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Description of key information

Acid Yellow 220 was slightly irritating to the rabbit, but scores were insufficient to warrant classification. However, it was considered to be irritating to 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:
1974
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
(no untreated skin areas to serve as the control; exposure period is of 24 h; smaller area of application (i.e., approx. 2.5 cm2 gauze pad). As test was performed on scarified skin it is considered as deviation.
Qualifier:
according to guideline
Guideline:
other: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO)
GLP compliance:
no
Specific details on test material used for the study:
Test Item: FAT 20049/A
Purity: approx. 89 %
Species:
rabbit
Strain:
other: Russian
Details on test animals or test system and environmental conditions:
- Weight at study initiation: 1.5 to 2 kg
- Housing: The animals were housed individually in V2A wire cages.
- Diet: Standard rabbit food - NAFAG, Gossau SG, rabbit food (ad libitum)
- Water: ad libitum
Type of coverage:
occlusive
Preparation of test site:
other: shaved and scarified
Vehicle:
other: other: polyethylene glycol (PEG 400)
Controls:
not specified
Amount / concentration applied:
- Amount(s) applied (volume or weight with unit): 500 mg
- Concentration (if solution): Before application a 50 % Polyethylene glycol trituration (PEG 400) was made.
Duration of treatment / exposure:
24 h

Observation period:
72 h
Number of animals:
6 (3 males and 3 females)
Details on study design:
PROCEDURE:
- 2 days prior to the dermal application, six rabbits were shaved on the whole back and flanks with an electric clipper. The shaven skin of the left side was slightly scarified immediately before treatment.
- A gauze patch 2.5 x 2.5 cm was soaked with the test substance and immediately applied to the prepared skin. The patch was covered with a plastic film 5 x 5 cm, which was fixed to the body with adhesive tape.
- Test substance was applied to each side in quantities of 500 mg. Before application a 50 % polyethylene glycol trituration (PEG 400) was made. The gauze patches were removed 24 h after the application. The reaction of the skin was appraised upon removal and 72 h after it.

SCORING SYSTEM:
Dermal irritation was graded in each animal according to the following scoring scale:
- Erythema and eschar formation:
0 no erythema
1 very slight erythema (barely perceptible)
2 well-defined erythema
3 moderate to severe erythema
4 severe erythema (beet redness) to slight eschar formation (injuries in depth)

- Edema formation:
0 no edema
1 very slight edema (barely perceptible)
2 slight edema (edges of area well-defined by definite raising)
3 moderate edema (raised approximately 1 millimeter)
4 severe edema (raised more than 1 millimeter and extending beyond area of exposure)
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: intact skin
Irritant / corrosive response data:
The primary irritation index as the measure of the acute irritation to the skin of rabbits was found to be 0.3
Other effects:
none

Evaluation of the skin reactions:

 

Animal No. (sex)

Skin Reaction

24 h after application

72 h after application

Intact

skin

Scarified

skin

Intact

skin

Scarified

skin

1 (M)

Erythema

0

0

0

0

Edema

0

0

0

0

2 (M)

Erythema

0

0

0

0

Edema

0

2

0

0

3 (M)

Erythema

0

0

0

0

Edema

0

2

0

0

4 (F)

Erythema

0

0

0

0

Edema

0

1

0

0

5 (F)

Erythema

0

0

0

0

Edema

0

1

0

0

6 (F)

Erythema

0

0

0

0

Edema

0

2

0

0

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance was considered to be not irritating to the intact skin.
Executive summary:

An in vivo study was conducted to evaluate the skin irritation potential of the test substance in Russian rabbits according to the method given in the Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO) which is similar to OECD Guideline 404 with few deviations.

In this study, 500 mg of test substance (i.e. 50 % polyethylene glycol trituration (PEG 400)) was applied to each side. A gauze patch of 2.5 x 2.5 cm was soaked with the test substance and immediately applied to the scarified as well as intact skin of rabbits for 24 h under occlusive conditions. After the 24 h of application period, the dressing was removed and the intact and abraded sites were assessed for oedema and erythema. A further assessment was made at 72 h.

No skin reaction was observed the exposed intact skin. Even no erytehma was seen on the scarified skin as well. However, very slight to slight edema was observed on the sacrificed skin of 5 out of 6 animals, which was fully reversible within 72 h. However, exposure to scarified skin is an elevated testing condition, which leads to increased irritation and is a deviation from guideline. Hence the results obtained with scarified skin normally do not have to be taken into account for hazard assessment. Hence, it was concluded that, under the study conditions, the test substance was not irritating to the intact skin.

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:
no data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
(observation period was up to 7 d instead of 21 d; eyes were not examined at 1 h after test substance application; clinical observations for systemic effects was not done)
Qualifier:
according to guideline
Guideline:
other: the technique described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO).
GLP compliance:
not specified
Specific details on test material used for the study:
Test Item: FAT 20049/A
Purity: approx. 89 %
Species:
rabbit
Strain:
other: Russian breed
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Housing: The rabbits were caged singly in V2A wire cages
- Diet: NAFAG, Gossau SG, rabbit food (ad libitum)
- Water: ad libitum
- Only rabbits showing normal ophthalmic findings were included in the test.
Vehicle:
unchanged (no vehicle)
Controls:
other: the right eye served as control
Amount / concentration applied:
- Amount applied: 100 mg
- Concentration: Undiluted
Duration of treatment / exposure:
The treated eyes of three (out of six) rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.
Observation period (in vivo):
7 d

Number of animals or in vitro replicates:
6 (i.e., 3 males and 3 females)
Details on study design:
- The test substance in an amount of 100 mg was inserted into the conjunctival sac of the left eye of the rabbits and the eyelids were held open for a few seconds. The right eye was not treated and served as an untreated control. In 3 of the 6 rabbits approx. 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water.

- The eye irritation was evaluated with a slit-lamp on 1, 2, 3, 4 and 7 d (i.e., 24, 48, 72, 96 and 168 h).

SCORING SYSTEM:
- The ocular reactions were scored by the method described in Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the AFDO.
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
1.9
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Fully reversible within 4 d when rinsed after 30 seconds
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
1.7
Max. score:
3
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: Fully reversible within 7 d when rinsed after 30 seconds
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
2.1
Max. score:
4
Reversibility:
not fully reversible within: 7 d
Remarks on result:
other: Fully reversible within 7 d when rinsed after 30 seconds
Irritant / corrosive response data:
- The irritation index was found to be 15.3 for the cornea, 0 for the iris and 9 for the conjunctivae.
- The observed cornea and conjunctivae reactions in the unwashed eyes were not reversible within the observation period of 7 d.
Other effects:
none

Results of the eye irritation:

Eyes washed after application of test substance:

Days

after

application

Rabbit 1 (F)

Rabbit 2 (F)

Rabbit 3 (F)

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

1

0

0

8

15

0

10

10

0

12

2

0

0

6

5

0

6

10

0

8

3

0

0

6

0

0

0

5

0

6

4

0

0

0

0

0

0

0

0

6

7

0

0

0

0

0

0

0

0

0

 

Unwashed eyes after application of test substance:

 

Days

after

application

Rabbit 4 (M)

Rabbit 5 (M)

Rabbit 6 (M)

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

1

30

0

16

20

0

16

20

0

16

2

30

0

14

30

0

14

20

0

14

3

40

0

14

30

0

12

30

0

14

4

40

0

12

30

0

12

30

0

12

7

40

0

12

5

0

12

20

0

12

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The test substance was considered to be irritating to the eyes of rabbits.

Executive summary:

An in vivo study was conducted to evaluate the acute eye irritation potential of the test substance (at ca. 89 % purity) in rabbits according to the procedure described in the Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics (1959) of the US Association of Food and Drug Officials (AFDO) which is similar to the OECD Guideline 405 but with few deviations.

 

100 mg of the test substance was placed into the conjunctival sac of the left eye of six rabbits (i.e. 3 male and 3 females). The right eye remained untreated and was used as control. In three rabbits, washing was done with 10 mL of lukewarm water after 30 seconds of test substance instillation. Assessment of ocular damage/irritation was made with a slit-lamp at 24, 48, 72, 96 and 168 h following treatment. The irritation index was calculated to be 15.3, 0 and 9 for the cornea, iris and conjunctivae, respectively. The mean scores for corneal opacity, iris, conjunctival redness and chemosis were considered to be 1.9, 0, 1.7 and 2.1. The score obtained for corneal opacity thus fulfils the criteria for classification as eye irritant category 2 as specified by the Regulation (EC) No. 1272/2008. Thus, the test substance was considered irritating to the eyes of rabbits.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

In the in vivo study considered to be key, the skin irritation potential of FAT 20049/A (purity: approx. 89 %) was evaluated according to the method given in the “Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO) which is similar to OECD Guideline 404 with few deviations. In this study, 500 mg of test substance (i.e. 50 % polyethylene glycol trituration (PEG 400)) was applied to the scarified as well as intact skin of rabbits for 24 h under occlusive conditions. No skin reaction was observed the exposed intact skin. Hence, it was concluded that, the test substance was not irritating to the intact skin.

In the vivo study (1984), considered as supporting, conducted according to the OECD Guideline 404, 0.5 ml of FAT 20049/D (purity: approx. 22.5 %) was applied on a 6 cm² gauze pad which was then applied to the intact skin of rabbits for 4 h under occlusive conditions. After the removal of the dressings, the skin reactions were assessed up to 72 h. Due to intensive staining by the test substance a scoring of the erythema values was not possible. The dermal irritation was evaluated after the score for oedema formation only. The mean oedema score was found to be 0 at 24, 48 and 72 h. Hence, it was concluded that, FAT 20049/D was found to be not irritating when applied to the rabbit skin.

In another supporting study (1987) conducted according to OECD Guideline 404, 0.5 ml FAT 20049/E (purity: approx. 27.5 %) was applied on shaved flanks (6 cm2) with a gauze patch (20 cm2) which was then loosely covered with an aluminium foil (36 cm2) for 4 h under occlusive conditions. No skin reactions were seen throughout the observation period. Hence, it was concluded that, FAT 20049/E was not irritating when applied to the rabbit skin.

Eye irritation:

An in vivo study (1974), considered to be key, was conducted to evaluate the acute eye irritation potential of FAT 20049/A (at ca. 89% purity) in rabbits according to the procedure described in the Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics (1959) of the US Association of Food and Drug Officials (AFDO) which is similar to the OECD Guideline 405 but with few deviations. 100 mg of the test substance was placed into the conjunctival sac of the left eye of six rabbits (i.e. 3 males and 3 females). The mean scores for corneal opacity, iris, conjunctival redness and chemosis were considered to be 1.9, 0, 1.7 and 2.1. The score obtained for corneal opacity thus fulfils the criteria for classification as eye irritant category 2 as specified by the Regulation (EC) No. 1272/2008. Thus, the test substance was considered irritating to the eyes of rabbits.

An in vivo study was conducted to evaluate the acute eye irritation potential of FAT 20049/D (27.5 % purity) in rabbits according to OECD Guideline 405. In all the animals the eye reactions were reversed by 21 days. The mean scores obtained for corneal opacity, iris, conjunctival redness and chemosis were 1, 0.28, 2 and 1.4 respectively. Hence, the substance was considered to have met the classification criteria as set by the Regulation (EC) No. 1272/2008. Hence, the test substance was considered irritating to the eyes.

An in vivo study was conducted to evaluate the acute eye irritation potential of FAT 20049/D (27.5 % purity) in rabbits according to OECD Guideline 405. The mean scores obtained for corneal opacity, iris, conjunctival redness and chemosis were 1.89. 0.4, 2.78 and 2.89 respectively. Due to extreme irritation of the conjuctiva in the eyes the animals were killed after 72 hours scoring out of animal protection reasons. Hence, under the conditions of the study, the test substance was considered irritating to the eye.

Justification for classification or non-classification

Acid Yellow 220 was found be not irritating to the skin, but needs to be classified as eye irritant category 2 as per the criteria of Regulation (EC) No. 1272/2008.