Disodium [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]-7-nitronaphthalene-1-sulphonato(3-)]chromate(2-)

Endpoint summary

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:

experimental study

Adequacy of study:

key study

Study period:

Februaryfrom 1st to 4th, 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 24 April 2002

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

semiocclusive

Preparation of test site:

other: intact skin

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount: 0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 rabbits

Details on study design:

Skin reactions were recorded 1, 24, 48 and 72 hours after administration.

SCORING SYSTEM
Primary Irritation Index:
0: non-irritant
> 0 < 2: Mild irritant
> 2 < 5: Moderate irritant
> 5 < 8: Severe irritant

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hrs

Score:

< 2.3

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hrs

Score:

< 2.3

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritant / corrosive response data:

Primary irritation index: 0.0
No evidence of skin irritation was noted.

Individual Skin Reactions Following 4-Hour Exposure Period

Skin Reaction	Observation time	Individual Scores - Rabbit Number and Sex			Total
		35 Male	40 Male	41 Male
Erythema/Eschar formation	1-h	0 STA	0 STA	0 STA	0
	24-h	0 STA	0 STA	0 STA	0
	48-h	0 STA	0 STA	0 STA	0
	72-h	0 STA	0 STA	0 STA	0
Oedema formation	1-h	0	0	0	0
	24-h	0	0	0	0
	48-h	0	0	0	0
	72-h	0	0	0	0
Sum of 24 and 72 Hour Readings (S)					0
Primary Irritation Index (S/6) and Classification*					0/6 = 0.0

Interpretation of results:

not classified

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

Not irritating

Executive summary:

Method

The study was performed to assess the irritation of the test material to the skin of the New Zealand White rabbit. The method followed OECD Guidelines for Testing of Chemicals (24 April 2002) No. 404 "Acute Dermal Irritation/Corrosion" and Method B4 of Commission Directive 2004/73/EC (which constitutes Annex V of Council Directive 67/548/EEC) .

A single 4-hour semi-occluded application (0.5 g) of the test material was administered to the intact skin of three rabbits. Skin reactions were recorded 1, 24, 48 and 72 hours after administration.

Results and conclusion

Primary Irritation Index: 0.0

The mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions.

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:

February from 7th to 17th, 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 24 April 2002

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Amount / concentration applied:

A single application of 0.1 ml (51.3 mg) of the test material was administered to the non irrigated eye of three rabbits.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

Ocular reactions were recorded 1,24,48 and 72 hours after administration.

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hrs

Score:

< 1

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hrs

Score:

< 1

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hrs

Score:

< 2

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hrs

Score:

< 2

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0.66

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0.66

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0.33

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0.66

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0.33

Irritant / corrosive response data:

The test material produced iridial inflammation and minimal to moderate conjunctival irritation. All treated eyes appeared normal at the 72-hour observation.
Maximum Group Mean Score: 12.7. Mild irritant (Class 4 on A 1-8 scale)

Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	97 Male					104 Male					105 Male
	IPR = 2					IPR = 3					IPR = 0+
Time After Treatment	1 hour	24 hours	48 hours	72 hours	Mean 24, 48 and 72 hrs	1 hour	24 hours	48 hours	72 hours	Mean 24, 48 and 72 hrs	1 hour	24 hours	48 hours	72 hours	Mean 24, 48 and 72 hrs
CORNEA
E = Degree of Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
F = Area of Cornea Involved	0	0	0	0	/	0	0	0	0	/	0	0	0	0	/
Score (E x F) x 5	0	0	0	0		0	0	0	0		0	0	0	0
IRITIS
D	0	0	0	0	0	1	0	0	0	0	1	0	0	0	0
Score (D x 5)	0	0	0	0	/	5	0	0	0	/	5	0	0	0	/
CONJUNCTIVAE
A = Redness	1 Sf	1 Sf	1 Sf	0 Sf	0.7	2 Sf	1 Sf	1 Sf	0 Sf	0.7	1 Sf	1 Sf	1 Sf	0 Sf	0.7
B = Chemosis	1	0	0	0	0	2	1	1	0	0.7	2	1	0	0	0.3
C = Discharge	1 Re	1Re	0	0	/	2 Re	1 Re	1 Re	0	/	2	1	1	0	/
Score (A + B + C) x 2	6	4	2	0		12	6	6	0		10	6	4	0
Total Score	6	4	2	0		17	6	6	0		15	6	4	0

IPR = Initial pain reaction

+ = One drop of local anaesthetic instilled into both eyes 1 to 2 minutes before treatment

Sf = Black staining of the fur around the treated eye

Re = Black residual test material

Interpretation of results:

not classified

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

Not irritating.

Executive summary:

Method

The study was performed to assess the irritation of the test material to the eye of the New Zealand White rabbit (SPL Standard Test Method 560.09). The method followed OECD Guidelines for Testing of Chemicals (24 April 2002) No. 405 "Acute Eye Irritation/Corrosion" and Method B5 of Commission Directive 2004/73/EC (which constitutes Annex V of Council Directive 67/548/EEC).

A single application of 0.1 ml (51.3 mg) of the test material was administered to the non irrigated eye of three rabbits. Ocular reactions were recorded 1, 24, 48 and 72 hours after administration.

Results

The test material produced iridial inflammation and minimal to moderate conjunctival irritation. All treated eyes appeared normal at the 72 hour observation.

Maximum Group Mean Score: 12.7

Mild irritant (Class 4 on A 1-8 scale).

Conclusion

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all animals.

Therefore the substance does not meet the criteria to be classified as eye irritating.

In conclusion, the test item can be not classified as irritating, according to the CLP Regulation (EC 1272/2008).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

SKIN IRRITATION

Under the experimental conditions employed Acid Brown 282 (ABr282) did not induce skin irritation when applied to the rabbits skin.

EYE IRRITATION

The eye irritation potential of the ABr282 was assessed in tests conducted at different dye concentrations and ABr282 was judged as non irritating.

Two Huntsman's studies (Huntsman Textile Effects (Germany) GmbH, 1984 and 1974) were disregarded despite they were conducted according to internationally accepted testing procedures, because the substance concentration used was low and one of the impurity compounds of the lots tested is tetrahydro-furfurylalcohol (10.5 % and 11.9 %, respectively), which is classified according to the CLP Regulation, Annex VI, as eye irritant, category 2. Therefore, it is not known if the effects recorded should be attributed to the substance itself or if they are determined by the presence of tetrahydro-furfurylalcohol. Furthermore, another relevant aspect, that may be determinant in the assessment, is that it is not known if the solid substance tested was removed within 1 hours after application; in fact, the test guideline version followed (OECD 405, 1981) in the current test did not envisage the removal of the substance within 1 hours, therefore, it cannot be excluded that the irritation potential determined by the powder form may contributed to the reactions observed. In conclusion, the study has been disregarded.

Justification for selection of skin irritation / corrosion endpoint:

Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Justification for selection of eye irritation endpoint:

Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Justification for classification or non-classification

According to the CLP Regulation (EC 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.

In the key study, the mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in at least 2 of 3 animals for both erythema/eschar and oedema reactions.

According to the CLP Regulation (EC 1272/2008) substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes) if, when applied to the eye of an animal, a substance produces at least in 2 of 3 tested animals, a positive response of corneal opacity ≥ 1 and/or iritis ≥ 1, and/or conjunctival redness ≥ 2 and/or conjunctival oedema (chemosis) ≥ 2, calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all animals in the key study.

In conclusion, the substance is not classified for the eye/skin irritation, according to the CLP Regulation (EC 1272/2008).