Pentasodium 7-(4-(4-(5-amino-4-sulfonato-2-(4-((2-(sulfonato-ethoxy)sulfonyl)phenylazo)phenylamino)-6-chloro-1,3,5-triazin-2-yl)amino-2-ureidophenylazo)naphtalene-1,3,6-trisulfonate

Endpoint summary

Description of key information

FAT 40549 was found to be non-irritant to skin and eye.

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:

22 November 1995 to 30 November 1996

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:

other: U.S. Environmental Protection Agency, Code of Federal Regulations 40, Part 798, Health Effects Testing Guidelines, Subpart E, Specific organ/Tissue toxicity, Paragraph 798.4470 "Primary dermal irritation", revised as of July 1, 1993

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: CIBACRON GELB TZ 4210 ROH LÖSUNG (LABORGETROCKNET) (FAT 40'544/A)
Description: Red powder
Batch number: TV.1
Purity/formulation: ca 90%
pH (1 g/L): 5-6
Stability of test article: stable at storage conditions: expiration date: Nov 2000
Storage conditions: At room temperature (ca 20 °C) away from sunlight.
Safety precautions: Gloves, goggles and face mask were sufficient to ensure personnel health and safety.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST SYSTEM
Test System: New Zealand White Rabbit - CRL:KBL(NZW)BR
Source: Charles River Stolzenseeweg 32-36 D-88353 Kisslegg
Age at treatment: 13 weeks
Body weight at start of acclimatization: male : 2.2 kg, females: 2.2 - 2.4 kg
Body weight at start of treatment: 2.3 kg, females: 2.2 - 2.4 kg
Acclimatization: Five days under test conditions after health examination. Only healthy animals were used for the study.
Allocation: Male no. 28, Female nos. 29 - 30
Accommodation: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
Diet: Pelleted standard Kliba 341, Batchs 91/95 and 92/95 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum.
Water: Community tap water from Itingen, ad libitum.

ENVIRONMENTAL CONDITIONS
Air-conditioned with 10 - 15 air changes per hour and continuously monitored environment with target ranges for temperature of 20 ±3 °C and for relative humidity between 40 - 70 % (values above 70 % during cleaning process possible), 12 hours artificial fluorescent light (approx. 100 Lux) /12 hours dark, music during the light period

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

bi-distilled water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 g/animal (left side only)

Duration of treatment / exposure:

4 hours

Observation period:

Viability/Mortality and Clinical Signs: Daily during the observation period.
Body Weights: At the start of acclimatization, on day 1 of test (application day) and at termination of observation.
The skin reaction was assessed at approximately 1, 24, 48 and 72 hours after the removal of the dressing, gauze patch and test article.

Number of animals:

1 male, 2 females

Details on study design:

Approximately 24 hours before treatment, the dorsal fur was clipped with an electric clipper, exposing an area of approximately 100 square centimeters (10 cm x 10 cm). The skin of the animals was examined once, 1 day prior to test article administration. Only those animals with no signs of skin injury or irritation were used in the test.

On test day 1, 0.5 g of the test article was applied to approx. 6 sq.cm of the intact skin of the clipped area. It was covered with a 3.0 cm x 3.0 cm patch of surgical gauze. The gauze was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with an elastic bandage. The test article was applied in such a position as to preclude oral ingestion after removal of the dressing.
The duration of treatment was 4 hours. Then 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.

No necropsy was performed in the animals euthanized at termination of observation.
All rabbits were euthanized by an intravenous injection of NARCOREN (Rhone Merieux GmbH, D-88471 Laupheim) into the ear vein at a dose of at least 1.0 ml/kg body weight (equivalent to 160 mg sodium pentobarbitone/kg body weight) and discarded.


EVALUATION OF SKIN REACTION
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 slight eschar formation (injuries in depth) - 4

EDEMA FORMATION
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:

No corrosive effects were noted on the treated skin of any animal at any measuring interval.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.
Orange staining by the test article of the treated skin was observed.
The body weight gain of all rabbits was within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item did not induce significant or irreversible damage to the rabbit skin.

Executive summary:

The primary skin irritation potential of the test article was investigated by topical application of 0.5 g onto 6 cm² intact dorsal skin of each of three young adult rabbits. This test was performed in accordance with OECD test guidline 404 in a GLP certified laboratory.

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 erythema and edema scores at any point were recorded to be Zero. The test article caused orange staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

In conclusion, the test substance is not be classified according to the CLP regulation (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:

experimental study

Adequacy of study:

key study

Study period:

29 November 1995 to 25 December 1995

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

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4500 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: CIBACRON GELB TZ 4210 ROH LÖSUNG (LABORGETROCKNET) (FAT 40'544/A)
Description: Red powder
Batch number: TV.1
Purity/formulation: ca 90%
pH (1 g/L): 5-6
Stability of test article: stable at storage conditions: expiration date: Nov 2000
Storage conditions: At room temperature (ca 20 °C) away from sunlight.
Safety precautions: Gloves, goggles and face mask were sufficient to ensure personnel health and safety.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test animals: New Zealand White Rabbit - CRL:KBL(NZW)BR
Animal Source: Charles River, Stolzenseeweg 32-36, D-88353 Kisslegg
Age at treatment: 13 weeks
Number of animals: 3 (1 male and 2 females)
Identification: By unique cage number and corresponding ear number.
Acclimatization: Four days under laboratory conditions after health examination. Only healthy animals were used for the study.
Body weight at start of acclimatization: male : 2.2 kg / females: 2.2 - 2.3 kg
Body weight at start of treatment: male: 2.4 kg / females: 2.3 - 2.5 kg
Standard Laboratory Conditions: Air-conditioned with 10 - 15 air changes per hour and continuously monitored environment with target ranges for temperature of 20 ±3 °C and for relative humidity between 40 - 70 % (values above 70% during cleaning process possible), 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Accommodation: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
Diet: Pelleted standard Kliba 341, Batch no. 92/95 rabbit maintenance diet (Kliba Mühlen AG, CH-4303 Kaiseraugst), ad libitum. Results of analysis for contaminants are included in the report.
Water: Community tap water from Itingen, ad libitum.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g of the test article (undiluted) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball.

Duration of treatment / exposure:

On test day 1, 0.1 g of the test article (undiluted) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball (eyes remained unrinsed). The lids were then gently held together for about one second to prevent loss of the test article. The right eye remained untreated and served as the reference control.

Observation period (in vivo):

Viability/Mortality and Clinical Signs were checked daily during the observation period. Body weights were recorded at start of acclimatisation, at start of treatment and following treatment. The eyes of each animal were examined for ocular irritation approximately 1, 24, 48 and 72 hours, as well as 7, 14 and 21 days after administration.

Number of animals or in vitro replicates:

3 (1 male and 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: No

EVALUATION OF EYE REACTIONS
CORNEAL IRRITATION
Opacity: degree of density (densest area used for assessment)
No ulceration or opacity - 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible - 1
Easily discernible translucent area, details of iris slightly obscured - 2
Nacrous area, no details of iris visible, size of pupil barely discernible - 3
Opaque cornea, iris not discernible through the opacity - 4

IRIDIC IRRITATION
Normal - 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish re action is positive) - 1
No reaction to light, hemorrhage, gross destruction (any or all of these) - 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris).
Blood vessels normal - 0
Some blood vessels definitely hyperemic (injected) - 1
Diffuse, crimson color, individual vessels not easily discernible - 2
Diffuse beefy red - 3

Chemosis: lids and/or nictitating membranes
No swelling - 0
Any swelling above normal (includes nictitating membranes) - 1
Obvious swelling with partial eversion of lids - 2
Swelling with lids about half closed - 3
Swelling with lids more than half closed - 4

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

Score:

0.78

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis 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:

CIBACRON GELB TZ 4210 ROH LÖSUNG (LABORGETROCKNET) (FAT 40'544/A) showed a primary irritation score of: 0.78, when applied to the conjunctival sac of the rabbit eye.
One (1) hour after instillation of the test article, moderate reddening and swelling of the conjunctivae were noted in all animals, as was moderate to marked watery discharge. Hyperemia of the scleral blood vessels was also evident in all animals.
Reddening and swelling of the conjunctivae tissues were reversible after 72 hours in two animals and after seven days in one animal.

Other effects:

Light orange to orange-brown staining of the conjunctivae was noted in all animals until observation on day 14. From day 14 onwards, staining of the conjunctivae was yellow. Staining of the sclera was evident in all animals on day 21 after application. The corneas of all treated animals were unaffected.

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred. No corrosion of the cornea was observed at any of the reading times. The body weight of the animals was within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance is considered non-irritant to rabbit eyes.

Executive summary:

The primary irritation potential of the test article was investigated according to method specified as per OECD test guideline 405 in a GLP certified laboratory.

0.1 g of test substance was instilled into conjuctival sac of each of three adult New Zealand 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, 14 and 21 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 averaging the individual cumulative scores at 24, 48 and 72 hours. Corneal opacity, iris and chemosis scores were zero at 24, 48 and 72h. Conjuctivae redness score was 0.78 which was reversible in 7 days.

Hyperemia of the scleral blood vessels was evident in all animals. No finding persisted longer than seven days. Light orange to orange-brown staining of the conjunctivae was noted in all animals until observation on day 14. From day 14 onwards, staining of the conjunctivae was yellow. Staining of the sclera was evident in all animals on day 21 after application. The corneas of all treated animals were unaffected.

No corrosion was observed at any of the measuring intervals. Thus, the substance is considered non-irritant to rabbit eyes.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

The primary skin irritation potential of the test article was investigated by topical application of 0.5 g onto 6 cm² intact dorsal skin of each of three young adult rabbits. This test was performed in accordance with OECD test guidline 404 in a GLP certified laboratory.

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 erythema and edema scores at any point were recorded to be Zero.The test article caused orange staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

In conclusion, the test substance is not be classified according to the CLP regulation (Regulation EC No. 1272/2008).

Eye irritation:

The primary irritation potential of the test article was investigated according to method specified as per OECD test guideline 405 in a GLP certified laboratory.

0.1 g of test substance was instilled into conjuctival sac of each of three young adult New Zealand 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, 14 and 21 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 averaging the individual cumulative scores at 24, 48 and 72 hours. Corneal opacity, iris and chemosis scores were zero at 24, 48 and 72h. Conjuctivae redness score was 0.78 which was reversible in 7 days.

Hyperemia of the scleral blood vessels was evident in all animals. No finding persisted longer than seven days. Light orange to orange-brown staining of the conjunctivae was noted in all animals until observation on day 14. From day 14 onwards, staining of the conjunctivae was yellow. Staining of the sclera was evident in all animals on day 21 after application. The corneas of all treated animals were unaffected.

No corrosion was observed at any of the measuring intervals. Thus, the substance is considered non-irritant to rabbit eyes.

Justification for selection of skin irritation / corrosion endpoint:
guidance test with GLP compliance

Justification for classification or non-classification

Based on the skin irritation / corrosion test result, the test substance shall not be classified according to the CLP regulation (Regulation EC No. 1272/2008) or DSD (Directive 67/548/EEC) for skin corrosion/irritation.

Based on the eye damage/irritation test result,the test substance shall not be classified according to the CLP regulation (Regulation EC No. 1272/2008) or DSD (Directive 67/548/EEC) for eye damage/irritation.

Data on respiratory irritation are not available.