Reaction mass of pentasodium 2-{[4-chloro-6-(ethyl{3-[(2-sulfonatoethyl)sulfonyl]phenyl}amino)-1,3,5-triazin-2-yl]amino}-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate and tetrasodium 2-[(4-chloro-6-{ethyl[3-(vinylsulfonyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate

Endpoint summary

Description of key information

The test substance is not irriatating to rabbit's skin as well as 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:

11 January 2006 to 08 February 2006

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:

not specified

Qualifier:

according to guideline

Guideline:

other: Commission Directive 2004/73/EC, B.4. "Acute Toxicity: Dermal Irritation/Corrosion", April 29, 2004.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Storage: At room temperature at about 20°C in a desiccator (hygroscopic)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Test system: Young Adult New Zealand White Rabbit, SPF
- Source: Charles River Deutschland GmbH Stolzenseeweg 32-36 D-88353 Kisslegg / Germany
- Age at treatment: 13 weeks (male), 13-14 weeks (females)
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 57/05, Provimi Kliba AG) were provided for gnawing.
- Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 74/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water: Community tap water from Füllinsdorf, ad libitum.
- Acclimation: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
- Allocation: Male No. 30, Female Nos. 31 and 32

ENVIRONMENTAL CONDITIONS
Air-conditioned with ranges for room temperature 17-23 °C, relative humidity 30-70 % and approximately 10-15 air changes per hour. Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC. The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

purified water

Controls:

not specified

Amount / concentration applied:

0.5 g (per animal)

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48 and 72 hours

Number of animals:

Number of animals per test: 3 (animals of both sexes were used)

Details on study design:

TEST ITEM PREPARATION
0.5 g (per animal) of FAT 40824/A was weighed as delivered by the Sponsor and then moistened with approximately 0.5 mL of purified water before application.

TREATMENT
Four days before treatment, the left 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 one day before treatment, and regrown fur of all animals was clipped again.
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.
On the day of treatment, 0.5 g of FAT 40824/A was placed on a surgical gauze patch (ca. 4 cm x 4 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.
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.

OBSERVATIONS
Viability/Mortality: Daily from acclimatization of the animals to the termination of test.
Clinical signs: Daily from acclimatization of the animals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation.
SKIN OBSERVATION TIME POINTS AND TERMINATION: The skin reaction was assessed at approximately 1, 24, 48 and 72 hours, as well as 7 and 10 days after exposure (removal of the dressing, gauze patch and test item).
All rabbits were sacrificed by an intravenous injection of Vetanarcol into the ear vein at a dose of at least 1 mL/kg body weight (equivalent to 162 mg sodium pentobarbitone/kg body weight) and discarded.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Remarks:

In animal no 30 erythema was not assessable due to staining produced by the test item

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Signs such as erythema could not be evaluated in all animals at the 1-hour reading due to red marked staining produced by the test item at the application site which persisted in one animal until the 24-hour examination. Where evaluation was permitted, the test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
The test item caused a slight to marked red staining at the test site of all animals from the 1- to the 72-hour reading and persisted as slight staining in one animal until test day 7.
The body weights of all rabbits were considered to be within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40824/A is considered to be "not irritating" to rabbit skin.

Executive summary:

In a GLP compliant study, the primary skin irritation potential of test article was investigated according to OECD test guideline No. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank 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, as well as 7 and 10 days after removal of the dressing.

Signs such as erythema could not be evaluated in all animals at the 1-hour reading due to red marked staining produced by the test item at the application site which persisted in one animal until the 24-hour examination. Where evaluation was permitted, the test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

The application of test article to the skin did however cause a slight to marked red staining at the test site of all animals from the 1- to the 72-hour reading and persisted as slight staining in one animal until test day 7. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed. Based on the study results, FAT 40824/A is considered to be "not irritating" to rabbit 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:

25 January 2006 to 02 March 2006

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:

not specified

Qualifier:

according to guideline

Guideline:

other: Commission Directive 2004/73/EC, B-5, "Acute Toxicity; Eye Irritation/Corrosion", April 29, 2004.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Storage: At room temperature at about 20°C in a desiccator (hygroscopic)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
Test system: Young Adult New Zealand White Rabbit, SPF
Source: Charles River Deutschland GmbH Stolzenseeweg 32-36 D-88353 Kisslegg / German
Age at treatment: 13-14 weeks (male), 12 weeks (females)
Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
Accommodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 57/05, Provimi Kliba AG) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 74/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Results of analysis for contaminants are archived at RCC Ltd.
Water: Community tap water from Füllinsdorf, ad libitum.
Allocation: Male No. 33, Female Nos. 34 and 35

ENVIRONMENTAL CONDITIONS
Air-conditioned with ranges for room temperature 17-23 relative humidity 30-70 % and approximately 10-15 air changes per hour. Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC. The animals were provided with an automatically controlled tight cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g (per animal)

Duration of treatment / exposure:

one second

Observation period (in vivo):

1, 24, 48 and 72 hours after administration

Number of animals or in vitro replicates:

Number of animals per test: 3 (animals of both sexes were used)

Details on study design:

TEST ITEM PREPARATION
0.1 g (per animal) of FAT 40824/A was weighed and applied undiluted as it was delivered by the Sponsor. The eyes of the animals were examined one day prior to test item administration.

TREATMENT
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.
On the day of treatment, 0.1 g of FAT 40824/A was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently hefd together for about one second to prevent loss of test item. The right eye remained untreated and served as the reference control. The treated eyes were not rinsed after instillation.

Observations
viabilily/Mortality and Clinical signs: Daily from acclimatization of the animals to the termination of test.
body weights: At start of acclimatization, on the day of application and at termination of observation
Irritation scores:The eyes of each animal were examined approximately 1, 24, 48, 72 hours and 7, 10, 14, 17 and 21 days after administration.
No necropsy was performed on the animals sacrificed at termination of observation. All rabbits were sacrificed by an intravenous injection of Vetanarcol into the ear vein at a dose of at least 1 mL/kg body weight (equivalent to 162 mg sodium pentobarbitone/kg body weight) and discarded.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animal

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 animal

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0.11

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No abnormal findings were observed in the cornea or iris of any animal at any of the measurement intervals.
Slight reddening of the conjunctivae was noted in alt animals at the 1-hour observation and persisted in one animal until the 24-hour observation.
Slight swelling (chemosis) of the conjunctivae was observed in one animal at the 1-hoar observation.
Marked red staining prevented the assessment of the sclerae in one animal at the 1-hour reading and slight reddening of the sclerae was present in the two remaining animals at the 1-hour observation and persisted in one animal at the 24-hour reading.
No abnormal findings were observed in the treated eye of any animal 43 hours after treatment, the end of the observation period for all animals.
No corrosion of the cornea was observed at any of the reading times.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
Slight to marked red staining of the treated eyes was observed in all animals one hour after treatment and persisted as slight staining until the 72-hour, 7-day or 21-day observation, respectively.
The body weights of all rabbits were considered to be within the normal range of variability.

Individual eye score:

Animal No.	Time point	Corneal opacity	Area of Corneal Opacity	Iris	Conjunctiva		Sclera
					Redness	Chemosis
33	After 24 h	0	0	0	0	0	0
34		0	0	0	1	0	0
35		0	0	0	0	0	1
33	After 48 h	0	0	0	0	0	0
34		0	0	0	0	0	0
35		0	0	0	0	0	0
33	After 72 h	0	0	0	0	0	0
34		0	0	0	0	0	0
35		0	0	0	0	0	0

 

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the study, FAT 40824/A is non-irritant to the eye of rabbits.

Executive summary:

In a GLP compliant study, the primary eye irritation potential of FAT 40824/A was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14, 17 and 21 days after test item instillation.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and Chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00, 0.33 and 0.00 for reddening and 0.00 for Chemosis for all three animals.

The instillation of FAT 40824/A into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae and Chemosis. These effects were reversible and were no longer evident 48 hours after treatment. Slight to marked red staining of the treated eyes was observed in all animals one hour after treatment and persisted as slight staining until the 72-hour, 7-day or 21-day observation, respectively. No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs were observed. Thus, the test item did not induce significant damage to the rabbit eye.

 

Based on the study results, FAT 40824/A does not have to be classified and labelled as irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation study:

In a GLP compliant study, the primary skin irritation potential of test article was investigated according to OECD test guideline No. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank 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, as well as 7 and 10 days after removal of the dressing.

Signs such as erythema could not be evaluated in all animals at the 1-hour reading due to red marked staining produced by the test item at the application site which persisted in one animal until the 24-hour examination. Where evaluation was permitted, the test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

The application of test article to the skin did however cause a slight to marked red staining at the test site of all animals from the 1- to the 72-hour reading and persisted as slight staining in one animal until test day 7. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed. Based on the study results, FAT 40824/A is considered to be "not irritating" to rabbit skin.

Eye irritation study:

In a GLP compliant study, the primary eye irritation potential of FAT 40824/A was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14, 17 and 21 days after test item instillation.

The instillation of FAT 40824/A into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae and Chemosis. These effects were reversible and were no longer evident 48 hours after treatment. Slight to marked red staining of the treated eyes was observed in all animals one hour after treatment and persisted as slight staining until the 72-hour, 7-day or 21-day observation, respectively. No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs were observed. Thus, the test item did not induce significant damage to the rabbit eye.

 

Based on the study results, FAT 40824/A does not have to be classified and labelled as irritant.

Justification for classification or non-classification

Based on the findings in the skin and eye irritation study, the test substance does not considered to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.