Disodium 5-[[4-[(2-bromo-1-oxoallyl)amino]-2-sulphonatophenyl]azo]-4-hydroxy-6-(methylamino)naphthalene-2-sulphonate

Endpoint summary

Description of key information

Reactive Red 066 was found to be not irritating to the skin as well as eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1974

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

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).

Deviations:

yes

Remarks:

Compared to today's guidelines apprasive and occlusive testing conditions have been used representing elevated testing conditions

GLP compliance:

no

Species:

rabbit

Strain:

other: Russian Breed

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.5 to 2 kgs,
- Housing: kept separately in V2A wire cages
- Diet: (NAFAG, Gossau SG, rabbit food) ad libitum
- Water: ad libitum

Type of coverage:

occlusive

Preparation of test site:

abraded

Vehicle:

other: polyethylene glycol

Remarks:

PEG 400

Controls:

not required

Amount / concentration applied:

Before application a 50 % polyethylene glycol trituration (PEG 400) was made. FAT 40063/A was applied to each side in quantities of 0.5 g.

Duration of treatment / exposure:

24 h

Observation period:

24 and 72 h

Number of animals:

6 rabbits (3 males/3 females)

Details on study design:

TEST SITE
- Area of exposure: left side of the body of rabbit
- Type of wrap if used: Gauze patch and covered with plastic film

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data
- Time after start of exposure: 72 h

Irritation parameter:

erythema score

Remarks:

Intact and scarified skin

Basis:

mean

Remarks:

6 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

scarified skin

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

24 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Scarified skin

Basis:

animal #2

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

scarified skin

Basis:

animal #3

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

24 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

scarified skin

Basis:

animal #4

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: within 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

scarified skin

Basis:

animal #5

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Scarified skin

Basis:

animal #6

Time point:

24 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Remarks:

Intact and scarified skin

Basis:

mean

Remarks:

6 animal

Time point:

48 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

mean

Remarks:

6 animal

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

Scarified skin

Basis:

mean

Remarks:

6 animal

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The primary irritation index of FAT 40063/A was 0.7

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 h on the intact and slightly scarified skin.

Compounds producing combined averages (primary irritation indexes) of 2 or less are only mildly irritating; whereas those with indexes from 2 to 5 are moderate irritants, and those with scores above 6 are considered severe irritants.

 

Individual Animal Score: Erythema

Animal Number	Intact Skin		Scarified skin
	24 h	72 h	24 h	72 h
1	0	0	0	0
2	0	0	0	0
3	0	0	0	0
4	0	0	0	0
5	0	0	0	0
6	0	0	0	0

 

Individual Animal Score: Edema

Animal Number	Intact Skin		Scarified skin
	24 h	72 h	24 h	72 h
1	1	0	2	0
2	0	0	1	0
3	1	0	2	0
4	0	0	1	0
5	1	0	2	0
6	2	0	3	0

 

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40063/A is to be considered as a mild irritant to the skin of rabbits. According to the classification and labelling according to GHS system no classification and labelling is required.

Executive summary:

The study was carried out to determine the primary-irritation index with the test substance (FAT 40063/A) on the skin of the rabbits. 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). Six rabbits (3 males/3 females) of the Russian breed, weighing 1.5 to 2 kgs, 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. 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. FAT 40063/A was applied to each side in quantities of 0.5 g. Before application, a 50 % polyethylene glycol trituration (PEG 400) was made. The gauze patches were removed 24 hours after the application. The reaction of the skin was appraised upon removal and 72 hours after it, on the basis of the evaluation scheme. 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 h on the intact and slightly scarified skin. The primary irritation index was found to be 0.7, therefore, FAT 40063/A is to be considered as a mild irritant to the skin of rabbits.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1974

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

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).

Deviations:

yes

Remarks:

not rinsing treated eyes represents an elevated testing condition compared to today's standard guidelines

GLP compliance:

no

Species:

rabbit

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Russian breed
- Housing: Housed separately in V2A wire cages
- Diet: NAFAG, Gossau SG, rabbit food ad libitum
- Water: ad libitum

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated right eye served as a control

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held open for a few seconds. The right eye served as a control. The treated eyes of three female rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.

Observation period (in vivo):

After exposure the reactions were appraised with a slit-lamp after 24 hours and after 2, 3, 4 and 7 days.

Number of animals or in vitro replicates:

6 rabbits (3 male and 3 female)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water.
- Time after start of exposure: about 30 seconds after treatment.

SCORING SYSTEM: "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the AFDO.

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

cornea opacity score

Remarks:

Rinsed and not rinsed

Basis:

mean

Remarks:

Six rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Remarks:

Rinsed and not rinsed

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Eyes rinsed

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #4

Remarks:

Eyes not rinsed

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #4

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #4

Remarks:

Eyes not rinsed

Time point:

48 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #4

Remarks:

Eyes not rinsed

Time point:

48 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #5

Remarks:

Eyes not rinsed

Time point:

24 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #5

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #5

Remarks:

Eyes not rinsed

Time point:

48 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #5

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #5

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #5

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #6

Remarks:

Eyes not rinsed

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #6

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #6

Remarks:

Eyes not rinsed

Time point:

48 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #6

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #6

Remarks:

Eyes not rinsed

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Remarks:

Eyes not rinsed

Basis:

animal #6

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The irritation index was found to be 1.3 for the conjunctival scores that have been fully reversible within 48 hours. In addition, it has to be mentioned that not rinsing treated eyes represents an elevated testing condition compared to today's standard guidelines. Remaining test material may cause continuous mechanical irritation and therefore altering the response to the chemical.

Other effects:

None observed

The total maximum score is 80 for the cornea, 10 for the iris and 20 for the conjunctivae. The irritation index was found to be 0 for the cornea, 0 for the iris and 1.3 for the conjunctivae.

 

Individual animal score: eyes rinsed

Days after application	Rabbit 1			Rabbit 2			Rabbit 3
	Cornea	Iris	Conjunctivae	Cornea	Iris	Conjunctivae	Cornea	Iris	Conjunctivae
1	0	0	0	0	0	0	0	0	0
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
5	0	0	0	0	0	0	0	0	0
6	0	0	0	0	0	0	0	0	0
7	0	0	0	0	0	0	0	0	0

 

Individual animal score: eyes not rinsed

Days after Application	Rabbit 4			Rabbit 5			Rabbit 6
	Cornea	Iris	Conjunctivae	Cornea	Iris	Conjunctivae	Cornea	Iris	Conjunctivae
1	0	0	8	0	0	12	0	0	6
2	0	0	0	0	0	8	0	0	6
3	0	0	0	0	0	0	0	0	0
4	0	0	0	0	0	0	0	0	0
5	0	0	0	0	0	0	0	0	0
6	0	0	0	0	0	0	0	0	0
7	0	0	0	0	0	0	0	0	0

 

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40063/A is to be considered as a minimal irritant to the eye of rabbits.

Executive summary:

FAT 40063/A was subjected to eye irritation test according to the procedure adopted 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 6 rabbits (3 males/3 females) of the Russian breed. Amounts of 0.1 g of the test substance were introduced into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held open for a few seconds. The right eye served as a control. The treated eyes of three rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment. 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.

The primary-irritation index, which serves as a measure of the acute irritation provoked by the substance, was found to be 0 for the cornea, 0 for the iris and 0.0 for the conjunctivae in case of rinsed eyes. Therefore, FAT 40063/A is to be considered as a non-irritant to the eye of rabbits.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The primary-irritation index of FAT 40063 was determined on the skin of the rabbits. Compounds producing combined averages (primary irritation indexes) of 2 or less are only mildly irritating; whereas those with indices from 2 to 5 are moderate irritants, and those with scores above 6 are considered severe irritants. The primary irritation index as the measure of the acute irritation to the skin of rabbits was found to be 0.7. Therefore, FAT 40063/A is to be considered as a mild irritant to the skin of rabbits.

 

FAT 40063/A was subjected to eye irritation test on 6 rabbits of the Russian breed. The primary-irritation index, which serves as a measure of the acute irritation provoked by the substance, was found to be 0 for the cornea, 0 for the iris and 1.3 for the conjunctivae. Therefore, FAT 40063/A is to be considered as a minimal irritant to the eye of rabbits.

Justification for classification or non-classification

Based on the findings in the skin and irritation studies, the Reactive Red 066 is considered to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 as irritating to the skin as well as eye.