Acetic acid, oxo-, sodium salt, reaction products with ethylenediamine and hydroxybenzenesulfonic acid monosodium salt, iron sodium salts

Endpoint summary

Description of key information

In the key skin irritation study conducted with the nearest analogue Fe(3K)EDDHSA (EC No. 462 -490 -6) the average scores calculated for each of the three animals for three consecutive days (24, 48 and 72 hours) do not exceed cut-off-values defined in the CLP. Application of the other closely related substance Fe(Na)EDDHA (CAS 84539-55-9) to the rabbit skin was evaluated as not irritating in two skin irritation studies in albino rabbits.
In the key eye irritation study conducted with Fe(3K)EDDHSA the mean values of the scores calculated for each animal over 24, 48 and 72 hours were 0.7, 0.3 and 0.0 for chemosis, 0.0, 0.0 and 0.0 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity. The values did not exceed the trigger values leading to classification and labelling according to CLP. Two eye irritation studies, performed according to OECD Guideline 405 with the test item Fe(Na)EDDHA (CAS 84539-55-9) in rabbits, showed that the test substance does not cause eye irritation as none of the animals achieved a mean score leading to classification according to CLP.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Study period:

2000-08-08 to 2000-08-13

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EC Directive No. 92/69/EEC, B.4, 31st July 1992.

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France
- Age at study initiation: not reported
- Weight at study initiation: mean body weight of 3 animals 2.6 ± 0.2 kg
- Housing: individually in polystyrene cages (48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.
- Diet (e.g. ad libitum): ad libitum. 110 pelleted diet (UAR, 91360 Villemoisson-sur-Orge, France).
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ±3
- Humidity (%): 30 to 70
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped using electric clippers

Vehicle:

unchanged (no vehicle)

Remarks:

The untreated skin served as control

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg of the test substance were placed on a moistened gauze pad. Purified water (prepared at CIT by reverse osmosis) was used in order to moisten the test substance and ensure a good contact with the skin.

Duration of treatment / exposure:

3 min and 4 hours

Observation period:

The skin was examined approximately 1 hour, 24, 48 and 72 hours after removal of the dressing.

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: back
- % coverage: 6%
- Type of wrap if used: The test substance and the gauze pad were held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residual test substance was wiped off by means of a moistened gauze pad.
- Time after start of exposure: 3 min and 4 hours

SCORING SYSTEM:
Erythema and eschar formation:
- no erythema: 0
- very slight erythema (barely perceptible): 1
- well-defined erythema: 2
- moderate to severe erythema: 3
- severe erythema (beet redness) to slight eschar formation (injuries in depth): 4

Oedema formation
- no oedema: 0
- very slight oedema (barely perceptible): 1
- slight oedema (edges of area well-defined by definite raising): 2
- moderate oedema (raised approximately 1 millimetre): 3
- severe oedema (raised more than 1 millimetre and extending beyond area of exposure): 4
Any other lesions were noted.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: mean scores after 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no cutaneous reactions

Remarks on result:

other: A brown coloration of the skin, which could have masked a possible very slight erythema (grade 1), was noted at the 1-hour scoring only

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: mean scores after 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no cutaneous reactions

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: mean scores after 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: a very slight or well-defined erythema (grade 1 or 2)

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: mean scores after 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no cutaneous reactions

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: mean scores after 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hors

Remarks on result:

other: a very slight or well-defined erythema (grade 1 or 2)

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: mean scores after 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no cutaneous reactions

Irritant / corrosive response data:

After a 3-minute exposure (one animal):
No cutaneous reactions were observed. A brown coloration of the skin, which could have masked a possible very slight erythema (grade 1), was noted at the 1-hour scoring only.
After a 4-hour exposure (three animals):
No cutaneous reactions were observed in 1/3 animals; a brown coloration of the skin, which could have masked a possible very slight erythema (grade 1), was noted at the 1-hour scoring only.
In both remaining animals, a very slight or well-defined erythema (grade 1 or 2) was noted from day 1 up to day 3.
Mean scores over 24, 48 and 72 hours for each animal were 0.0, 1.0 and 1.0 for erythema and 0.0, 0.0 and 0.0 for oedema.

Interpretation of results:

not irritating

Remarks:

Migrated information according to the Regulation (EC) No. 1272/2008 Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions, the test substance EDDHAS Fe 3K is slightly irritant when applied topically to rabbits. However, the average scores calculated for each of the three animals for three consecutive days (24, 48 and 72 hours) do not exceed cut-off-values defined in the CLP.

Executive summary:

The potential of the test substance Fe3KEDDHSA to induce skin irritation was evaluated in rabbits according to OECD (No. 404, 17th July 1992) and EC (92/69/EEC, B.4, 31st July 1992) guidelines. The test substance was applied for periods of 3 minutes and 4 hours to a single male New Zealand White rabbit. Since the test substance was not irritant on this first animal, it was then applied for 4 hours to two other animals. A single dose of 500 mg of the test substance in its original form was applied to the closely-clipped skin of one flank. The test substance was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing. The mean values of the scores for erythema and oedema were calculated for each animal. After a 3-minute exposure no cutaneous reactions were observed in the first animal. A brown coloration of the skin, which could have masked a possible very slight erythema, was noted at the 1-hour scoring only. After a 4-hour exposure no cutaneous reactions were observed in 1/3 animals; a brown coloration of the skin, which could have masked a possible very slight erythema, was noted at the 1 -hour scoring only. In both remaining animals, a very slight or well-defined erythema was noted from day 1 up to day 3. Mean scores over 24, 48 and 72 hours for each animal were 0.0, 1.0 and 1.0 for erythema and 0.0, 0.0 and 0.0 for oedema. Under the experimental conditions, the test substance Fe3KEDDHSA is slightly irritant when applied topically to rabbits.

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:

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

Adequacy of study:

key study

Study period:

2000-08-17 to 2000-08-25

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EC Directive No. 92/69/EEC, B.5, 31st July 1992

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France
- Age at study initiation: not reported
- Weight at study initiation: mean body weight of 3 animals 3.0 ± 0.1 kg
- Housing: individually in polystyrene cages (48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.
- Diet (e.g. ad libitum): ad libitum. 110 pelleted diet (UAR, 91360 Villemoisson-sur-Orge, France).
- Water (e.g. ad libitum): drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ±3
- Humidity (%): 30 to 70
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg

Duration of treatment / exposure:

single application

Observation period (in vivo):

The eyes were examined approximately 1 hour, 24, 48 and 72 hours after administration of the test substance.

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no


SCORING SYSTEM:

Conjunctival lesions and discharge:
Chemosis (lids and/or nictitating membranes):
- no swelling: 0
- any swelling above normal (includes nictitating membranes): 1
- obvious swelling with partial eversion of lids: 2
- swelling with lids about half-closed: 3
- swelling with lids more than half-closed: 4

Redness (refers to palpebral and bulbar conjunctivae, cornea and iris):
- blood vessels normal: 0
- a number of blood vessels definitely hyperemic (injected): 1
- diffuse, crimson colour, individual vessels not easily discernible: 2
- diffuse, beefy red: 3

Discharge
- absence of discharge: 0
- slight discharge (does not include small amounts normally found in inner canthus): 1
- discharge with moistening of lids and hairs adjacent to lids: 2
- discharge with moistening of lids and hairs on wide area around the eye: 3

Iris lesions
- normal: 0
- markedly deepened rugae, congestion, swelling, moderate circum-comeal hyperemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive): 1
- no reaction to light, haemorrhage, gross destruction (any or all of these): 2

Opacity (degree of intensity: area most dense taken for reading):
- no ulceration or opacity 0
- scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible: 1
- easily discernible translucent area, details of iris slightly obscured: 2
- nacrous areas, no details of iris visible, size of pupil barely discernible: 3
- opaque cornea, iris not discernible through the opacity: 4

Area of opacity:
- one quarter (or less) but not zero: 1
- greater than one quarter but less than a half: 2
- greater than one half but less than three quarters: 3
- greater than three quarters up to whole area: 4

TOOL USED TO ASSESS SCORE: not reported

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

based on 3 animals

Time point:

other: mean value of the scores at each reading (24, 48 and 72h)

Score:

0

Max. score:

4

Reversibility:

other: no opacity observed

Irritation parameter:

iris score

Basis:

mean

Remarks:

based on 3 animals

Time point:

other: mean value of the scores at each reading (24, 48 and 72h)

Score:

0

Max. score:

2

Reversibility:

other: no lesions observed

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

based on 3 animals

Time point:

other: mean value of the scores at each reading (24, 48 and 72h)

Score:

0

Max. score:

3

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: redness and discharge

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean value of the scores at each reading (24, 48 and 72h)

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean value of the scores at each reading (24, 48 and 72h)

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean value of the scores at each reading (24, 48 and 72h)

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritant / corrosive response data:

Slight conjunctival reactions were observed in all animals on day 1: a very slight or slight chemosis (grade 1 or 2), a very slight redness of the conjunctiva (grade 1) and a clear discharge were noted.
A very slight chemosis (grade 1) persisted in 2/3 animals on day 2 and in 1/3 animals on day 3. No other ocular reactions were noted during the study.
Mean scores calculated for each animal over 24, 48 and 72 hours were 0.7, 0.3 and 0.0 for chemosis, 0.0, 0.0 and 0.0 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity.

Other effects:

No other effects were noted

Table 1: Individual ocular examinations and mean values of the scores recorded at each reading (24, 48 and 72 hours) for each animal

Rabbit number	Region of eye	Description of ocular reactions	Scores				Mean irritation score (1)	Interpretation
			lh	24h	48h	72h
			Dl	D2	D3	D4		(-) (+)
753	Conjunctivae	Chemosis	1	1	I	0	0.7	(-)
		Redness	1	0	0	0	0.0	(-)
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0	(-)
	Corneal opacity	Intensity	0	0	0	0	0.0	(-)
		Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		/	U	/	/
769	Conjunctivae	Chemosis	2	1	0	0	0.3	(-)
		Redness	1	0	0	0	0.0	(-)
		Discharge	2	0	0	0	0.0
	Iris		0	0	0	0	0.0	(-)
	Corneal opacity	Intensity	0	0	0	0	0.0	(-)
		Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		/	U	/	/
770	Conjunctivae	Chemosis	1	0	0	0	0.0	(-)
		Redness	1	0	0	0	0.0	(-)
		Discharge	1	0	0	0	0.0
	Iris		0	0	0	0	0.0	(-)
	Corneal opacity	Intensity	0	0	0	0	0.0	(-)
		Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		/	U	/	/
(1) mean of scores on days 2, 3 and 4
h = hour
D = day
(+) = irritant according to E.E.C. criteria
(-) = non-irritant according to E.E.C. criteria
*  = None
U = Fluorescein batch No. C079
/  = Fluorescein not used

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions of this study, the test substance Fe3KEDDHSA is non-irritant when administered by ocular route to rabbits.

Executive summary:

The potential of the test substance Fe3KEDDHSA to induce ocular irritation was evaluated in New Zealand White rabbits according to OECD TG 405. A single dose of 100 mg of the test substance in its original form was introduced into the left conjunctival sac of three animals. The right eye was not treated and served as control. Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. The interpretation of results was carried out according to the classification criteria laid down in the Regulation (EC) No. 1272/2008 (CLP).

Slight conjunctival reactions (very slight or slight chemosis, very slight redness of the conjunctiva and clear discharge) were observed in all animals on day 1 (day of application). A very slight chemosis persisted in 2/3 animals on day 2 (24 -hour reading) and in 1/3 animals on day 3 (48 -hour reading). No lesions were observed on day 3 (72 -hour reading). No other ocular reactions were noted during the study. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.7, 0.3 and 0.0 for chemosis, 0.0, 0.0 and 0.0 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity. The mean scores for 2 out of 3 animals do not exceed trigger values for classification and labelling defined in the CLP. The test substance Fe3KEDDHSA is considered to be not irritating to eyes.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation

The potential of the test substance Fe(3K)EDDHSA (EC No. 462-490-6) to induce skin irritation was evaluated in rabbits according to OECD Guideline 404 (Centre Internationale de Toxicologie, 2000c, Report No. 20503 TAL). The test substance was applied for periods of 3 minutes and 4 hours to a single male New Zealand White rabbit. Since the test substance was not irritant to the first animal, it was then applied for 4 hours to two other animals. A single dose of 500 mg of the test substance in its original form was applied to the closely-clipped skin of one flank. The test substance was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing. The mean values of the scores for erythema and oedema were calculated for each animal. After a 3-minute exposure no cutaneous reactions were observed in the first animal. A brown coloration of the skin, which could have masked a possible very slight erythema, was noted at the 1-hour scoring only. After a 4-hour exposure no cutaneous reactions were observed in 1/3 animals; a brown coloration of the skin, which could have masked a possible very slight erythema, was noted at the 1 -hour scoring only. In both remaining animals, a very slight or well-defined erythema was noted from day 1 up to day 3. Mean scores over 24, 48 and 72 hours for each animal were 0.0, 1.0 and 1.0 for erythema and 0.0, 0.0 and 0.0 for oedema. Under the experimental conditions, the test substance Fe(3K)EDDHSA is slightly irritant when applied topically to rabbits.

Two acute skin irritation/corrosion studies in albino rabbits were performed to determine the irritant or corrosive potency of the test substance Fe(Na)EDDHA (CAS 84539-55-9) on the albino rat skin (CIBA-GEIGY, 1993c, Report No. 931143; Mercier, 1992, Report No. 209306).These tests were based on the OECD Guideline 404 (Acute Dermal Irritation/Corrosion) and were performed on 3 rabbits, checked for normal skin conditions. A gauze patch bearing 0.5 g of the test article was applied to the right flank of each animal and held in place for 4 hours by an adhesive tape. The studies showed that Fe(Na)EDDHA does not cause skin irritation, as the mean values of the recordings 24 to 72 hours after application were scored 0 in the study performed by CIBA-GEIGY (1993) and the scores determined in the study of Mercier (1992) were all below the trigger values for classification as skin irritant.

Eye irritation

The potential of the test substance Fe(3K)EDDHSA (EC No. 462-490-6) to induce ocular irritation was evaluated in New Zealand White rabbits according to OECD Guideline 405 (Centre Internationale de Toxicologie, 2000d, Report No. 20504 TAL). A single dose of 100 mg of the test substance in its original form was introduced into the left conjunctival sac of three animals. The right eye was not treated and served as control. Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. The interpretation of results was carried out according to the classification criteria laid down in the Regulation (EC) No. 1272/2008 (CLP).

Slight conjunctival reactions (very slight or slight chemosis, very slight redness of the conjunctiva and clear discharge) were observed in all animals on day 1 (day of application). A very slight chemosis persisted in 2/3 animals on day 2 (24 -hour reading) and in 1/3 animals on day 3 (48 -hour reading). No lesions were observed on day 3 (72 -hour reading). No other ocular reactions were noted during the study. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.7, 0.3 and 0.0 for chemosis, 0.0, 0.0 and 0.0 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity. The mean scores for 2 out of 3 animals do not exceed trigger values for classification and labelling defined in the CLP. The test substance Fe(3K)EDDHSA is considered to be not irritating to eyes.

Two eye irritation studies were performed according to OECD Guideline 405 (Acute Eye Irritation/Corrosion) with the test item Fe(Na)EDDHA (CAS 84539-55-9) (CIBA-GEIGY, 1993d; Report No. 931144; Mercier, 1992, Report No. 209305). The tests were performed on 3 New Zealand White rabbits. A volume of 0.1 mL of the test item was placed into the conjunctival sac of the left eye of each animal. The lids were then held together for about one second in order to prevent loss of the test article. The animals were checked daily for systemic symptoms and mortality. The studies showed that the test substance does not cause eye irritation as none of the animals achieved a mean score leading to classification according to GHS (>= 1 for cornea and iris; >= 2 for redness and chemosis).

Justification for selection of skin irritation / corrosion endpoint:
Key study conducted with the nearest analogue Fe(3K)EDDHSA (EC 462-490-6)

Justification for selection of eye irritation endpoint:
Key study conducted with the nearest analogue Fe(3K)EDDHSA (EC 462-490-6)

Effects on skin irritation/corrosion: slightly irritating

Justification for classification or non-classification

Based on the results of skin and eye irritation studies available for the structurally related substances Fe(3K)EDDHSA (EC No. 462 -490 -6) and Fe(Na)EDDHA (CAS 84539 -55 -9) the target substance Fe(3Na)EDDHSA is considered to be not irritating to skin and eyes and does need to be classified and labelled as an eye or skin irritant according to Regulation 1272/2008/EC (CLP).