Reaction mass of hydrogen [1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphtholato(2-)][1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) , compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1) and hydrogen bis[1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) , compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1) and hydrogen bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphtholato(2-)]chromate(1-) , compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1)

Endpoint summary

Description of key information

In Vivo Skin Irritation: Under the conditions of the study, a single semi-occlusive application of test material to intact rabbit skin for four hours elicited no dermal irritation.

In Vivo Eye Irritation: Under the conditions of the study, the test material did not cause eye irritation.

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:

02 August 1989 and 05 August 1989

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

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Approximately 11 to 13 weeks of age.
- Weight at study initiation: 2.6 to 3.0 kg
- Housing: The rabbits were individually housed in metal cages with perforated floors.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation: The rabbits selected for the study were all acclimated to the laboratory environment.
- Other: All animals were observed daily for signs of ill health or toxic signs.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Approximately 19 °C.
- Humidity (%): 30-70 % relative humidity.
- Air changes (per hr): Air exchange was maintained at approximately 19 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

Approximately 24 h prior to application of the test material, hair was removed with electric clippers from the dorso-lumbar region of each rabbit exposing an area of skin approximately 10 cm square.

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g of test material.

VEHICLE
- The test material was administered as supplied by the Sponsor.

Duration of treatment / exposure:

4 hours

Observation period:

Examination of the treated skin was made on Day 1 (approximately 30 minutes after removal of the patches) and on Days 2, 3 and 4.

Number of animals:

Two females and one male.

Details on study design:

TEST SITE
- Area of exposure: An area of skin approximately 10 cm square.
- Type of wrap if used: A 0.5 g amount of test material was applied under a 2.5 cm square gauze pad moistened with 0.5 mL distilled water to one intact skin site. Each site was occluded with "Elastoplast" elastic adhesive dressing for the exposure period. The animals were not restrained during the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing: The treatment site was washed using water to remove any residual test substance.
- Time after start of exposure: 4 hours.

OBSERVATION TIME POINTS
Examination of the treated skin was made on Day 1 (i.e. approximately 30 minutes after removal of the patches) and on Days 2, 3 and 4.

SCORING SYSTEM:
- Method of calculation: Draize Irritation Score

Grading and scoring of the dermal reactions were performed using the prescribed numerical scoring system as follows:
ERYTHEMA AND ESCHAR FORMATION:
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

OEDEMA FORMATION:
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

None of the animals showed any response to treatment throughout the observation period.

Interpretation of results:

other: Not classified in accordance with EU criteria.

Conclusions:

Under the conditions of the study, a single semi-occlusive application of test material to intact rabbit skin for four hours elicited no dermal irritation.

Executive summary:

The potential of the test material to cause eye irritation was determined in a GLP study which was conducted in accordance with the standardised guideline OECD 404.

The skin irritancy potential of the test material was investigated in New Zealand white rabbits. Approximately 24 hours prior to application of the test material, hair was removed with electric clippers from the dorso-lumbar region of each rabbit exposing an area of skin approximately 10 cm square. A 0.5 g amount of test material, was applied under a 2.5 cm square gauze pad moistened with 0.5 mL distilled water to one intact skin site on each animal. Each treatment site was occluded with "Elastoplast" elastic adhesive dressing for a four hour period. The animals were not restrained during the exposure period and were returned to their cages. At the end of the exposure period, the semi-occlusive dressing and gauze pad were removed and the treatment site was washed using water to remove any residual test material. Examination of the treated skin was made on Day 1 (i.e. approximately 30 minutes after removal of the patches) and on Days 2, 3 and 4.

Under the conditions of the study, a single semi-occlusive application of test material to intact rabbit skin for four hours elicited no dermal irritation.

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:

7 November 1989

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

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: Approximately 11 to 15 weeks of age.
- Weight at study initiation: 2.6 to 3.4 kg
- Housing: The rabbits were individually housed in metal cages with perforated floors.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation: The rabbits selected for the study were all acclimated to the laboratory environment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Approximately 19 °C
- Humidity (%): 30 - 70 % relative humidity.
- Air changes (per hr): Approximately 19 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours dark/ 12 hours light.


.

Vehicle:

unchanged (no vehicle)

Controls:

other: The contralateral eye remained untreated and served as a control.

Amount / concentration applied:

TEST MATERIAL
- Amount applied: Approximately 70 mg of test material, the weight occupying a volume of 0.1 mL; the eyelids were then held gently together for one second before releasing.

Duration of treatment / exposure:

The test material was not removed and therefore the duration of treatment was 7 days.

Observation period (in vivo):

Examination of the eyes was made after 1 hour and 1, 2, 3, 4 and 7 days after instillation. All animals were observed daily for signs of ill health or toxic signs.

Number of animals or in vitro replicates:

Three animals; two males and one female.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: The test material was not removed.

TOOL USED TO ASSESS SCORE: Observation of the eyes was aided by the use of a handheld torch.

SCORING SYSTEM: Grading and scoring of the ocular lesions were performed using the prescribed numerical scoring system as follows:

CORNEA
Opacity: Degree of density (area most dense taken for reading)
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
Easily discernible translucent areas, details of iris slightly obscured: 2
Nacrous areas, no details of iris visible, size of pupil barely discernible: 3
Opaque cornea, iris not discernible through the opacity: 4

AREA OF CORNEA INVOLVED
One quarter (or less) but not zero: 1
Greater than one quarter, but less less half: 2
Greater than half, but less than three quarters: 3
Greater than three quarters, up to whole area: 4

IRIS
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive):1
No reaction to light, haemorrhage, gross destruction (any or all of these): 2

CONJUCTIVAE
Redness (refers to palpebral and bulbar conjunctivae cornea and iris)
Blood vessels normal: 0
Some blood vessels definitely hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse, beefy red: 3

CHEMOSIS (lids and/or nictitating membranes)
No swelling: 0
Any swelling above normal (includes nictitating membranes): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half-closed: 3
Swelling with lids more than half-closed: 4

Irritation parameter:

cornea opacity score

Basis:

other: All three animals tested

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

other: All three animals tested

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

other: All three animals tested

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

other: All three animals tested

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No corneal damage or iridial inflammation was seen throughout the observation period.
Mild conjunctival inflammation was present one hour after instillation only in all three animals.

Interpretation of results:

other: Not classified in accordance with EU criteria.

Conclusions:

Under the conditions of the study, the test material does not cause eye irritation.

Executive summary:

The potential of the test material to cause eye irritation was determined in a GLP study which was conducted in accordance with the standardised guidelines OECD 405.

The eye irritation potential of the test material was investigated in New Zealand White rabbits. The eyes of each animal were examined prior to instillation of the test material to ensure that there was no pre-existing corneal damage or conjunctival inflammation. Approximately 70 mg of test material, the weight occupying a volume of 0.1 mL, was placed into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The contralateral eye remained untreated and served as a control. Examination of the eyes was made after 1 hour and 1, 2, 3, 4 and 7 days after instillation. Observation of the eyes was aided by the use of a handheld torch. The eyes were then graded and scored to determine eye irritation.

Under the conditions of the study, the test material does not cause eye irritation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In Vivo Skin Irritation

The potential of the test material to cause skin irritation was determined in a GLP study which was conducted in accordance with the standardised guideline OECD 404. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

The skin irritancy potential of the test material was investigated in New Zealand white rabbits. Approximately 24 hours prior to application of the test material, hair was removed with electric clippers from the dorso-lumbar region of each rabbit exposing an area of skin approximately 10 cm square. A 0.5 g amount of test material, was applied under a 2.5 cm square gauze pad moistened with 0.5 mL distilled water to one intact skin site on each animal. Each treatment site was occluded with "Elastoplast" elastic adhesive dressing for a four hour period. The animals were not restrained during the exposure period and were returned to their cages. At the end of the exposure period, the semi-occlusive dressing and gauze pad were removed and the treatment site was washed using water to remove any residual test material. Examination of the treated skin was made on Day 1 (i.e. approximately 30 minutes after removal of the patches) and on Days 2, 3 and 4.

Under the conditions of the study, a single semi-occlusive application of test material to intact rabbit skin for four hours elicited no dermal irritation.

In Vivo Eye Irritation

The potential of the test material to cause eye irritation was determined in a GLP study which was conducted in accordance with the standardised guidelines OECD 405. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

The eye irritation potential of the test material was investigated in New Zealand White rabbits. The eyes of each animal were examined prior to instillation of the test material to ensure that there was no pre-existing corneal damage or conjunctival inflammation. Approximately 70 mg of test material, the weight occupying a volume of 0.1 mL, was placed into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The contralateral eye remained untreated and served as a control. Examination of the eyes was made after 1 hour and 1, 2, 3, 4 and 7 days after instillation. Observation of the eyes was aided by the use of a handheld torch. The eyes were then graded and scored to determine eye irritation.

Under the conditions of the study, the test material does not cause eye irritation.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to skin or eye irritation.