Trisodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonatonaphthyl]azo]-2,5-dimethylphenyl]azo]benzene-1,4-disulphonate

Endpoint summary

Description of key information

Studies carried out to evaluate skin irritation/Corrosion and eye irritation potency of the test substance following OECD guidelines without GLP has demonstrated that the test substance is non-irritant and non-corrosive to rabbit 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:

1974

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.11 (February 1965)

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

(48 h observation not done; involved a 24 h test substance exposure followed by observations at 24 and 72 h; test substance was patched both on abraded and on intact skin of six animals; local and systemic adverse effects not described)

GLP compliance:

no

Remarks:

Pre-GLP study

Specific details on test material used for the study:

Name: FAT 40075/A
Purity: 74.5 %

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bred on the premises of the testing laboratory
- Age at study initiation: 3 to 4 months
- Weight at study initiation: Males: 2.33 kg; Females: 2.06 kg
- Housing: Caged singly
- Diet: A commercial irradiated diet (Styles-Oxoid), ad libitum
- Water: Sterile filtered water, ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 1 °C
- Humidity: 50-70 %
- Photoperiod: Animals were exposed to artificial light for 10 h daily from 08:00 - 18:00 h

Type of coverage:

occlusive

Preparation of test site:

other: both abraded and intact skin areas lateral to the midline of the back

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL (i.e., 500 mg)
- Concentration (if solution): 66.66 %

Duration of treatment / exposure:

24 h

Observation period:

Up to 72 h (The skin reactions were assessed at 24 and 72 h)

Number of animals:

six animals (four males and two females)

Details on study design:

TEST SITE
- 24 h prior to the dermal application, the backs of the animals were shaved over an area consisting of at least 10% of the total body surface. Two test sites lateral to the mid line of the back were used on each animal. Immediately before the application of the test compound, the right hand site was abraded with the point of a sterile hypodermic needle. The abrasions were sufficiently deep to penetrate the stratum corneum but not to damage the dermis. The left hand site remained intact.
- The test substance was applied to each test site on a 2.5 cm² gauze pad. These were covered with aluminium foil secured with "Sleek" adhesive tape. The test sites were then enclosed by a 6 inch wide "Coban" self adhesive bandage, the edges of which were fixed to the skin by strips of "Sleek" in order to retain the test substance in close contact with the skin.

SCORING SYSTEM: Draize scoring system

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

24 h

Score:

1

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

24 h

Score:

1

Max. score:

4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

24 h

Score:

4

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

24 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #4

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #5

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

24 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

animal #6

Time point:

72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

A slight erythema and a slight to severe oedema was seen in all animals involving both intact and abraded sites 24 h after application of the test substance. By 72 h a slight to mild oedema was seen in the abraded sites of 3/6 animals.

Table:

Erythema Formation					Total
Intact			Abraded
Animal no./sex	24 h	72 h	24 h	72 h
1 (Male)	1	0	1	0
2 (Male)	1	0	1	0
3 (Male)	1	0	1	0
4 (Female)	0	0	1	0
5 (Female)	1	0	1	0
6 (Female)	1	0	1	0
Mean	0.83	0.00	1.00	0.00	1.83

Oedema Formation					Total
Intact			Abraded
No. animal	24 h	72 h	24 h	72 h
1 (Male)	2	0	2	0
2 (Male)	4	0	4	2
3 (Male)	3	0	3	0
4 (Female)	1	0	2	2
5 (Female)	2	0	2	0
6 (Female)	3	0	3	1
Mean	2.50	0.00	2.67	0.83	6.00
				Erythema + Oedema	7.83

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was considered to be non-irritating to skin.

Executive summary:

An in vivo study was conducted to evaluate the skin irritation potential of the test substance (at ca. 74.5 % purity) in rabbits according to the US-FHSA Sect. 191.11 (February 1965) method which is similar to OECD Guideline 404 with few deviations. The test substance was applied under two patches (each on an intact and an abraded site) on the backs of six animals (3/sex). Each test site received 0.75 mL (i.e. 500 mg) of 66.66 % solution in water on a 2.5 cm² gauze pad for 24 h under occlusive conditions. Resulting skin lesions were assessed for erythema and oedema at 24 and 72 h after patch removal. Slight erythema and slight to severe oedema was seen in all animals involving both intact and abraded sites 24 h after application of the test substance. By 72 h, a slight to mild oedema was seen in the abraded sites of 3/6 animals. Skin reactions of intact test sites (by 72 h) returned to normal earlier than abraded. Under the study conditions, the test substance was considered to be non-irritating to 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:

1974

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965)

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

(clinical observations for systemic effects were not done)

GLP compliance:

no

Remarks:

(Pre-GLP study)

Specific details on test material used for the study:

Name: FAT 40075/A
Purity: 74.5 %

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred on the premises of the testing laboratory
- Age at study initiation: 3 to 4 months
- Weight at study initiation: Males: 2.33 kg; Females: 2.06 kg
- Housing: Caged singly
- Diet: A commercial irradiated diet (Styles-Oxoid), ad libitum
- Water: Sterile filtered water, ad libitum
- Acclimation period: One week

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 1 °C
- Humidity: 50-70 %
- Photoperiod: Animals were exposed to artificial light for 10 h daily from 08: 00 - 18:00 h

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated right eye

Amount / concentration applied:

- Amount(s) applied: 100 mg

Duration of treatment / exposure:

Unwashed eye: unspecified (not rinsed after single instillation)
Washed eye: 30 sec

Observation period (in vivo):

1, 6, 24, 48 and 72 h.

Number of animals or in vitro replicates:

Six animals (four males and two females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
After 30 seconds the test substance was, as far as possible, flushed out of the eyes of three rabbits with warm water for 1 min.

SCORING SYSTEM: The primary eye irritation index represents the sum of the mean values for cornea, iris and conjunctivae.

CORNEA:
A Opacity and degree of density (most dense area scored)
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

IRIS:
A Values
Normal 0
Folds above normal, congestion, swelling, circumcornea 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

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 Maximum possible score = 20

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

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:

2

Max. score:

3

Reversibility:

fully reversible within: 10 d

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 6 d

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

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 6 d

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 6 d

Irritation parameter:

cornea opacity score

Basis:

animal #4

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 6 d

Irritation parameter:

iris score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #4

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 10

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

cornea opacity score

Basis:

animal: #3, 5 and 6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Test substance was washed 30 sec post application

Irritation parameter:

iris score

Basis:

animal: #3, 5 and 6

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: Test substance was washed 30 sec post application

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 6 d

Remarks on result:

other: Test substance was washed 30 sec post application

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

Remarks on result:

other: Test substance was washed 30 sec post application

Irritation parameter:

conjunctivae score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 6 d

Remarks on result:

other: Test substance was washed 30 sec post application

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Test substance was washed 30 sec post application

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #6

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 6 d

Remarks on result:

other: Test substance was washed 30 sec post application

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Test substance was washed 30 sec post application

Irritant / corrosive response data:

A moderate conjunctival reaction was seen in all animals 1 h after application of the test substance. All conjunctivae were normal by Day 10. A corneal reaction consisting mainly of small areas of opacity and damage to the corneal epithelium was present from 1 h after application of the test substance in 3/6 animals (all unwashed eyes) and in 1/6 (washed eye) from 6 h. In three animals these changes have resolved by Day 2 but in one case changes were present until Day 6 (unwashed eye).

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was considered to be a non-irritant to rabbit eye.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance (at ca. 74.5 % purity) in rabbits according to U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965) which is equivalent to OECD Guideline 405 with deviations. A single dose of 100 mg of test substance was applied into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The untreated eye served as control. The eyes of three animals were rinsed with warm water for 1 minutes after 30 seconds of test substance instillation. Both with (3/6) and without (3/6) washing, ocular reactions were scored at 1, 6, 24, 48 and 72 h and 6 and 10 d post-instillation. Instillation of the test substance elicited conjunctival redness of a mean score of 2 in 2/3 eyes (unwashed) and had resolved within 10 d after instillation. Unwashed eyes (Day 10) returned to normal later than washed (Day 6). Corneal opacity and chemosis was reversible within 72 h. All the scores for Iritis were zero. Conjuctival redness was reversed within 6 days. Under the study conditions, the test substance was considered to be a non-irritant to rabbit eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin:

An in vivo study was conducted to evaluate the skin irritation potential of the test substance (at ca.74.5 % purity) in rabbits according to the US-FHSA Sect. 191.11 (February 1965) method which is similar to OECD Guideline 404 with few deviations. The test substance was applied under two patches (each on an intact and an abraded site) on the backs of six animals (3/sex). Each test site received 0.75 mL (i.e. 500 mg) of 66.66% solution in water on a 2.5 cm² gauze pad for 24 h under occlusive conditions. Resulting skin lesions were assessed for erythema and oedema at 24 and 72 h after patch removal. Slight erythema and slight to severe oedema was seen in all animals involving both intact and abraded sites 24 h after application of the test substance. By 72 h, a slight to mild oedema was seen in the abraded sites of 3/6 animals. Skin reactions of intact test sites (by 72 h) returned to normal earlier than abraded. Under the study conditions, the test substance was considered to be non-irritating to skin.

Eye:

A study was conducted to assess the eye irritancy potential of the test substance (atca. 74.5 % purity) in rabbits according to U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965) which is equivalent to OECD Guideline 405 with deviations. A single dose of 100 mg of test substance was applied into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The untreated eye served as control. The eyes of three animals were rinsed with warm water for 1 minutes after 30 seconds of test substance instillation. Both with (3/6) and without (3/6) washing, ocular reactions were scored at 1, 6, 24, 48 and 72 h and 6 and 10 d post-instillation. Instillation of the test substance elicited conjunctival redness of a mean score of 2 in 2/3 eyes (unwashed) and had resolved within 10 d after instillation. Unwashed eyes (Day 10) returned to normal later than washed (Day 6). Under the study conditions, the test substance was considered to be non irritant to eye.

Justification for classification or non-classification

Based on the available information, Reactive Orange 035 does not need to be classified for corrosion or irritation as per the Regulation (EC) No. 1272/2008.