Registration Dossier

Administrative data

Description of key information

Not skin irritating

Not eye irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
other: data on Acid Blue 225_constituent 1
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
The experiment was conducted on one of the substance components. It should be noted that the lot tested was characterized by the Acid Blue 225_constituent 1 as main component; however, the impurity profile (which includes also the Acid Blue 225_constituent 2) resulted to be similar to that characterizing the substance under assessment (details are given in the document attached to IUCLID section 13).
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Principles of method if other than guideline:
The procedure adopted was the patch-test 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:
no
Species:
rabbit
Strain:
other: Russian Breed
Details on test animals and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 1.5 to 2 kg.
- Housing: in order to prevent injuries by bites, the rabbits were kept separately in V2A wire cages.
- Diet: food (NAFAG, Gossau SG, rabbit food), ad libitum.
- Water: ad libitum.
Type of coverage:
occlusive
Preparation of test site:
other: intact and scarified
Vehicle:
other: 50 % polyethylene glycol trituration (PEG 400)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.5 g
Duration of treatment / exposure:
24 hours
Observation period:
72 hours
Number of animals:
3 males and 3 females
Details on study design:
TEST SITE
- Area of exposure: the shaven skin on 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.
- Type of wrap if used: the patch was covered with a plastic film 5 x 5 cm, which was fixed to the body with adhesive tape.

OBSERVATION TIME POINTS
24 and 72 hours.

SCORING SYSTEM
The primary irritation index, as the measure of the acute skin irritation provoked by the substance, is defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin.

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 mm) 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) 4
Irritation parameter:
erythema score
Basis:
animal: 3/3
Remarks:
both intact and scarified skin
Time point:
24/48/72 h
Score:
< 2.3
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 3/3
Remarks:
both intact and scarified skin
Time point:
24/48/72 h
Score:
< 2.3
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
The primary irritation index of test item was 0.

Evaluation of the Skin Reactions

Animal No. Sex Reaction 24 hrs after application 72 hrs after application
Intact skin Scarified skin Intact skin Scarified skin
1 M Erythema 0 0 0 0
Oedema 0 0 0 0
2 M Erythema 0 0 0 0
Oedema 0 0 0 0
3 M Erythema 0 0 0 0
Oedema 0 0 0 0
4 F Erythema 0 0 0 0
Oedema 0 0 0 0
5 F Erythema 0 0 0 0
Oedema 0 0 0 0
6 F Erythema 0 0 0 0
Oedema 0 0 0 0
Interpretation of results:
other: not classified, according to the CLP Regulation (EC) No 1272/2008
Conclusions:
Non irritating
Executive summary:

A study was performed to determine the skin irritation potential of test item on Russian breed rabbits according to "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO).

3 males and 3 females rabbits were shaved on the whole back and flanks two days before treatment with an electric clipper. The shaven skin on the left side was slightly scarified immediately before treatment. The test item was applied to each side in quantities of 0.5 g; before application a 50 % polyethylene glycol trituration (PEG 400) was made.

The primary irritation index as the measure of the acute skin irritation provoked by the substance is defined as the average of all the ratings found after 24 and 72 hours on the intact and slightly scarified skin.

The primary irritation index, as the measure of the acute irritation to the skin of rabbits, was found to be 0; therefore test item resulted to be as non-irritant to the skin of rabbits.

Discussion and conclusion

The scoring system criteria used to record the severity of the skin reactions followed in the current test are the same as those reported into the OECD guideline, therefore it is possible to re-evaluate the results in accordance with the CLP Regulation (EC) No 1272/2008.

None of the animals showed any observable response to treatment throughout the 72 hours observation period, in both the cases of intact and scarified skin. Therefore, it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.

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:
other: data on Acid Blue 225_constituent 1
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
The experiment was conducted on one of the substance components. It should be noted that the lot tested was characterized by the Acid Blue 225_constituent 1 as main component; however, the impurity profile (which includes also the Acid Blue 225_constituent 2) resulted to be similar to that characterizing the substance under assessment (details are given in the document attached to IUCLID section 13).
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
The procedure adopted was 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:
no
Species:
rabbit
Strain:
other: Russian Breed
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Housing: rabbits were kept separately in V2A wire cages.
- Diet: food (NAFAG, Gossau SG, rabbit food), ad libitum.
- Water: ad libitum.
- Animal selection: only rabbits showing normal ophthalmic findings were included in the test.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 g
Duration of treatment / exposure:
about 30 seconds in 3 rabbits
Number of animals or in vitro replicates:
3 males and 3 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: the treated eyes of three rabbits were each rinsed with 10 ml lukewarm water.
- Time after start of exposure: about 30 seconds after treatment.

TOOL USED TO ASSESS SCORE: slit-lamp

SCORING SYSTEM
The reactions were appraised with a slit-lamp after 24 hours and after 2, 3, 4 and 7 days on the basis of the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the AFDO.

Cornea
A. Opacity-degree of density (area most dense taken for reading)
No opacity 0
Scattered or diffuse area, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris invisible 4

B. Area of cornea involved
One quarter (or less) but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4
A x B x 5 Total maximum = 80

Iris
A. Values
Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, hemorrhage, gross destruction (any or all of these) 2
A x 5

Conjunctivae
A. Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

B. Chemosis
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 about half closed to completely closed 4

C. Discharge
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, and considerable area around the eye 3
(A + B + C) x 2 Total maximum = 20
Irritation parameter:
cornea opacity score
Remarks:
rinsed eye
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 1
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Remarks:
rinsed eye
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 1
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Remarks:
rinsed eye
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 2
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
chemosis score
Remarks:
rinsed eye
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 2
Reversibility:
fully reversible within: 24 hours
Irritant / corrosive response data:
The irritation index was found to be 5.8 for the cornea, 0 for the iris and 5 for the conjunctivae.

Scores when eyes were not rinsed

Days after application

Rabbit 4, male

Rabbit 5, male

Rabbit 6, male

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

1

20

0

12

15

0

12

15

0

12

2

20

0

10

10

0

10

15

0

10

3

15

0

10

10

0

10

10

0

10

4

5

0

8

10

0

8

10

0

6

7

5

0

6

5

0

8

10

0

6

Scores when eyes were rinsed

Days after application

Rabbit 1, female

Rabbit 2, female

Rabbit 3, female

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

Cornea

Iris

Conjunctiva

1

0

0

6

0

0

0

0

0

6

2

0

0

0

0

0

0

0

0

0

3

0

0

0

0

0

0

0

0

0

4

0

0

0

0

0

0

0

0

0

7

0

0

0

0

0

0

0

0

0

Conclusions:
Non irritating
Executive summary:

A study was performed to determine the eye irritation potential of test item in rabbits. The procedure adopted was 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).

The substance was tested on 3 male and 3 female rabbits. Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula. The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.

The total maximum is 80 for the cornea, 10 for the iris and 20 for the conjunctivae. The irritation index was found to be 5.8 for the cornea, 0 the iris and 5 for the conjunctivae.

Discussion and conclusion

It can be noted that in the study, 3 out of 6 animals did not receive rinsing any time after the instillation of the test substance in eyes. Not rinsing the treated eyes may lead to increased irritation response and does not represent the real time scenarios of eyes of workers getting exposed. Normally, any such incident will lead to rinsing of eyes immediately. Hence, not rinsing of eyes is an elevated testing condition and results normally do not have to be taken into account for hazard assessment.

According to the EC classification of the results obtained, the substance can be considered as non irritant to rabbit's eye.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

There are no specific data about the skin/eye irritation potential of Acid Blue 225, therefore the available information on Acid Blue 225_constituent 1 have been taken into consideration. It should be noted that the lot tested was characterized by the Acid Blue 225_constituent 1 as main component; however, the impurity profile (which includes also the Acid Blue 225_constituent 2) resulted to be similar to that characterizing the substance under assessment. Therefore, the data can be considered as adequate and the approach can be considered as suitable (details are given in the document attached to IUCLID section 13).

SKIN IRRITATION

A study was performed to determine the skin irritation potential of test item on Russian breed rabbits according to "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO). 3 males and 3 females rabbits were shaved; the shaved skin on the left side was slightly scarified immediately before treatment. The test item was applied to each side in quantities of 0.5 g; before application a 50 % polyethylene glycol trituration (PEG 400) was made. The primary irritation index, as the measure of the acute irritation to the skin of rabbits, was found to be 0; therefore test item resulted to be as non-irritant to the skin of rabbits.

The scoring system criteria used to record the severity of the skin reactions followed in the current test are the same as those reported into the OECD guideline, therefore it is possible to re-evaluate the results in accordance with the CLP Regulation (EC) No 1272/2008. None of the animals showed any observable response to treatment throughout the 72 hours observation period, in both the cases of intact and scarified skin. Therefore, it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.

EYE IRRITATION

A study was performed to determine the eye irritation potential of test item in rabbits. The procedure adopted was 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). The substance was tested on 3 male and 3 female rabbits. Amounts of 0.1 g of the test substance were introduced into the conjunctival sac; the treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment. The total maximum is 80 for the cornea, 10 for the iris and 20 for the conjunctivae. The irritation index was found to be 5.8 for the cornea, 0 the iris and 5 for the conjunctivae.

It can be noted that in the study, 3 out of 6 animals did not receive rinsing any time after the instillation of the test substance in eyes. Not rinsing the treated eyes may lead to increased irritation response and does not represent the real time scenarios of eyes of workers getting exposed. Normally, any such incident will lead to rinsing of eyes immediately. Hence, not rinsing of eyes is an elevated testing condition and results normally do not have to be taken into account for hazard assessment.

According to the EC classification of the results obtained, the substance can be considered as non irritant to rabbit's eye.

Justification for classification or non-classification

According to the CLP Regulation (EC) No 1272/2008, 3.2 Skin corrosion/irritation section, skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4

hours.

None of the animals showed any observable response to treatment throughout the 72 hours observation period, in both the cases of intact and scarified skin.

According to the CLP Regulation (EC) No 1272/2008, serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance, which is not fully reversible within 21 days of application. Eye irritation means the production of changes in the eye, which are fully reversible within 21 days of application. The irritation index was found to be 5.8 for the cornea, 0 the iris and 5 for the conjunctivae.

It can be noted that in the study, 3 out of 6 animals did not receive rinsing any time after the instillation of the test substance in eyes. Not rinsing the treated eyes may lead to increased irritation response and does not represent the real time scenarios of eyes of workers getting exposed. Normally, any such incident will lead to rinsing of eyes immediately. Hence, not rinsing of eyes is an elevated testing condition and results normally do not have to be taken into account for hazard assessment. According to the EC classification of the results obtained, the substance can be considered as non irritant to rabbit's eye.

In conclusion, the substance is not classified for the eye/skin irritation, according to the CLP Regulation (EC) No. 1272/2008.