2-[N-(2-cyanoethyl)-4-[(2,6-dichloro-4-nitrophenyl)azo]anilino]ethyl acetate

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:

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Body weight: 2.7 - 2.8 kg
Room temperature and relative humidity: 19-25°C and 40-60%, respectively
Light period: 12 h light/12 h dark

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

yes

Amount / concentration applied:

0.5 g of test substance paste

Duration of treatment / exposure:

4 h, after which the test sites were washed

Observation period:

72 h; the skin was checked for irritation (erythema, eschar, oedema) and corrosion (ulcerations, necrosis, scarring) after 24, 48 and 72 h

Number of animals:

3

Details on study design:

Approximately 24 h before the test, the fur was shaved from the dorso-lateral area of the trunk (6 cm x 6 cm). 500 mg of the pulverized test substance were moistened with water and subsequently applied to a hypoallergenic patch (2.5 x 2.5 cm). Patches were held in place with semi-occlusive dressing for 4 h.

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 signs of erythema, eschar and oedema or ulcerations, necrosis, and scarring were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was not considered to be irritant/corrosive to rabbit skin.

Executive summary:

A study was conducted to determine the skin irritation/corrosion potential of the test substance according to OECD Guideline 404 and EU Method B.4. Female New Zealand White rabbits were exposed for 4 h to 0.5 g of the test substance (paste with water). The test site was shaved and the type of coverage applied was semi-occlusive. After 4 h, the skin was washed with water. The skin sites were checked for irritation (Draize scores - erythema, eschar, oedema) and corrosion (ulcerations, necrosis, scarring) after 24, 48 and 72 h. No signs of skin irritation or corrosion were observed during the study. Under the study conditions, the test substance was not considered to be irritant/corrosive to rabbit skin (Suberg, 1985).

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

From February 02, 1993 to March 09, 1993

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:

yes

Remarks:

for some descriptions and examinations; but it did not affect the validity of the study

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Breeder: Interfauna U.K. Ltd.
Acclimatization period: at least 14 d
Body weight: 3.3-3.8 kg
Room temperature and relative humidity: 21 +/- 1.5°C and 40 - 70%, respectively
Light period: 12 h light/12 h dark
Diet and water: standard diet "Ssniff K4" and tap water, ad libitum

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Remarks:

deionized

Controls:

yes

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 h (After 4 h, the exposed skin was carefully washed with water).

Observation period:

14 d with Draize scoring after 1, 24, 48, 72 h and 7 and 14 d

Number of animals:

3

Details on study design:

Approximately 24 h before the test, the fur was shaved from the dorso-lateral area of the trunk (6 cm x 6 cm). 500 mg of the pulverized test substance were moistened with deionized water and subsequently applied to a hypoallergenic patch. Patches were held in place with semi-occlusive dressing for 4 h. The treated skin area was approx. 6 cm2 in size.

Irritation parameter:

erythema score

Remarks:

due to the orange staining of the treated skin, evaluation of reddening was not possible

Basis:

mean

Time point:

24/48/72 h

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

not determinable because of methodological limitations

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:

Evaluation of the erythema of the skin was not possible during the first 72 h due to the intense coloration by the test substance. Nevertheless, no other inflammatory signs (eschar and oedema formation) became apparent within this period and no irritant effects were observed after 72 h.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was not considered to be irritant/corrosive to rabbit skin.

Executive summary:

A study was conducted to determine the skin irritation/corrosion potential of the test substance according to OECD Guideline 404, in compliance with GLP. Female New-Zealand White rabbits were exposed for 4 h to 500 mg of test substance pulverized on a patch. The test site was shaved and the type of coverage applied was semi-occlusive. After 4 h, the skin was washed with water. The skin sites were checked for irritation (Draize scores - erythema, eschar, oedema) and corrosion (ulcerations, necrosis, scarring) after 1, 24, 48, 72 h and 7 and 14 d. No signs of skin irritation or corrosion were observed during the study. Under the study conditions, the test substance was not considered to be irritant/corrosive to rabbit skin (Krötlinger, 1993).

Reference 3

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

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:

supporting study

Study period:

1985

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)

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Hacking & Churchill Ltd Huntingdon, UK
- Age at study initiation: -
- Weight at study initiation: 3.0 - 3.1 kg
- Housing: single in standard cages
- Diet (ad libitum): Ssniff K 4
- Water (ad libitum): tap water


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 40-60
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: Nov 1985

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 µL finely powdered

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48, 72 h and 7 d

Number of animals or in vitro replicates:

3

Details on study design:

After gently pulling the lower lid away from the eyeball, a volume of 100 µL of the pulverized test substance was placed into the conjunctival sac of one eye of each rabbit. The lids were then gently held together for about one second in order to limit loss of the material. 24 h after instillation of the test substance, the treated eye was rinsed with physiological saline.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all rabbits

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:

all rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

all rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was not considered to be irritant to rabbit eye.

Executive summary:

A study was conducted to determine the eye irritation potential of the test substance according to OECD Guideline 405 and EU Method B.5. Male New-Zealand White rabbits were exposed to the test substance for 24 h. After gently pulling the lower lid away from the eye, a volume of 100 µL of pulverized test substance was placed into the conjunctival sac of one eye of each rabbit. The lids were then gently held together for about one second in order to limit loss of the material. 24 h after instillation of the test substance, the treated eye was rinsed with normal saline. Eye irritation (Draize scoring): signs of cornea (opacity, and area affected), iris (hyperaemia, reaction to light), conjunctivae i.e. conjunctiva of bulbus, lids, and nictitating membrane (erythema, chemosis), and discharge were recorded after 1, 24, 48, 72 h and 7 d. In parallel, the opacity of the aqueous humour was also assessed for potential epithelial damage. Two of the three rabbits exhibited positive indications (scores of 1) of irritation i.e. very slight redness of the conjunctiva after 24 h, which were fully reversible within 48 h. Under the study conditions, the test substance was not considered to be irritating to rabbit eye (Suberg, 1985).

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 02, 1993 to March 09, 1993

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:

yes

Remarks:

for some descriptions and examinations; but it did not affect the validity of the study

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Breeder: Interfauna U.K. Ltd.
Acclimatization period: at least 14 d
Body weight: 3.1-3.5 kg
Room temperature and relative humidity: 21 +/- 1.5°C and 40 - 70%, respectively
Light period: 12 h light/12 h dark
Diet and water: standard diet "Ssniff K4" and tap water, ad libitum

Vehicle:

physiological saline

Controls:

yes

Amount / concentration applied:

approx. 68 mg (in 100 µL)

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48, 72 h and 7 d

Number of animals or in vitro replicates:

3

Details on study design:

After gently pulling the lower lid away from the eyeball, a volume of 100 µL of the pulverized test substance (equivalent to approx. 68 mg) was placed into the conjunctival sac of one eye of each rabbit. The lids were then gently held together for about one second in order to limit loss of the material. 24 h after application of the test substance the treated eye was rinsed with normal saline.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all animals

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:

all animals

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:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

no indication of irritation

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 d

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal: #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was not considered to be irritating to rabbit eye.

Executive summary:

A study was conducted to determine the irritation potential of the test substance according to OECD Guideline 405, in compliance with GLP. Female New Zealand White rabbits were exposed to the test substance for 24 h. After gently pulling the lower lid away from the eye, a volume of 100 µL of pulverized test substance (equivalent to approximately 68 mg) was placed into the conjunctival sac of one eye of each rabbit. The lids were then gently held together for about one second in order to limit loss of the material. 24 h after instillation of the test substance, the treated eye was rinsed with normal saline. Eye irritation (Draize scoring): signs of cornea (opacity, and area affected), iris (hyperaemia, reaction to light), conjunctivae i.e. conjunctiva of bulbus, lids, and nictitating membrane (erythema, chemosis), and discharge were recorded after 1, 24, 48, 72 h and 7 d. In parallel, the opacity of the aqueous humour (McDonald and Shadduck method) was also assessed for potential epithelial damage. The three rabbits exhibited positive indications (scores between 1 and 2) of irritation i.e. slight reactions (redness and swelling) of the mucous membranes, which were fully reversible within 7 d. Under the study conditions, the test substance was not considered to be irritating to rabbit eye (Krötlinger, 1993).

Reference 3

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

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 and EU Method B.4. Female New Zealand White rabbits were exposed for 4 h to 0.5 g of the test substance (paste with water). The test site was shaved and the type of coverage applied was semi-occlusive. After 4 h, the skin was washed with water. The skin sites were checked for irritation (Draize scores - erythema, eschar, oedema) and corrosion (ulcerations, necrosis, scarring) after 24, 48 and 72 h. No signs of skin irritation or corrosion were observed during the study. Under the study conditions, the test substance was not considered to be irritant/corrosive to rabbit skin (Suberg, 1985).

A study was conducted to determine the skin irritation/corrosion potential of the test substance according to OECD Guideline 404, in compliance with GLP. Female New-Zealand White rabbits were exposed for 4 h to 500 mg of test substance pulverized on a patch. The test site was shaved and the type of coverage applied was semi-occlusive. After 4 h, the skin was washed with water. The skin sites were checked for irritation (Draize scores - erythema, eschar, oedema) and corrosion (ulcerations, necrosis, scarring) after 1, 24, 48, 72 h and 7 and 14 d. No signs of skin irritation or corrosion were observed during the study. Under the study conditions, the test substance was not considered to be irritant/corrosive to rabbit skin (Krötlinger, 1993).

Eye irritation:

A study was conducted to determine the irritation potential of the test substance according to OECD Guideline 405, in compliance with GLP. Female New Zealand White rabbits were exposed to the test substance for 24 h. After gently pulling the lower lid away from the eye, a volume of 100 µL of pulverized test substance (equivalent to approximately 68 mg) was placed into the conjunctival sac of one eye of each rabbit. The lids were then gently held together for about one second in order to limit loss of the material. 24 h after instillation of the test substance, the treated eye was rinsed with normal saline. Eye irritation (Draize scoring): signs of cornea (opacity, and area affected), iris (hyperaemia, reaction to light), conjunctivae i.e. conjunctiva of bulbus, lids, and nictitating membrane (erythema, chemosis), and discharge were recorded after 1, 24, 48, 72 h and 7 and 14 d. In parallel, the opacity of the aqueous humour (McDonald and Shadduck method) was also assessed for potential epithelial damage. The three rabbits exhibited positive indications (scores between 1 and 2) of irritation i.e. slight reactions (redness and swelling) of the mucous membranes, which were fully reversible within 7 d. Under the study conditions, the test substance was not considered to be irritating to rabbit eye (Krötlinger, 1993).

A study was conducted to determine the eye irritation potential of the test substance according to OECD Guideline 405 and EU Method B.5. Male New-Zealand White rabbits were exposed to the test substance for 24 h. After gently pulling the lower lid away from the eye, a volume of 100 µL of pulverized test substance was placed into the conjunctival sac of one eye of each rabbit. The lids were then gently held together for about one second in order to limit loss of the material. 24 h after instillation of the test substance, the treated eye was rinsed with normal saline. Eye irritation (Draize scoring): signs of cornea (opacity, and area affected), iris (hyperaemia, reaction to light), conjunctivae i.e. conjunctiva of bulbus, lids, and nictitating membrane (erythema, chemosis), and discharge were recorded after 1, 24, 48, 72 h and 7, 14 and 21 d. In parallel, the opacity of the aqueous humour was also assessed for potential epithelial damage. Two of the three rabbits exhibited positive indications (scores of 1) of irritation i.e. very slight redness of the conjunctiva after 24 h, which were fully reversible within 48 h. Under the study conditions, the test substance was not considered to be irritating to rabbit eye (Suberg, 1985).

Justification for classification or non-classification

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