2,2'-[[5-acetamido-4-[(2-bromo-4,6-dinitrophenyl)azo]-2-methoxyphenyl]imino]diethyl diacetate

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Not conducted under GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Breeder: Kleintier farm Madoerin AG, CH-4414 Fuellinsdorf
Acclimatisation period: 5 days
The test was performed on 3 female rabbits, checked for normal skin conditions, weighing between 2280 to 2510 g (approx. 12-14 weeks old) .
The animals were housed individually in metal cages, identified with individually numbered ear tags, kept at a constant room temperature of 20 ± 3 °C, at a relative humidity of 30-70 % and on a 12 hours light cycle day.
The rabbits received ad libitum standard rabbit pellet - Nafag No. 814, Gossau, Switzerland - and fresh water. All batches of the diet are assayed for nutritive ingredients and contaminant level by the manufacturer. Analytical results are available at the animal supply office.
The quality of the drinking water was according to the specifications of the "Schweizerisches Lebensmittelbuch" (Edition 1972) . Results of the routine chemical examination of water at source (Grundwasserfassung Stein) was conducted periodically by the water authority (Baudepartement des Kantons Aargau, Abteilung Gewaesserschutz) are available to Ciba-Geigy Ltd., as well as the results of inhouse chemical analysis by the analytical laboratories of the Pharmaceuticals Division, Ciba-Geigy Ltd.
The body weight was recorded at start and on day 3 of the test.

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

other: A control gauze patch was applied to the contralateral flank.

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 h

Observation period:

1, 24, 48 and 72h

Number of animals:

3 females

Details on study design:

An area of at least 6 cm2 was shaved on both flanks of the animals approximately 24 h before treatment. A gauze patch (20 cm2) bearing
0.5 g of the test substance was applied to the flank. A control gauze patch was a pplied to the contralateral flank. Both gauze patches were moistened before a pplication with distilled water.
The patches were loosely covered with an aluminum foil (36 cm2) and held in place for 4 hours by an adhesive tape (Isoplast, Isoplast AG,
CH-5200 Brugg).
The animals were checked daily for systemic symptoms and mortality.
The skin reactions were evaluated 1, 24, 48, and 72 hours after removing the gauze patches.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Other effects:

No effect on body weight gain

Interpretation of results:

GHS criteria not met

Conclusions:

The substance was tested for skin irritation potential following OECD 404. Under the experimental conditions the substance does not show any irritation potential

Executive summary:

A study was conducted to determine the skin irritation/corrosion potential of the test substance (62.5% purity) according to OECD Guideline 404. Three female New Zealand White rabbits were exposed under semi-occlusive conditions to a patch (20 cm2) containing 500 mg of moistened test substance placed on a shaved flank. A control gauze patch was applied to the contralateral flank. The patches were loosely covered with an aluminium foil and held in place for 4 h with adhesive tape. The animals were checked daily for systemic toxicity and mortality. Skin reactions were evaluated 1, 24, 48 and 72 h after patch removal. Bodyweight changes were recorded. Mean erythema and edema scores for the 24, 48 and 72 h readings were below 0.3 for all animals. Under the study conditions, no skin irritation was observed and the substance was not considered to be irritating to skin (Huntsman, 1986).

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Three New Zealand white rabbits were supplied by Nottingham University, School of Agriculture, Sutton Bonington, Leicestershire, U.K. At the start of the study the animals weighed 2.49 - 2.78 kg and were approximately twelve to sixteen weeks old. After a minimum acclimatisation period
of five days each animal was given a number unique within the study written on the inner sunface of the ear and on a cage label using a black indelible marker pen.
The animals were individually housed in suspended metal cages. Free access to mains drinking water and food ( Rabbi t Diet, Special Diet Services Limited, Witham, Essex, U.K. ) was allowed throughout the study.

The animal room was maintained at a temperature of 19-23 °C and relative humidity of 50 - 60 %. The rate of air exchange was approximately 15 changes per hour and the Iighting was controlled by a time switch to give 12 hours light and 12 hours darkness.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Amount / concentration applied:

A quantity of 0.5 g of the test material moistened with 0.5 mL of distiIIed water

Duration of treatment / exposure:

4 h

Details on study design:

Approximately twenty-four hours prior to the commencement of the test, each of a group of three rabbits was prepared by closely cìipping the fur from the dorsal/flank areas using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.

The moistened sample was introduced under a 2.5 cm x 2.5 cm gauze patch and placed in position on the shorn skin. The patch was secured in
position over the test material with two lengths of adhesive strapping (SLEEK) in the form of a cross. To prevent the animals from interfering with the patches the trunk of each rabbit was wrapped in an elasticated corset (TUBIGRIP ) and the animals were returned to their cages for the duration of
the exposure period.

Four hours after application the corset and patches were removed from each animal and any residual test material removed by gentle swabbing with cotton wool soaked in distilled water followed by ether. Approximately one hour following removal of the patches , and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale i.e. Draize score.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

No erythema or edema was noted at any test site one hour after removal of the patches. Two of the three test sites continued to show no irritation throughout the study period. Very slight erythema was noted at one treated skin site at the 24-hour observation; no irritation was noted at subsequent
observations. AlI treated skin sites showed faint yelIow staining one to 48 hours after treatment. This did not affect assessment of skin reactions.

Interpretation of results:

GHS criteria not met

Conclusions:

The substance was tested for skin irritation potential following OECD 404. Under the experimental conditions no skin irritation was observed.

Executive summary:

A study was conducted to determine the skin irritation/corrosion potential of the test substance according to OECD Guideline 404. The clipped dorsal flank (2.5 x 2.5 cm) of three New-Zealand white rabbits was exposed to 500 mg test substance (97.4% purity) moistened with 0.5 mL of distilled water under semi-occlusive conditions. Four hours after application, the patches were removed and the treated skin area was carefully cleaned with cotton wool soaked in distilled water followed by ether. Approximately 1 h following removal of the patches, then 24, 48 and 72 h later, the test sites were examined for evidence of primary irritation and scored according to the Draize system. No erythema or edema was noted at any test site 1 h after removal of the patches. Two of the three test sites continued to show no irritation throughout the study period. Very slight erythema was noted at one treated skin site at the 24 h observation; no irritation was noted at subsequent time points. All treated skin sites showed faint yellow staining 1 to 48 h after treatment. This did not affect assessment of skin reactions. Under the study conditions, the test substance was not considered to be irritating to skin (Archroma, 1986).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Three New Zealand white strain rabbits were supplied by Nottingham University, School of Agriculture, Sutton Bonington, Leicestershire, U.K.
At the start of the study the animals weighed 2.37 - 2.66 kg and were approximately twelve to sixteen weeks old.
After a minimum acclimatisation period of five days each animal was given a number unique within the study written on the inner surface of the ear and on a cage label using a black indelible marker pen.
The animals were individually housed in suspended metal cages. Free access to mains drinking water and food (Rabbit Diet, Special Diet Services Limited, l.litham, Essex, U.K. ) was allowed throughout the study.
The animal room was maintaÍned at a temperature of 19 - 22 °C and relative humidity of 50 - 55 %. The rate of air exchange was approximately 15 changes per hour and the Iigthing was controlled by a time switch to give 12 hours light and 12 hours darkness.

Controls:

other: untreated left eye served as control

Amount / concentration applied:

0.1 ml (48 mg)

Number of animals or in vitro replicates:

3 females

Details on study design:

Within 24 hours of commencement of the test both eyes of each rabbit provisionally selected, were exami ed for evidence of ocular irritation or defect using an ophthalmoscope. Animals showing evidence of ocular Ie sions were rejected and replaced.
0n the day of the test each rabbit was held firmly but gently until quiet. A volume of 0.1 ml of the test material which was found to weigh 48 mg was placed into the right eye of each rabbit by gently pulling the lower lid away from the eyeball to form a cup into which the test material was
dropped. The upper and lower eyelids were held together for about one second immediately after application to prevent loss of the test material from the eye. The left eye remained untreated and was used for control purposes.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48h

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Iridial inflammation and a dulling of the normal lustre of the cornea were noted in all treated eyes one hour after treatment. The corneas and iris of all animals appeared normal at subsequent observations.
Conjunctivis identified by redeness and swelling was apparent in all treated eyes at the on-hour observation. The irritation regressed and minimal conjunctivis persisted at the 24 h observation. All treated eyes appeared normal 48 h after treatment.

Interpretation of results:

GHS criteria not met

Conclusions:

The substance was tested for eye irritation following OECD 405. Under the experimental conditions the substance did not show any eye irritation potetial

Executive summary:

A study was conducted to determine the eye irritation potential of the test substance (97.4% purity) according to OECD Guideline 405. Three New-Zealand white rabbits received 48 mg (100 µL) of test substance in the conjunctival sac of one eye. The other eye remained untreated and served as negative control. Effects on iris, cornea and conjunctiva were evaluated after 1, 7, 24 and 48 h. Iridal inflammation and a dulling of the normal lustre of the cornea were noted in all treated eyes 1 h after treatment. The corneas and iris of all animals appeared normal at subsequent observations. Conjunctivitis, identified by redness and swelling, was apparent in all treated eyes at the 1 h observation. The irritation regressed and minimal conjunctivitis persisted at the 24 h time point. All treated eyes appeared normal after 48 h. Under the study conditions, the test substance was therefore not considered to be irritating to rabbit eyes (Archroma, 1986).

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Not conducted under GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Breeder: Kleintierfarm Madoerin AG, CH-4414 Fuellinsdorf
Acclimatisation Period: 5 days
The test was performed on 3 female rabbits, checked for normal skin conditions, weighing between 1910 to 2400 g (approx. 12-14 weeks old) .
The animals were housed individually in metal cages, identified with individually numbered ear tags, kept at a constant room temperature of 20 ± 3 °C, at a relative humidity of 30-70 % and on a 12 hours light cycle day.
The rabbits received ad libitum standard rabbit pellet - Nafag No. 814, Gossau, Switzerland - and fresh water.
All batches of the diet are assayed for nutritive ingredients and contaminant level by the manufacturer. Analytical results are available at
the animal supply office. The quality of the drinking water was according to the specifications of the "Schweizerisches Lebensmittelbuch" (Edition 1972) . Results of the routine chemical examination of water at source (Grundwasserfassung Stein) as conducted periodically by the water authority (Baudepartement des Kantons Aargau, Abteilung Gewaesserschütz) are available to Ciba-Geigy Ltd., as well as the results of inhouse chemical analysis by the analytical laboratories of the Pharmaceuticals Division, Ciba-Geigy Ltd.
The body weight was determined at Day 3 and 7

Vehicle:

water

Remarks:

dispersion

Controls:

other: The left eye was left untreated and served as control

Amount / concentration applied:

0.1 ml

Observation period (in vivo):

1, 24, 48 and 72 h, then 7 days

Number of animals or in vitro replicates:

3 females

Details on study design:

0.1 ml of FAT 40235/A was placed into the conjunctival sac of the right eye of each animal, after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of the test material.
The left eye remained untreated and served as a control.
The animals were checked daily for systemic symptoms and mortality.
The ocular reactions were evaluated 1, 24, 48, 72 hours and 7 days after the instillation of FAT 40235/A according to the OECD scoring system
(Appendix 1) . A slit-lamp was used to facilitate the evaluation.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Remarks:

chemosis

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Interpretation of results:

GHS criteria not met

Conclusions:

The substance was tested for eye irritation following OECD 405. Under the experimental conditions the substance does not show any eye irritative/corrosive potential

Executive summary:

A study was conducted to determine the eye irritation potential of the test substance (62.5% purity) according to OECD Guideline 405. Three New-Zealand white rabbits received 0.1 mL of test substance in the conjunctival sac of the right eye. The left eye remained untreated and served as negative control. After 1, 7, 24, 48 and 72 h, then 7 days, the eyes were evaluated for effects on iris, cornea and conjunctiva according to the OECD scoring system. A slit-lamp was used to facilitate the evaluation. Animals were checked daily for systemic symptoms and mortality. Mean 24, 48 and 72 h corneal opacity and iris scores were equivalent to 0. Conjunctival redness and chemosis occurred, resolving at the latest after 7 days. No mortality was observed and the rabbits gained weight normally. Under the study conditions, the test substance was therefore not considered irritating to rabbit eyes (Huntsman, 1986).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

A study was conducted to determine the skin irritation/corrosion potential of the test substance according to OECD Guideline 404. The clipped dorsal flank (2.5 x 2.5 cm) of three New-Zealand white rabbits was exposed to 500 mg test substance (97.4% purity) moistened with 0.5 mL of distilled water under semi-occlusive conditions. Four hours after application, the patches were removed and the treated skin area was carefully cleaned with cotton wool soaked in distilled water followed by ether. Approximately 1 h following removal of the patches, then 24, 48 and 72 h later, the test sites were examined for evidence of primary irritation and scored according to the Draize system. No erythema or edema was noted at any test site 1 h after removal of the patches. Two of the three test sites continued to show no irritation throughout the study period. Very slight erythema was noted at one treated skin site at the 24 h observation; no irritation was noted at subsequent time points. All treated skin sites showed faint yellow staining 1 to 48 h after treatment. This did not affect assessment of skin reactions. Under the study conditions, the test substance was not considered to be irritating to skin (Archroma, 1986).

Another study was conducted to determine the skin irritation/corrosion potential of the test substance (62.5% purity) according to OECD Guideline 404. Three female New Zealand White rabbits were exposed under semi-occlusive conditions to a patch (20 cm2) containing 500 mg of moistened test substance placed on a shaved flank. A control gauze patch was applied to the contralateral flank. The patches were loosely covered with an aluminium foil and held in place for 4 h with adhesive tape. The animals were checked daily for systemic toxicity and mortality. Skin reactions were evaluated 1, 24, 48 and 72 h after patch removal. Bodyweight changes were recorded. Mean erythema and edema scores for the 24, 48 and 72 h readings were below 0.3 for all animals. Under the study conditions, no skin irritation was observed and the substance was not considered to be irritating to skin (Huntsman, 1986).

The results of the above studies do not trigger a classification for skin irritation according to CLP (EC 1272/2008) criteria.

Eye irritation

A study was conducted to determine the eye irritation potential of the test substance (97.4% purity) according to OECD Guideline 405. Three New-Zealand white rabbits received 48 mg (100 µL) of test substance in the conjunctival sac of one eye. The other eye remained untreated and served as negative control. Effects on iris, cornea and conjunctiva were evaluated after 1, 7, 24 and 48 h. Iridal inflammation and a dulling of the normal lustre of the cornea were noted in all treated eyes 1 h after treatment. The corneas and iris of all animals appeared normal at subsequent observations. Conjunctivitis, identified by redness and swelling, was apparent in all treated eyes at the 1 h observation. The irritation regressed and minimal conjunctivitis persisted at the 24 h time point. All treated eyes appeared normal after 48 h. Under the study conditions, the test substance was therefore not considered to be irritating to rabbit eyes (Archroma, 1986).

A second study was conducted to determine the eye irritation potential of the test substance (62.5% purity) according to OECD Guideline 405. Three New-Zealand white rabbits received 0.1 mL of test substance in the conjunctival sac of the right eye. The left eye remained untreated and served as negative control. After 1, 7, 24, 48 and 72 h, then 7 days, the eyes were evaluated for effects on iris, cornea and conjunctiva according to the OECD scoring system. A slit-lamp was used to facilitate the evaluation. Animals were checked daily for systemic symptoms and mortality. Mean 24, 48 and 72 h corneal opacity and iris scores were equivalent to 0. Conjunctival redness and chemosis occurred, resolving at the latest after 7 days. No mortality was observed and the rabbits gained weight normally. Under the study conditions, the test substance was therefore not considered irritating to rabbit eyes (Huntsman, 1986).

The results of the above studies do not trigger a classification for eye irritation according to CLP (EC 1272/2008) criteria.

Justification for classification or non-classification

Based on the results of in vivo studies, no classification for skin and eye irritation is required for the test substance according to CLP (EC 1272/2008) criteria.