Octasodium 2-(6-(4-chloro-6-(3-(N-methyl-N-(4-chloro-6-(3,5-disulfonato-2-naphthylazo)-1-hydroxy-6-naphthylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,5-disulfonato-1-hydroxy-2-naphthylazo)naphthalene-1,5-disulfonate

Endpoint summary

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar 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
- Source: Froxfield farms, UK limited
- Age at study initiation: young and adult
- Weight at study initiation: 2974-3895 g
- Housing: The rabbits were housed individually in aluminium sheet cages (65.2 cm × 68.9 cm × 45.7 cm)
- Acclimation period: 6 d prior to test

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): 25-30
- Photoperiod (hrs dark / hrs light): 12 h light and 12 h dark

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: The test substance was moistened with bi-distilled water before application.

Amount / concentration applied:

0.5 g /animal

Duration of treatment / exposure:

4 h

Observation period:

upto 4 d

Number of animals:

Three females

Details on study design:

TEST SITE
- Area of exposure: left flank of the test animals
- Type of wrap if used: treated area covered with piece of surgical quaze which was secured by two crossed strips of surgical tape. This was covered by piece of impermeable rubber sheet wrapped around the trunk of the animal and secured with adhesive impermeable polyethylene tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): gently cleansed with absorbant cotton swabs soaked in warm water to remove the residual amount of test susbtance.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS: 4 d
(indicate if minutes, hours or days)

SCORING SYSTEM:
- Method of calculation: The Draize scale

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

8

Pathological data:

Animal number		Histopathological finding
		Hyperkeratosis	Parakeratosis/surface debris	Acanthosis	Inflamatory cell infiltration	oedema	Hair folicle loss	Subepithelial fibrosis
4	C	0	0	0	1 F	0	0	0	DHP
4	T	0	0	1 D	2 M-F	0	0	0	DHP
5	C	0	0	1 D	1 M-F	0	0	0	SHP
5	T	0	0	1 D	2 M-F	0	0	0	SHP
6	C	0	0	0	0	0	0	0	SHP
6	T	0	0	1 D	0	0	0	0	SHP

C= control, T=test, DHP=dense hair pattern, SHP=sparse hair pattern, F= Focal, M-F=Multifocal, D=Diffuse 0=No reaction, 1=Minimal reaction, 2=Slight reaction, 3=Moderate reaction, 4=Marked reaction, 5=Sever reaction

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the substance was not irritating to rabbit skin.

Executive summary:

A study was conducted to determine the skin irritation potential of the substance according to a method similar to OECD Guideline 404, in compliance with GLP. The primary skin irritation potential of the test substance was investigated by topical application of 0.5 g to 2.5 cm x 2.5 cm of intact dorsal skin of three young New Zealand White rabbits. The duration of the treatment was 4 h. The scoring of skin reactions was performed at 1, 24, 48, 72 and 96 h after removal of the dressing. The scores of each animal at 24, 48 and 72 h were used in calculating the respective mean values for each type of lesion. The test substance caused red staining of the treated skin and obscured the assessment of erythema. Mean edema scores were calculated for each animal by adding together the oedema score at 1, 2 and 3 d readings and dividing the total by three which is 0.0. Mean erythema score could not be calculated, since the staining obscured the assessment of erythema. There were no other visible signs of irritation seen any animal. Histopathological examination revealed a slight inflammatory reaction in two of the test sites. Under the study conditions, the substance was not irritating to rabbit skin (Robinson, 1994).

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:

1993/94

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.5 (Acute Toxicity: 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
- Source: Froxfield Farms UK limited
- Age at study initiation: young adults
- Weight at study initiation: 3200-4648 g
- Housing: individually in aluminium cages (65.2 cm × 68.9 cm × 45.7 cm)
- Diet (e.g. ad libitum): Standard rabbit diet
- Acclimation period: 6 d prior to main study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 ± 2°C
- Humidity (%): 55 ± 5%
- Air changes (per hr): 25-30
- Photoperiod (hrs dark / hrs light): 12 h light and 12 h dark

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

single dose without washing

Observation period (in vivo):

Animal #1: 21 days
Animal #2: 7 days
Animal #3: 10 days

Number of animals or in vitro replicates:

3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): NA
- Time after start of exposure: NA

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 d

Remarks on result:

not determinable

Remarks:

During the first 10 days after application, corneal effects could not be assessed due to staining of the test material. On day 10, no effects were seen.

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

not determinable

Remarks:

due to staining of the test material assessment of reddening was not possible during the entire study period

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

not determinable

Remarks:

due to staining of the test material assessment of reddening was not possible during the entire study period

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 7 d

Remarks on result:

not determinable

Remarks:

due to staining of the test substance, the iris could not be assessed until day 7, at which time no effects were seen

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

not determinable

Remarks:

due to staining of the test material assessment of reddening was not possible during the entire study period

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 10 d

Irritant / corrosive response data:

Approximately one quarter of the test material was displaced from the eye of two animals immediately after application.
Application of the test material caused slight to moderate initial pain (class 2 to 3 on a scale of 0-5).
Parts of the eye, head and front paws of the animals were stained red by the test material, preventing a full assessment of irritation. During the first 10 days after application, corneal effects could only be fully assessed in one animal, partially assessed in another and in the third animal no assessment was possible. In the animal where full assessment was possible no corneal effects were seen. In the other animal (where corneal assessment was partially obscured by the test material staining) slight to mild corneal opacity was recorded affecting up to half of the cornea which persisted up to and including day 4. No corneal effects were seen after this time.
Iridial effects could not be assessed in all animals approximately 1 hour after dosing.. One day after aplication a ful asesment of the iris could be made in 2 animals, and no effects were seen. In the third animal the iris could not be assessed until day 7, at which time no effects were seen.
Conjunctival redness could not be assessed in any of the animals for the whole of the study. Slight to mild chemosis was seen for up to and including 1 hour, 2 days or 7 days. No chemosis was seen after this time.
Slight to severe discharge was seen in all animals for up to and including 1 hour, 1 day or 7 days. Conjunctival discharge had completely cleared after this time.
Additional observations seen included dried secretions on the eyelids seen in one animal.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the substance was not irritating to rabbit eyes.
This test substance was classified under regulation 67/548/EWG to cause serious eye effects due to the not reversible staining of the conjunctiva. As under regulation (EC) No 1272/2008 only persistent effects that impair vision (persistent corneal opacity, discoloration of the cornea by a dye substance) are relevant for classification, the test substance has not to be classified under CLP.

Executive summary:

A study was conducted to determine the eye irritation potential of the substance in New Zealand White rabbits according to OECD Guideline 405, in compliance with GLP. The test substance was applied neat to the left eye of three female rabbits. Animals were then observed for 21 d. The test eye of each rabbit was stained red by test substance, preventing full assessment of irritation. During the first 10 d after application, corneal effects could only be fully assessed in one animal, partially assessed in another animal and not possible in third animal. Where full assessment was possible no corneal effects seen whereas in other animals mild corneal opacity was recorded which persisted up to Day 4. Iridial effects could not be assessed in all animals immediately after administration. After one day full assessment was done on two animals but no effects were seen. In the third animal assessment was done in 7 d and no effects were observed. Conjunctival redness could not assessed in any of the animal for the whole study. Slight to mild chemosis was seen up to 7 d in one animal. Under the study conditions, the substance was not irritating to rabbit eyes (Robinson, 1994).

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 potential of the substance according to a method similar to OECD Guideline 404, in compliance with GLP. The primary skin irritation potential of the test substance was investigated by topical application of 0.5 g to 2.5 cm x 2.5 cm of intact dorsal skin of three young New Zealand White rabbits. The duration of the treatment was 4 h. The scoring of skin reactions was performed at 1, 24, 48, 72 and 96 h after removal of the dressing. The scores of each animal at 24, 48 and 72 h were used in calculating the respective mean values for each type of lesion. The test substance caused red staining of the treated skin and obscured the assessment of erythema. Mean edema scores were calculated for each animal by adding together the oedema score at 1, 2 and 3 d readings and dividing the total by three which is 0.0. Mean erythema score could not be calculated, since the staining obscured the assessment of erythema. There were no other visible signs of irritation seen any animal. Histopathological examination revealed a slight inflammatory reaction in two of the test sites. Under the study conditions, the substance was not irritating to rabbit skin (Robinson, 1994).

Eye irritation:

A study was conducted to determine the eye irritation potential of the substance in New Zealand White rabbits according to OECD Guideline 405, in compliance with GLP. The test substance was applied neat to the left eye of three female rabbits. Animals were then observed for 21 d. The test eye of each rabbit was stained red by test substance, preventing full assessment of irritation. During the first 10 d after application, corneal effects could only be fully assessed in one animal, partially assessed in another animal and not possible in third animal. Where full assessment was possible no corneal effects seen whereas in other animals mild corneal opacity was recorded which persisted up to Day 4. Iridial effects could not be assessed in all animals immediately after administration. After one day full assessment was done on two animals but no effects were seen. In the third animal assessment was done in 7 d and no effects were observed. Conjunctival redness could not assessed in any of the animal for the whole study. Slight to mild chemosis was seen up to 7 d in one animal. Under the study conditions, the substance was not irritating to rabbit eyes (Robinson, 1994).

Justification for classification or non-classification

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