Reaction products of 4-amino-3-hydroxy-7-nitronaphthalene-1-sulfonic acid and 2′,4′-dihydroxyacetophenone chelated with iron, sodium salts

Endpoint summary

Description of key information

Not irritant to rabbit skin

Not irritant to rabbit eye

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

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 reliability 1. Details on the read across are available in section 13.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Gaukler, D-6050 Offenbach/Main, FRG
- Weight at study initiation: mean 2.91 kg (males), 2.59 kg (females)
- Housing: individually in stainless steel cages with wire mesh walk floors, floor area: 40 cm × 51 cm
- Diet: Ovator Solikanin 4 mm, Muskator-Werke, D-4000 Duesseldorf 1, FRG (about 130 g per animal per day)
- Water: tap water (about 250 ml per animal per day )
- Acclimation period: at least 8 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 24°C
- Humidity: 30 - 70%
- Photoperiod: 12/12 (hrs dark/hrs light)

Type of coverage:

semiocclusive

Preparation of test site:

other: clipping of the fur at least 15 hours before the beginning of the study

Vehicle:

water

Controls:

other: untreated skin site of same animal

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mm layer suspension (thus about 0.5 g of the test substance)
- Concentration (if solution): 50%

Duration of treatment / exposure:

4 h

Observation period:

8 days

Number of animals:

3 (1 male, 2 females )

Details on study design:

TEST SITE
- Area of exposure: third of the back or flank
- Type of wrap if used: test patch (2.5 × 2.5 cm) secured in position with a porous dressing (4 layers of absorbent gauze + porous bandage)

REMOVAL OF TEST SUBSTANCE
- Washing: Lutrol and Lutrol/water (1:1)
- Time after start of exposure: 4 h

SCORING SYSTEM:
Evaluation of erythema and edema:
0 - none
1 - very slight
2 - well-defined
3 - moderate to severe
4 - severe

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Animal 1: male, 2.91 kg

Animal 2: female, 2.69 kg

Animal 3: female, 2.49 kg

 

Readings:

Readings	Animal	Erythema	Edema
4 h	1	1	0
	2	1	0
	3	1	0
24 h	1	0	0
	2	0	0
	3	1	0
48 h	1	0	0
	2	0	0
	3	0	0
72 h	1	0	0
	2	0	0
	3	0	0
8 d	1	0	0
	2	0	0
	3	0	0
mean 24 - 72 h	1	0.0	0.0
	2	0.0	0.0
	3	0.3	0.0
mean 24 - 72 h		0.1	0.0

Interpretation of results:

GHS criteria not met

Conclusions:

Not irritant to the rabbit skin.

Executive summary:

Method:

Skin irritation potential of the substance was assessed according to OECD guideline 404. Test substance was applied to the skin of 3 rabbits, under semiocclusive coverage for 4 h. Signs of irritation in terms of erythema and oedema were recorded.

Results:

No signs of oedema were noted; slight signs of erythema were seen in 1/3 rabbits, which were reversible within 48 hours.

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

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 reliability 1. Details on the read across are available in section 13.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Gaukler, D-6050 Offenbach/Main, FRG
- Weight at study initiation: mean 2.76 kg (males), 2.87 kg (females)
- Housing: individually in cages made of stainless steel with wire mesh walk floors (floor area 40 cm × 51 cm)
- Diet: Ovator Solikanin 4 mm (Muskator-Werke D-4000 Düsseldorf 1, FRG; about 130 g per animal per day)
- Water: tap water (about 250 ml per animal per day)
- Acclimation period: at least 8 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 24°C
- Humidity: 30 - 70%
- Photoperiod: 12/12 (hrs dark/hrs light)

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 ml bulk volume (about 90 mg) of the ground test substance

Duration of treatment / exposure:

Single application

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

3 (2 females, 1 male)

Details on study design:

SCORING SYSTEM:

Cornea
(A) Opacity-degree of density (area most dense taken for reading)
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 of iris visible, size of pupil barely discernible
4 = Opaque, iris invisible

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

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

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 vessels not easily discernible
3 = Diffuse beefy red

(B) Chemosis
0 = No swelling
1 = Any swelling above normal (includes nictitatinq 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

(C) Discharge
0 = No discharge
1 = Any amount different from normal (does not include small amounts observed in inner canthus of normal animals
2 = Discharge with moistening of the lids and hairs just adjacent to lids
3 = Discharge with moistening of the lids and hairs, and considerable area around the eye

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

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 #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Animal 1: female, 2.95 kg

Animal 2: female, 2.78 kg

Animal 3: male, 2.76 kg

 

Readings	animal	cornea		iris	conjunctiva			symptons
		opacity	area		redness	swelling	discharge
1 h	1	0	0	0	0**	2	2	s
	2	0	0	0*	0**	2	2	s
	3	0	0	0*	0**	2	2	s
24 h	1	0	0	0	2	2	1
	2	0	0	0*	2	2	1	s
	3	0	0	0*	2	2	1	s
48 h	4	0	0	0	1	1	0
	5	0	0	0	1	1	0
	6	0	0	0	1	1	0
72 h	1	0	0	0	1	0	0
	2	0	0	0	1	1	0
	3	0	0	0	1	1	1
8 d	1	0	0	0	0	0	0
	2	0	0	0	0	0	0
	3	0	0	0	0	0	0
mean 24 - 72 h	1	0		0	1.3	1.0
	2	0		0	1.3	1.3
	3	0		0	1.3	1.3
Mean		0		0	1.3	1.2

s: irritation index could not be read because of the staining due to the color of the test substance.

*: if in 1 or 2 animals assessments referring to the same characteristic cannot be made at a particular reading, the mean rounded to the nearest whole number of the values that can be read is inserted instead.

**: if in all animals assessments referring to the same CO characteristic cannot be made at a particular reading, zero is inserted instead.

Interpretation of results:

GHS criteria not met

Conclusions:

Not irritant to the eye of rabbit.

Executive summary:

Method:

Eye irritation potential of test substance was tested according to OECD guideline 405. Test substance was applied to the eye of 3 rabbits. Signs of irritation on iris, cornea, and conjunctivae were scored at 24, 48 and 72 h after application. Observations were continued up to day 8.

Results:

No cornea opacity was noted. Signs on iris could not be seen in 2/3 rabbits at the 24 h observation point due to staining; however, scores were 0 at the following observation times. Conjunctivae redness and chemosis were noted at 24, 48 and 72; a mean score of 1.3 was seen in 3/3 and 2/3 rabbits for redness and chemosis, respectively. Such signs were fully reversible within 8 days. Initial irritation was likely due to mechanical effects of the powder in treated eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No data on target substance was available, thus a read across approach on Similar Substance 01 was used. Details on the read across are available in section 13.

Skin:

The potential of test substance to cause skin irritation was assessed in rabbits according to OECD guideline 404. No irritation response was seen upon a single topical application of 0.5 g of test substance to the intact skin of 3 rabbits for 4 hours under semiocclusive conditions. No signs of oedema were noted; slight signs of erythema were noted, being fully reversible within 48 hours.

Eye:

The potential of test substance to cause eye irritation was assessed in rabbits according to OECD guideline 405. Conjunctival reactions (reddening and swelling) were observable in all animals, with a tendency to decrease in the 24-72 h interval. As the eye was exposed to bulk volume (powder) of test substance, such initial irritation might be caused by mechanical effects. However, such effects were fully reversible within 8 days. No other effects were reported.

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.

No oedema was noted and signs of erythema were slight and reversible, thus 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, limited to conjunctiva, were below the threshold of classification and recovered within 8 days, no classification is applied.