Disodium [4-hydroxy-3-[(2-hydroxy-4-nitrophenyl)azo]naphthalene-1-sulphonato(3-)][1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphtholato(2-)]chromate(2-)

Endpoint summary

Description of key information

Acid Blue 317 is considered as non-irritant on 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:

1994

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)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Test article: FAT 20011/D
Additional specification: Lanacron Marine S-G roh feucht
Batch No.: 61
Purity/Contents: CA. 52.5 %
Physical properties: solid
Storage conditions: room temperature
Validity: July 1999
Safety precautions: gloves and face masks
Test material received: March 22, 1994

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

The test was performed on 3 male rabbits, checked for normal skin conditions, weighing between 2190 to 2310 g. The animals were housed individually in metal cages, identified by ear tattoo, 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.

TEST ANIMALS
- Source: Chemisch-pharmazeutische Fabrik D-88397 Biberach
- Weight at study initiation: between 2190 to 2310 g
- Housing: The animals were housed individually in metal cages, identified by ear tattoo.
- Diet: The rabbits received ad libitum standard rabbit pellet - Nafag No. 814, Gossau, Switzerland - and fresh water.
- Water:ad libitum
- Acclimatisation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Photoperiod: 12 hours light cycle day

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

other: A control gauze patch moistened with distilled water was applied to the contralateral flank.

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

3 days

Number of animals:

3 males

Details on study design:

TEST SITE
An area of at least 36 cm² was shaved on both flanks of the animals approximately 24 hours before treatment. A gauze patch (approx. 12-16 cm²) bearing 0.5 g of the test article was applied to the right flank of each animal. The skin area exposed directly to the test article was approx. 6 cm². A control gauze patch moistened with distilled water was applied to the contralateral flank. The patches were loosely covered with an aluminum foil (approx. 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 (only findings reported).
The skin reactions were evaluated 1, 24, 48, and 72 hours after removing the gauze patches according to the OECD scoring system.

SCORING SYSTEM:
SCORE FOR SKIN IRRITATION IN RABBITS:
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
Total possible erythema score 4

Edema formation
- No edema:..................................................................................................................... 0
- Very slight edema (barely perceptible):............................................................................. 1
- Slight edema (edges of area well defined by definite raising):............................................... 2
- Moderate edema (raised more than 1 mm):........................................................................ 3
- Severe edema (raised more than 1 mm and extending beyond area of exposure:................... 4
Total possible edema score 4

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Other effects:

The body weight growth was not affected by the exposure.

Overall results:

3 animals or less	Mean score animal 1*	Mean score animal 2*	Mean score animal 3*	Max value	Maximum duration of any effect	Max value at the end of the observation period
Erythema/eschar	0.33	0.33	0.33	1	24 hours	0
Edema	0	0	0	1	1 hour	0

* Calculated on the basis of the scores at 24, 48 and 72 hours for each animal.

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 20011/D is not irritating to the rabbit skin.

Executive summary:

The aim of this study was to determine the potential of FAT 20011/D to cause irritation on skin. The study was carried out according to OECD Guideline 404 and EU Method B.4. An area of at least 36 cm² was shaved on both flanks of the three New Zealand rabbits. A gauze patch bearing 0.5 g of the test article was applied to the right flank of each animal. The skin area exposed directly to the test article was approx. 6 cm². A control gauze patch moistened with distilled water was applied to the contralateral flank. The patches were loosely covered with an aluminum foil and held in place for 4 hours by an adhesive tape. The skin reactions were evaluated 1, 24, 48, and 72 hours after removing the gauze patches according to the OECD scoring system. The mean score for erythema after 24/48/72 hrs was found to be 0.33 and the mean edema score was found to be 0. The skin reactions observed were reversible by 48 h observation. As the mean values of the recordings 24 to 72 hours after application were below the threshold of significance (2 score for erythema or edema) FAT 20011/D can be classified as non-irritant.

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:

1994

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

Specific details on test material used for the study:

Test article: FAT20011/D
Additional specification: Lanacron Marine S-Grohfeucht
Batch No.: 61
Purity/Contents: CA.52.5%
Physical properties: Solid
Storage conditions: room temperature
Validity: July 1999
Safety precautions: gloves and face masks
Test material received: March22, 1994

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Chemisch-pharmazeutische Fabrik, D-88397 Biberach.
- Weight at study initiation: 1040-2340 g.
- Housing: Animalss were housed individually in metal cage, identified by ear tattoo.
- Diet : Rabbit food, NAFAG, N°. 814 Tox, NAFAG AG, Gossau, SG (Switzerland); ad libitum.
- Water: e.g. ad libitum.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20+/-3.
- Humidity (%): 30-70.
- Photoperiod: 12 hours light/day.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

One application

Observation period (in vivo):

7 days, because reactions were observed within 72 hours after instillation of the test article.

Number of animals or in vitro replicates:

3 rabbits (1 male and 2 females)

Details on study design:

Application of the substance:
The test article was applied with a spatula. 100 mg of FAT 20011/D was placed into the conjunctival sac of the left 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 article. The right eye remained untreated and served as a control. The animals were checked daily for mortality. The ocular reactions were evaluated 1, 24, 48, and 72 hours after the instillation of FAT 20011/D according to the OECD scoring system.

SCORING SYSTEM: Scale for scoring Ocular Lesions.
Cornea
Opacity-degree of density (area most dense taken for reading)
- No ulceration or opacity: 0
- Scattered or diffuse area 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 area, no details of iris visible, size of pupil barely discernible: 3
- Opaque cornea, iris not discernible through the opacity: 4

Iris
- Normal: 0
- Markedly deepended 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

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

Chemosis
Lids and or nictating membrane
- No swelling: 0
- Any swelling above normal (includes nictitating membrane): 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:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

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:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

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:

0.33

Max. score:

2

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 24h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

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:

FAT 20011/D is considered to be non-irritant to the eyes rabbits.

Executive summary:

The purpose of this study was to determine the eye irritating potency of FAT 20011/D. The study was performed according to OECD Guideline 405 and EU Method B.5, in compliance with GLP. 3 rabbits were used to performed the test (1 males and 2 females). 100 mg of FAT 20011/D was placed into the conjunctival sac of the left 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 article. The right 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, and 72 hours after the instillation of FAT 20011/D according to the OECD scoring system. Mild irritation reactions were seen with iris and conjunctiva, which were mostly reversible within 7 days. Hence, FAT 20011/C is considered to be non-irritant to the eyes rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin

The potential of FAT 20011/D to cause irritation on skin was assessed in a study carried out according to OECD Guideline 404 and EU Method B.4. The mean score for erythema after 24/48/72 h was found to be 0.33 and the mean edema score was found to be 0. The skin reactions observed were reversible by 48 h observation. As the mean values of the recordings 24 to 72 hours after application were below the threshold of significance (2 score for erythema or edema) FAT 20011/D can be classified as non-irritant.

The two supporting studies were carried out with FAT 20011/C and FAT 20011/A also support the conclusion that Acid Blue 317 is not irritating to the skin as per the criteria laid out by the regulation (EC) No. 1272/2008.

Eye:

The purpose of this study was to determine the eye irritating potency of FAT 20011/D. The study was performed according to OECD Guideline 405 and EU Method B.5, in compliance with GLP. 3 rabbits were used to performed the test (1 males and 2 females). 100 mg of FAT 20011/D was placed into the conjunctival sac of the left eye of each animal, after gently pulling away the lower lid from the eyeball. Mild irritation reactions were seen with iris and conjunctiva, which were mostly reversible within 7 days. Hence, FAT 20011/C is considered to be non-irritant to the eye in rabbits.

The conclusion that Acid Blue 317 is not irritating to the eyes was further supported by negative outcome from the eye irritation studies with FAT 20011/C and 20011/A conducted in 1982 and 1974 respectively.

Justification for classification or non-classification

The Acid Blue 317 is considered as non-irritant to skin and eye, hence it does not warrant classification as per the regulation (EC) No. 1272/2008.