or toxic effects were recorded. Since effects were still present in the eyes after 72 hours, further examinations were carried out after 7 days. One hour up to three days after administration the conjunctivae of the animals showed sporadically definitely injected blood vessels up to a deeper crimson red colour. One hour, in one case up to one day after administration slight swellings up to obvious swellings were noted. Additionally, serous eye discharge discoloured by the compound and orange discoloured nictitating membranes were observed. Seven days after administration all signs of irritation disappeared. In conclusion, testing of the test substance for primary eye irritation in the rabbit showed that the substance is not irritating to eye

Justification for classification or non-classification

An in vivo eye irritation study showed grades of corneal opacity in majority of the test rabbits in which the treated eyes were not washed. Although the corneal opacity scores were lower than the cut-off values mandated by the EU CLP (EC 1272/2008) criteria and showed reversibility, the effects were not completely reversed by the end of the 14 day observation period and four out of six animals presented a score of 1 at study end. Since the observation period was not prolonged to 21 days as requested by the guidelines to ensure complete reversibility of the effects, the study part with unwashed eyes cannot be used for classification of the test substance.

Thus, a study with the structural analogue was further taken into account in a weight of evidence approach with the test substance.

In this study, all irritant effects were reversed 7 days after test substance administration.

Based on the study part in washed eyes and the study with the read across substance, it is therefore concluded that the test substance is not irritating to eyes.

Therefore, based on the in vivo skin and eye irritation studies with the substance, no classification is warranted according to EU CLP (EC 1272/2008) criteria.