Trisodium 2,4-diamino-3,5-bis-[4-(2-sulfonatoethoxy)sulfonyl)phenylazo]benzenesulfonate

Endpoint summary

Description of key information

Reactive Orange 136 was not irritating to rabbit 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:

Dec. 04, 1996 to Jan. 30, 1997

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)

GLP compliance:

yes (incl. QA statement)

Remarks:

German GLP

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96
- Expiration date of the lot/batch: November 30, 2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature (ca. 20°C) away from light.
- Stability under test conditions: stable at storage conditions
- Stability of the test substance in the solvent/vehicle: Stable for at least 48 h in bidistilled water

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. Karl Thomae GmbH Birkendorferstrasse 65 D - 88397 Biberach / Riss
- Age at study initiation: 15 weeks
- Weight at study initiation: male : 2.5 kg; females: 3.2 kg
- Housing: 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 rabbit maintenance diet (batch 71/96) ad libitum from Kliba Mühlen AG, CH-4303 Kaiseraugst
- Water: Community tap water from Itingen, ad libitum
- Acclimation period: Four days under test conditions after health examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 40-709
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 h artificial fluorescent light/12 h dark

IN-LIFE DATES: December 9-12, 1996

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

electric clipper

Vehicle:

water

Remarks:

bi-distilled water

Controls:

not specified

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 h

Observation period:

72 h.

Viability/Mortality: Daily
Clinical Signs: Daily during the observation period.
Body Weights: At the start of acclimatization, at the start of treatment and at termination of observation.
Irritation Scores: The skin reaction was assessed at approximately 1, 24, 48 and 72 h after the removal of the dressing, gauze patch and test article.

Number of animals:

one male, two females

Details on study design:

The test article was moistened with bi-distilled water before application. Approximately 24 h 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 this test.
On the day of treatment, 0.5 g of the test article was applied to approx. 6 cm2 of the intact skin of the clipped area. It was covered with a 2.5 cm x 2.5 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 h. 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.

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:

FAT 45’176/A showed a primary Irritation score of 0.00 when applied to healthy Intact rabbit skin.
Local signs (mean values from 24 to 72 hours) consisted of grade 0.00 erythema and grade 0.00 edema.
No irreversible 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 test and observation period and no mortality occurred.
Light 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:

FAT45'176/A was not irritating to rabbit skin.

Executive summary:

The primary skin irritation potential of FAT 45176/A was investigated in New Zealand White rabbits according to OECD 404 and EU B.4 guideline.

Approximately 24 h 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 the day of treatment, 0.5 g of the test article was applied to approx. 6 cm²of the intact skin of the clipped area. It was covered with a 2.5 cm x 2.5 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 h. 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.

The scoring of skin reactions was performed 1, 24, 48 and 72 h 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.

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred. However, light 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.

Under the conditions of this experiment, FAT45'176/A was found to cause a primary irritation score of 0.00 (max. 8.0) when applied to healthy intact rabbit skin (Edema and erythema score ‘0’). Therefore, FAT45'176/A was 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:

Dec. 11, 1996 to Jan. 28, 1997

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

GLP compliance:

yes (incl. QA statement)

Remarks:

German GLP

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96
- Expiration date of the lot/batch: November 30, 2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature (ca. 20°C) away from light
- Stability under test conditions: stable at storage conditions

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Dr. Karl Thomae GmbH Birkendorferstrasse 65 D - 88397 Biberach / Riss
- Age at study initiation: 15 weeks
- Weight at study initiation: 3.1 kg; females: 3.0 - 3.2 kg
- Housing: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
- Diet: Pelleted standard Kuba 341 rabbit maintenance diet (batch 71/96) ad libitum from Kuba Mühlen AG, CH-4303 Kaiseraugst
- Water: Community tap water from Itingen, ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 40-70
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 h artificial fluorescent light/12 h dark

IN-LIFE DATES: December 16 - 30, 1996

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

one second

Observation period (in vivo):

14 days.

Viability/Mortality: Daily
Clinical Signs: Daily during the observation period.
Body Weights: At the start of acclimatization, on the day of treatment and at termination of observation.
Irritation Scores: The eyes of each animal were examined for ocular irritation approximately 1, 24, 48 and 72 hours, as well as 7 and 14 days after administration.

Number of animals or in vitro replicates:

one male, two females

Details on study design:

The eyes of the animals were examined for intactness once, one day prior to test article administration. Only those animals with no signs of ocular injury or irritation were used on the test.
At treatment, 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. 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.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission Washington, D.C, 20207 was used for additional control purposes (Washington, 1977).

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

Max. score:

3

Remarks on result:

no 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:

FAT 45'176/A showed a primary irritation score of 0.00 when applied to the conjunctival sac of the rabbit eye.
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 test and observation period, and no mortality occurred.
Short- to middle-term light orange staining of the cornea, conjunctivae and sclera were noted in all animals. Staining was reversible by day 14.
The body weight of the animals was within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

FAT45'176/A was not irritating to the rabbit eye.

Executive summary:

An eye irritation study was performed to assess the possible irritation potential when single doses of FAT 45'176/A were placed in the conjunctival sac of rabbit eyes. The study was carried out according to OECD 405 and EU B.5 guideline.

Test carried out by instillation of 0.1 g into one eye of each of three young adult rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately1, 24, 48 and 72 h as well as 7 and 14 days after test article application.

The test article FAT 45,176/A caused short-term swelling of the conjunctivae in one rabbit and increased lacrimation in all rabbits after one hour only. The test article caused light orange staining of various ocular structures and was reversible after two weeks.

The scores of each animal at the following reading times (24, 48 and 72 h) 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 h and then dividing the resulting total by the number of figures. The primary irritation score was 0.00 (Cornea opacity, Iris, Redness and Chemosis score was ‘0’).

Based on the study results, FAT45'176/A was considered non irritating to the rabbit eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritaion/Corrosion:

The primary skin irritation potential of FAT 45176/A was investigated in New Zealand White rabbits according to OECD 404 and EU B.4 guideline.

Approximately 24 h 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 the day of treatment, 0.5 g of the test article was applied to approx. 6 cm²of the intact skin of the clipped area. It was covered with a 2.5 cm x 2.5 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 h. 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.

The scoring of skin reactions was performed 1, 24, 48 and 72 h 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.

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred. However, light 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.

Under the conditions of this experiment, FAT45'176/A was found to cause a primary irritation score of 0.00 (max. 8.0) when applied to healthy intact rabbit skin (Edema and erythema score ‘0’). Therefore, FAT45'176/A was not irritating to rabbit skin.

Eye Irritation:

An eye irritation study was performed to assess the possible irritation potential when single doses of FAT 45'176/A were placed in the conjunctival sac of rabbit eyes. The study was carried out according to OECD 405 and EU B.5 guideline. Test carried out by instillation of 0.1 g into one eye of each of three young adult rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately1, 24, 48 and 72 h as well as 7 and 14 days after test article application. The test article FAT 45,176/A caused short-term swelling of the conjunctivae in one rabbit and increased lacrimation in all rabbits after one hour only. The test article caused light orange staining of various ocular structures and was reversible after two weeks.

The scores of each animal at the following reading times (24, 48 and 72 h) 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 h and then dividing the resulting total by the number of figures. The primary irritation score was 0.00 (Cornea opacity, Iris, Redness and Chemosis score was ‘0’).

Based on the study results, FAT45'176/A was considered non irritating to the rabbit eye.

Justification for classification or non-classification

Based on the findings in the skin and eye irritation study Reactive Orange 136 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 and eye.