2-{4-[4-[4-fluoro-6-(2-(2-vinylsulfonylethoxy)ethylamino)-1,3,5-triazin-2-ylamino]phenylazo]phenylazo}naphthalene-4,6,8-trisulfonate, trisodium salt

Endpoint summary

Description of key information

The test substance was found to be not irritating or corrosive to the skin, but caused irreversible damage to the rabbit 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:

22 May 2001 to 07 June 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Stability of test item: Stable under storage conditions
Stability of test item in bi-distilled water: 24 hours at room temperature
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

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: 10-11 weeks
- Weight at study initiation: Male: 1989 g, Female 2007-2099 g
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with autoclaved wood (RCC Ltd, Füllinsdorf) and haysticks for gnawing.
- Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 90/01) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Haysticks (QS no. 56/01) provided by Eberle Nafag AG, CH-9200 Gossau.
- Water: Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: Under laboratory conditions after health examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3 °C
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12, music during light period.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

electric clipper

Vehicle:

water

Remarks:

Test material was applied moistened in distilled water

Controls:

no

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

10 days

Number of animals:

3 (1 male and 2 female)

Details on study design:

- 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 again clipped. 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 40'800/A was placed on a surgical gauze patch (ca. 2.5 cm x 2.5 cm). This gauze patch was applied to approximately 6 cm2 of 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 so that any reactions (erythema) were clearly visible at that time.
- 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.

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

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:

- Irritation: 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.
- Corrosion: No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

- Viability/mortality/clinical signs: No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
- Coloration: Light red staining was observed at the test site of all animals 1 hour after treatment and persisted in one animal up to the 7-day reading and in the other two animals up to the 72-hour reading.
- Body weights: 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 40800/A was found to be not irritating or corrosive.

Executive summary:

In a GLP-compliant skin irritation study, performed according to OECD guideline 404, the test substance (moistened, 0.5 g) was applied, semi-occlusive, to the skin of three New Zealand White rabbits for 4 hours. After 4 hours, the treated skin was flushed with water and observation were made for an additional 10 days. 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. 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. Light red staining was observed at the test site of all animals 1 hour after treatment and persisted in one animal up to the 7 day reading and in the other two animals up to the 72-hour reading. No corrosive effects were noted on the treated skin of any animal at any measuring interval. Based upon the study results, FAT 40800/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:

19 June 2001 to 16 July 2001

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:

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

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Stability of test item: Stable under storage conditions
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

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: Male: 10-11 weeks, Females: 8-9 weeks
- Weight at study initiation: Male: 2012 g, Female: 1942-2044 g
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood (RCC Ltd., Füllinsdorf) and haysticks for gnawing.
- Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 90/01) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Haysticks (QS no. 56/01) provided by Eberle Nafag AG, CH-9200 Gossau.
- Water : Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: Under laboratory conditions after health examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12, music during light period.

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1g

Duration of treatment / exposure:

21 days

Observation period (in vivo):

up to 21 days

Number of animals or in vitro replicates:

3; (1 male, 2 females)

Details on study design:

- Treatment: The eyes of the animals were examined one day prior to test item 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. On the day of treatment, FAT 40800/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 held 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:
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.

- The primary irritation score was calculated by totaling the mean cumulative scores at 24, 48 and 72 hours and then divided by the number of data points.

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

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0.77

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

positive indication of irritation

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:

- Irritation: Application of the test item to healthy rabbit conjunctivae resulted in a primary irritation score of 0.78. The eye reactions (mean values from 24 to 72 hours) consisted of grade 0.00 corneal opacity, grade 0.00 iris lesions, grade 0.78 redness of the conjunctivae and grade 0.00 chemosis of the conjunctivae. No abnormal findings were observed in the cornea or iris of any animal at any reading. Slight reddening of the conjunctivae was noted in all animals 1 hour after treatment and persisted in two animals up to the 72-hour reading and in the other animal up to the 24-hour reading. Slight swelling of the conjunctivae was also observed in all animals 1 hour after treatment A slight watery discharge was evident in all animals at the 1-hour reading. The sclera could not be assessed in one female at the 1-hour reading due to marked orange staining caused by the test item. Slight reddening was however observed in the other two animals at this reading.
- Corrosion: No corrosion of the cornea was observed at any of the reading times.

Other effects:

- Viability/mortality and clinical signs: No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
- Coloration: Slight to marked orange staining produced by the test item was noted in all animals 1 hour after treatment. Slight staining persisted in the male up to the 72-hour reading, in one female up to the 7-day reading and in the other female up to termination on day 21.
- The body weights of all rabbits were considered to be within the normal range of variability.

Individual eye scores

Animal Number	Time point	Cornea	Iris	Conjunctiva
				Redness	Chemosis
22	24 h	0	0	1	0
23		0	0	1	0
24		0	0	1	0
22	48 h	0	0	1	0
23		0	0	0	0
24		0	0	1	0
22	72 h	0	0	1	0
23		0	0	0	0
24		0	0	1	0

Interpretation of results:

Category 1 (irreversible effects on the eye)

Conclusions:

FAT 40800/A was found to be irritating to rabbit's eyes basis of irreversible light orange staining produced by the test item.

Executive summary:

In a GLP-compliant eye irritation study, performed according to OECD guideline 405, the test substance (0.1g) was placed in the conjunctival sac of the left eye of three New Zealand White rabbits. The eyes were examined for 21 days. 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 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 eye reactions (mean values from 24 to 72 hours) consisted of grade 0.00 corneal opacity, grade 0.00 iris lesions, grade 0.78 redness of the conjunctivae and grade 0.00 Chemosis of the conjunctivae. No abnormal findings were observed in the cornea or iris of any animal at any reading. Slight reddening of the conjunctivae was noted in all animals 1 hour after treatment and persisted in two animals up to the 72-hour reading and in the other animal up to the 24-hour reading. Slight swelling of the conjunctivae was also observed in all animals 1 hour after treatment. A slight watery discharge was evident in all animals at the 1-hour reading. The sclera could not be assessed in one female at the 1-hour reading due to marked orange staining caused by the test item. Slight reddening was however observed in the other two animals at this reading. Slight to marked orange staining produced by the test item was noted in all animals 1 hour after treatment. Slight staining persisted in the male up to the 72-hour reading, in one female up to day 7 and in the other female up to termination on day 21. No corrosion was observed at any of the measuring intervals.In conclusion, FAT 40800/A was found to be irritating to rabbit's eyes basis of irreversible light orange staining produced by the test item.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vivo Skin irritation:

In a GLP-compliant skin irritation study, performed according to OECD guideline 404, the test substance (moistened, 0.5g) was applied, semi-occlusive, to the skin of three New Zealand White rabbits for 4 hours (RCC 2001). After 4 hours, the treated skin was flushed with water and observation were made for an additional 10 days. The test substance showed the erythema as well as the edema score of 0 during the whole observation period. No corrosive effect was evident on the skin. Under the conditions of this experiment, the test substance was found to be not irritating or corrosive.

 

In vivo Eye irritation:

In a GLP-compliant eye irritation study, performed according to OECD guideline 405, the test substance (0.1 g) was placed in the conjunctival sac of the left eye of three New Zealand White rabbits (RCC 2001). The eyes were examined for 21 days. The test substance showed a mean conjunctivae score of 1 at 1 and 24 hours, 0.67 at 48 and 72 hours and 0 at the other time points (7-21 days). The chemosis score was 1 at 1 hour and 0 at the other time points (24 hours – 21 days). Cornea and iris scores were 0 during the whole observation period. The primary irritation score was 0.78. No corrosion of the cornea was observed at any of the measuring intervals. However orange staining of the eye persisted up to and including the end of the observation period. Under the conditions of this experiment, the test substance caused irreversible effects on the eye.

Justification for classification or non-classification

Based on the findings in the skin 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 to be irritating to the skin. However, based on the findings in the eye irritation study, the test substance need to be classified as eye irritant Category 1 according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.