Reaction products of diazotised 2-amino-5-nitrobenzenesulfonic acid coupled with resorcinol, subsequently coupled with diazotized 2-amino-4-nitro-6-sulfo-phenol, chelated with iron, sodium salt

Endpoint summary

Description of key information

Not irritant to the skin.

Not irritant to eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

March, 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Source study has reliablity 2. Details on the read across are attached in section 13.

Principles of method if other than guideline:

Code of Federal Regulations Title 16, Section 1500.41

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Buxted Rabbit Co. Ltd.
- Weight at study initiation: 2.0 kg.
- Housing: individually in grid floor cages in a single room.
- Diet: ad libitum.
- Water: ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3°C
- Photoperiod: natural lighting conditions.

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

Duration of treatment / exposure:

24 h

Observation period:

24, 72 h and 7 d (if any irritation was apparent at the 72 h reading).

Number of animals:

6 males

Details on study design:

TEST SITE
- Area of exposure: 2.5 cm × 2.5 cm
- Type of wrap if used: gauze pads (2.5 cm × 2.5 cm) were applied. The patches were secured by adhesive strapping and the whole trunk further occluded with a stockinette bandage.

SCORING SYSTEM:
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 (inluries in depth)
Oedema formation:
0 no oedema
1 very slight oedeina (bareiy pereeptible)
2 slight oedema (edges of area well-defined by definite raising)
3 moderate oedema (raised approximately 1 mm)
4 severe oedema (raised more than 1 mm and extending beyond the area of exposure)

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #4

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #5

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #6

Time point:

other: 24 - 72 Hours

Score:

0

Max. score:

4

Irritant / corrosive response data:

No signs of irritation were noted on both intact and abraded skin.

Interpretation of results:

GHS criteria not met

Conclusions:

Not irritant to the rabbit skin.

Executive summary:

Method:

Skin irritation potential was tested as described into the Federal Register 1973, Volume 38, No. 187, 1500.41. Test substance was applied on both intact and abraded skin of 6 rabbits for 24 h under occlusive coverage. Signs of irritation were scored at 24 h and 72 h after start of exposure according to Draize criteria.

 

Results:

No signs of irritation were noted.

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: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

March 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Source study has reliablity 2. Details on the read across are attached in section 13.

Principles of method if other than guideline:

Code of Federal Regulations Title 16, Section 1500.42, 1973.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Buxted Rabbit Co. Ltd.
- Housing: individually in grid floor cages in a single room.
- Diet: ad libitum.
- Water: ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 3°C
- Photoperiod: natural lighting conditions.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 100 mg

Duration of treatment / exposure:

No rinse

Observation period (in vivo):

1, 24, 48 and 72 h after instillation

Number of animals or in vitro replicates:

6

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM:
Conjunctivae
(A) Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
0 vessels normal
1 vessels definitely injected above normal
2 more diffuse, deeper crimson red, individual vesseis not easily discernible
3 diffuse beefy red
(B) Chemosis
0 no swelling
1 any swelling above normal (includes nictitating membrane)
2 obvious swelling with partial eversion of lids
3 swelling with lids about half closed
4 swelling with lids about half closed to completely closed

Iris
0 normal
1 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)
2 no reaction to light, haemorrhage, gross destruction (any or all of these)

Cornea opacity
0 no opacity
1 scattered or diffuse area, details of iris clearly visible
2 easily discernible translucent areas, details of iris slightly obscured
3 opalescent areas, no details or iris visible size of pupil barely discernible
4 opaque, iris invisible

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Interpretation of results:

GHS criteria not met

Conclusions:

Not irritant to the rabbit eye.

Executive summary:

Method:

Eye irritation potential was tested as described into the Code of Federal Regulations Title 16, Section 1500.42 1973. Test substance was instilled in one eye of 6 rabbits; the untreated eye served as control. No rinse of eye was done. Signs of irritation were scored at 1, 24, 48 and 72 h after start of exposure according to Draize criteria.

 

Results:

No signs of irritation as cornea opacity, iritis and conjunctivae chemosis were noted. Conjunctivae redness was noted in 2/6 rabbits; effects were slight and fully reversible within 72 h after start of exposure.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Skin Irritation:

According to the CLP Regulation (EC 1272/2008), skin corrosion means the production of irreversible damage to the skin, and skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

 

A substance has to be classified as category 1 (corrosive) if it produces destruction of skin tissue, namely, visible necrosis through the epidermis and into the dermis, in at least 1 tested animal after exposure up to a 4 hour duration.

 

A substance has to be classified as category 2 (irritant) if shows:

(1) mean value of ≥ 2,3 - ≤ 4,0 for erythema/ eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal or, if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions; or

(2) inflammation that persists to the end of the observation period normally 14 days in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; or

(3) in some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above.

In case 6 rabbits are used, a response above the threshold of 2.3 in 4 of 6 rabbits implies a classification in category 2.

 

As no irritation was seen, no classification applied.

 

Eye Irritation:

According to the CLP Regulation (EC 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 to the anterior surface of the eye, which is not fully reversible within 21 days of application .

Eye irritation means the production of changes in the eye following the application of test substance to the anterior surface of the eye, which are fully reversible within 21 days of application.

 

A substance has to be classified as category 1 (irreversible effects on the eye), If, when applied to the eye of an animal, a substance produces:

- at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days; and/ or

- at least in 2 of 3 tested animals, a positive response of:

- corneal opacity ≥ 3 and/or

- iritis > 1,5 calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

 

A substance has to be classified as category 2 (irritating to eye) 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 test material, and which fully reverses within an observation period of 21 days.

In case 6 rabbits are used, a response above the threshold in 4 of 6 rabbits implies a classification in category 2.

 

As irritation effects, in terms of conjunctiva redness were below the threshold of classification and recovered within 72 hours, no classification applied.