Tetrasodium 2-[[8-[[3-[(5-chloro-2,6-difluoro-4-pyrimidinyl)amino]benzoyl]amino]-1-hydroxy-3,6-disulphonato-2-naphthyl]azo]naphthalene-1,5-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:

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

Adequacy of study:

key study

Study period:

September from 14 to 23, 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

The read across approach is detailed into the document attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted JuIy 17,1992

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes F-01400 Chatillon sur Chalaronne / France.
- Age at study initiation: 14 weeks old male and 15 weeks old females.
- Weight at study initiation: male ca 2.6 kg; females ca 2.7 kg.
- Housing: individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no.65/99).
- Water: community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 5 days, under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
- Health check: approximately four days before treatment, a flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm x 10 cm). The skin of the animals was examined approximately 24 hours before treatment and regrown fur was again clipped. The fur was again slightly clipped in two animals (Nos. 80 - 81) just prior to the application. Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Relative humidity: 40 - 70 %
- Air changes: 10-15 air changes per hour.
- Photoperiod: 12 hour light/dark cycle.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 g/animal (left side only).
The test article was moistened with bi-distilled water before application.

Duration of treatment / exposure:

4 hours.

Observation period:

72 hours

Number of animals:

1 male and 2 females

Details on study design:

TEST SITE
- Area of exposure: on the day of treatment test item was setted on a surgical garuze patch (ca.2.5 cm x 2.5 cm). The gauze patch was applied to approximately 6 cm2 of the intact skin of the clipped area.
- Type of wrap if used: the patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing: the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4 hours.

OBSERVATIONS
- Viability/Ivlortality: daily from delivery of the animals to the termination of test.
- Clinical signs: daily from delivery of the animals to the termination of test.
- Body weights: at start of acclimatization, on the day of application and at termination of observation.

OBSERVATION TIME POINTS
The skin reaction was assessed at approximately 1, 24, 48 and 72 hours after the removal of the dressing, gauze patch and test article.

SCORING SYSTEM:
The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992.
If evident, corrosive or staining properties of the test article were described and recorded.

ERYTHEMA AND ESCHAR FORMATION
No erythema 0
Very slight erythema 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) or eschar formation (injuries in depth preventing erythema reading 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 (edges raised approximately 1 mm) 3
Severe oedema (raised more than I mm and extending beyond the 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:

Application of the test article to healthy intact rabbit skin resulted in a primary irritation score of 0.00.
Local signs (mean values from 24 to 72 hours) consisted of grade 0.00 erythema and grade 0.00 oedema. No signs of irritation were observed.
Light red staining by the test article of the treated skin was observed.
No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

VIABILITY/MORTALITY AND CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.

BODY WEIGHTS
The body weight of all rabbits were considered to be within the normal range of variability.

Individual reactions

Animal	Reaction	1 hr	24 hrs	48 hrs	72 hrs
79 male	Erythema	0	0	0	0
	Oedema	0	0	0	0
80 female	Erythema	0	0	0	0
	Oedema	0	0	0	0
81 female	Erythema	0	0	0	0
	Oedema	0	0	0	0

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Not skin irritating

Executive summary:

The primary skin irritation potential of the test article was investigated by topical application of 0.5 g to 6 cm2 intact dorsal skin of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours after removal of the dressing. The scores of each animal at the following reading times (24, 48, 72 hours) were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by totaling the individual cumulative scores at 24, 48 and 72 hours and then dividing by the number of data points. The primary irritation score was 0.00 (max. 8.0).

Local signs (mean values from 24 to 72 hours) consisted of grade 0.00 erythema and grade 0.00 oedema. The test article caused light red staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

Conclusion

None of the animals showed any observable response to treatment throughout the 72 hours observation period, thus it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.

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:

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

Adequacy of study:

key study

Study period:

From September 21 to October 04, 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

The read across approach is detailed into the document attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted February 24, 1987

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes F-01400 Chatillon sur Chalaronne / France.
- Age at study initiation: 15 weeks old.
- Weight at study initiation: male ca 2.7 kg; females ca 2.7-2.8 kg.
- Housing: individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no.65/99).
- Water: community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 5 days, under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
- Health check: the eyes of the animals were examined one day prior to test article administration. Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of the results were not used in the test.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Relative humidity: 40 - 70 %
- Air changes: 10-15 air changes per hour.
- Photoperiod: 12 hour light/dark cycle.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g/animal (left eye only).

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

OBSERVATIONS
- Viability/Mortality: daily from delivery of the animals to the termination of test.
- Clinical signs: daily from delivery of the animals to the termination of test.
- Body weights: at start of acclimatization, on the day of application and at termination of observation.

TOOL USED TO ASSESS SCORE: eye examinations were made with a Varta Cliptrix diagnostic-lamp (A. Riegger, Basel / Switzerland).

SCORING SYSTEM
The ocular reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC. July 31,1992 at approximately 1, 24, 48 and72 hours, as well as 7 days after administration.
When present, corrosion of and/or staining of sclera and cornea by the test article were recorded and reported.

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: 7 days

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 24 hours

Irritant / corrosive response data:

Application of the test article to healthy rabbit conjunctivae resulted in a primary irritation score of 0.56.
Hyperemic conjunctival blood vessels were noted in all animals at the 24-hour examination and persisted through 72 hours in one animal only. This latter was observed with a slight swelling of the conjunctivae at the 1-hour examination and with a slightly reddened sclera from the 24- to the 72-hour examination. Slight to moderate watery discharge was also observed in all animals at the 1-hour examination.
All eye reactions were reversible within 7 days after treatment.
Reddish-brown remnants in eye or conjunctival sac and around eye and/or on lids were observed until the 72-hour reading.
No corrosion of the cornea was observed at any of the reading times.

Other effects:

VIABILTY/MORTALITY AND CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.

BODY WEIGHTS
The body weight of all rabbits were considered to be within the normal range of variability.

Individual reactions

Animal N.	Reaction	After					Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs	7 days
76 male	Corneal opacity	0	0	0	0	0	0.00
77 female	Corneal opacity	0	0	0	0	0	0.00
78 female	Corneal opacity	0	0	0	0	0	0.00
76 male	Iris	0	0	0	0	0	0.00
77 female	Iris	0	0	0	0	0	0.00
78 female	Iris	0	0	0	0	0	0.00
76 male	Coniunctivae redness	0	1	0	0	0	0.33
77 female	Coniunctivae redness	0	1	0	0	0	0.33
78 female	Coniunctivae redness	0	1	1	1	0	1.00
76 male	Coniunctivae chemosis	0	0	0	0	0	0.00
77 female	Coniunctivae chemosis	1	0	0	0	0	0.00
78 female	Coniunctivae chemosis	1	0	0	0	0	0.00

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Non irritating

Executive summary:

The primary irritation potential of test item was investigated by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7 days after test article application. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by totaling the individual cumulative scores at 24, 48 and 72 hours and then dividing the resulting total by the number of data points. The primary irritation score was 0.56 (max. 13). Hyperemic conjunctival blood vessels were noted in all animals at the 24-hour examination and persisted through 72 hours in one animal only. This latter was observed with a slight swelling of the conjunctivae at the 1-hour examination.

Reddish-brown rernnants in eye or conjunctival sac and around eye and/or on lids were observed until the 72-hour reading. All eye reactions were reversible within 7 days after treatment. No corrosion was observed at any of the measuring intervals.

Conclusion

The mean values from gradings 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 chemosis, in all the tested animals. All the reactions recorded were reversible within 7 days. Therefore, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC) No 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

SKIN IRRITATION

There are no information about the skin irritation potential of Reactive Red 147, thus the available information on the structural analogous Similar Substance 01 have been taken into consideration; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID section 13).

The primary skin irritation potential of the Similar Substance 01 was investigated by topical application of 0.5 g to 6 cm² intact dorsal skin of each of three young adult rabbits. The duration of treatment was four hours. The primary irritation score was calculated by totaling the individual cumulative scores at 24, 48 and 72 hours and then dividing by the number of data points. The primary irritation score was 0.00 (max. 8.0). Local signs (mean values from 24 to 72 hours) consisted of grade 0.00 erythema and grade 0.00 oedema. The test article caused light red staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

EYE IRRITATION

There are no information about the eye irritation potential of Reactive Red 147, thus the available information on the structural analogous Similar Substance 01 have been taken into consideration; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID section 13).

The primary irritation potential of Similar Substance 01 was investigated by instillation of 0.1 g into one eye of each of three young adult rabbits. The treated eyes were not rinsed after application. The primary irritation score was calculated by totaling the individual cumulative scores at 24, 48 and 72 hours and then dividing the resulting total by the number of data points. The primary irritation score was 0.56 (max. 13). Hyperemic conjunctival blood vessels were noted in all animals at the 24-hour examination and persisted through 72 hours in one animal only. This latter was observed with a slight swelling of the conjunctivae at the 1-hour examination. Reddish-brown rernnants in eye or conjunctival sac and around eye and/or on lids were observed until the 72-hour reading. All eye reactions were reversible within 7 days after treatment. No corrosion was observed at any of the measuring intervals.

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 gradings 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 gradings 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 chemosis, in all the tested animals. All the reactions recorded were reversible within 7 days.

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