Reaction mass of disodium [2,2'-(imino-κN)dibutanedioato-κ2O1,O4(4-)]calcium(2-), sodium chloride and aspartic acid disodium salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Study period:

from 17-08-1998 till 8-09-1998

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The underlying hypothesis for the read-across is that chelates have the same mode of action based on their ability to chelate, remove or add metal cations to body causing perturbation of body’s micronutrients balance.
The source substance is a chelating agent in a target substance. The only difference between the target and the main source substance is presence of calcium (Ca) cation instead two Na+ cations. As calcium is an essential macro element required by all forms of life, is considered not to influence the toxicological activity.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

2010-11-09 to 2010-11-25

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment. The source substance has the same chelating agent as in a target substance (CaNa2IDHA). The only difference between the target and the source substance is presence of calcium (Ca) cation instead Cu2+ cations. As calcium is an essential macro element required by all forms of life, is considered not to influence the toxicological activity.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: SOP/T/23: “Acute eye irritation/corrosion study

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Slovak National Accreditation Service; Statement of GLP compliance No. G-024

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Experimental Department of National Research Institute of Animal Production, Balice (near Kraków)
- Age at study initiation: Two 8-month-old females (rabbit No 1 and No 2) and one 9-month-old female (rabbit No 3) were used in experiment.
- Weight at study initiation: On day of experiment commencement rabbit No 1 weighed 4.2 kg, rabbit No 2 – 4.6 kg and rabbit No 3 – 3.8 kg.
- Housing: individually in metal cages with dimensions (length x width x height): 60 x 35 x 40 cm.
- Diet (e.g. ad libitum): ad libitum standard granulated "LSK" fodder produced by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL, Motycz
- Water (e.g. ad libitum): ad libitum, tap water.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 – 23
- Humidity (%): 47 – 84
- Air changes (per hr): about 16 times
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
Experiment commencement: 2010-11-09
from 15.11.2010. to 22.11.2010. (rabbit No 1)
from 18.11.2010. to 25.11.2010. (rabbit No 2 and No 3)

Vehicle:

water

Controls:

other: The second eye was used as the control one

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.084 g
- Concentration (if solution): 0.084 g/0.1 mL

Duration of treatment / exposure:

single administration

Observation period (in vivo):

after 1, 24, 48 and 72 hours as well as 7 days after administration of test item, till the moment of retreat of signs in eye.

Number of animals or in vitro replicates:

3

Details on study design:

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

SCORING SYSTEM: Results of detailed clinical observations of animals were recorded according to SOP/T/23 using classification on the basis of OECD Guideline No 405 / EU Method B.5.
Cornea
Opacity: degree of density (readings should be taken from most dense area) *
- No ulceration or opacity: 0
- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
- Easily discernible translucent area, details of iris slightly obscured: 2
- Nacrous area, no details of iris visible, size of pupil barely discernible: 3
- Opaque cornea, iris not discernible through the opacity: 4
Maximum possible: 4
* The area of corneal opacity should be noted

Iris:
- Normal: 0
- Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, iris reactive to light (a sluggish reaction is considered to be an effect): 1
- Hemorrhage, gross destruction, or no reaction to light: 2
Maximum possible: 2

Conjunctivae:
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
- Normal: 0
- Some blood vessels hyperaemic (injected): 1
- Diffuse, crimson colour, individual vessels not easily discernible: 2
- Diffuse beefy red: 3
Maximum possible: 3

Chemosis:
Swelling (refers to lids and/or nictating membranes)
- Normal: 0
- Some swelling above normal: 1
- Obvious swelling, with partial eversion of lids: 2
- Swelling, with lids about half closed: 3
- Swelling, with lids more than half closed: 4
Maximum posible: 4

TOOL USED TO ASSESS SCORE: not reported

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of readings in animal 1, 2 and 3

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: mean of 24, 48, 72 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: mean of 24, 48, 72 hours

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: mean of 24, 48, 72 hours

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #1

Time point:

other: mean of 24, 48, 72 hours

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #2

Time point:

other: mean of 24, 48, 72 hours

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #3

Time point:

other: mean of 24, 48, 72 hours

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean of 24, 48, 72 hours

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean of 24, 48, 72 hours

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean of 24, 48, 72 hours

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

No pathological changes were stated in cornea of eyes. Following administration of test item changes in form of congestion in iris as well as
congestion, swelling and ocular discharge were observed in conjunctiva.

Other effects:

No other effects reported.

Body weights

On day of experiment commencement rabbit No 1 weighed 4.2 kg, rabbit No 2 – 4.6 kg and rabbit No 3 – 3.8 kg. On day of experiment termination rabbit No 1 weighed 4.4 kg, rabbit No 2 – 4.6 kg and rabbit No 3 – 3.7 kg.

Ocular observations

During reading after 1 hour since administration of test item, congestion of iris and normal reaction of pupil to light were stated in animal No 1. Erythema in form of diffuse crimson redness (individual vessels hardly visible) as well as congestion of nictating membrane and circumcorneal injection were stated in conjunctiva in three animals. Furthermore, obvious swelling, with partial eversion of lids, swelling of nictating membrane and ocular discharge on lids and lids’ hair were stated in three animals.

During reading after 24 hours since administration of test item, congestion of iris and normal reaction of pupil to light were still observed in animal No 1. Erythema in form of diffuse crimson redness (individual vessels hardly visible) was still observed in animal No 1 and No 2. Erythema in conjunctiva of animal No 3 intensified: diffuse beefy redness was observed. Furthermore, congestion of nictating membrane and circumcorneal injection were stated in three animals. Obvious swelling, with partial eversion of lids, swelling of nictating membrane and ocular discharge on lids and lids’ hair were still observed in animal No 1 and No 3. Swelling of nictating membrane and slight volumes of ocular discharge were observed in animal No 2.

During reading after 48 hours since administration of test item, no pathological changes were stated in iris of animal No 1. Erythema in form of diffuse crimson redness (individual vessels hardly visible) as well as congestion of nictating membrane and circumcorneal injection were observed in animal No 1 and No 3. Injection of some blood vessels and congestion of nictating membrane were observed in conjunctiva in animal No 2. Furthermore, slight swelling of conjunctiva was stated in animal No 3. Swelling of nictating membrane was stated in three animals. Slight volumes of ocular discharge was stated in animal No 1, ocular discharge on lids and lids’ hair was stated in animal No 3. No ocular discharge was observed in animal No 2.

During reading after 72 hours since administration of test item, injection of blood vessels and congestion of nictating membrane were observed in conjunctiva in animal No 1 and No 2. Erythema in form of diffuse crimson redness (individual vessels hardly visible) as well as congestion of nictating membrane and circumcorneal injection were still observed in conjunctiva in animal No 3. Furthermore, slight swelling, swelling of nictating membrane and slight volumes of ocular discharge were observed in animal No 3. No swelling and ocular discharge were observed in animal No 1 and No 2.

During reading after 7 days since administration of test item, no pathological changes were stated in eye of three animals.

Table 1. Point evaluation of acute eye irritation / corrosion

Rabbit No	Part of eye		Reading after					Average of readings after 24, 48, 72 hours
			1 hour	24 hours	48 hours	72 hours	7 days
1	Cornea		0	0	0	0	0	0.0
	Iris		1	1	0	0	0	0.3
	Conjunctiva	erythema	2	2	2	1	0	1.7
		swelling	2	2	1	0	0	1.0
2	Cornea		0	0	0	0	0	0.0
	Iris		0	0	0	0	0	0.0
	Conjunctiva	erythema	2	2	1	1	0	1.3
		swelling	2	1	1	0	0	0.7
3	Cornea		0	0	0	0	0	0.0
	Iris		0	0	0	0	0	0.0
	Conjunctiva	erythema	2	3	2	2	0	2.3
		swelling	2	2	1	1	0	1.3

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the ground of performed study and according to Annex to Decree of Ministry of Health of September 4th, 2007 (Acts Daily No 174, Position 1222) one may state that Cu (II) IDHA does not irritate eye of rabbit.
According to Regulation (EC) No. 1272/2008, Cu (II) IDHA does not need to be classified and labelled for eye irritation effects as all scores except one score of 2.3 for conjunctival erythema were under the cut-off values triggering C&L.

Executive summary:

The acute eye irritation/corrosion study was performed in order to obtain information on health risk of Cu (II) IDHA in eyes of three (females) New Zealand White rabbits (Kropidło, 2010; Project No. ODR – 13/10). The study was commenced with the sighting study on one animal. The powdered test item was administered as a single dose in amount of 0.084 g (it was volume of 0.1 mL) to conjunctival sack of one eye of animal (rabbit No 1). The second eye served as the control. The animal was observed for 7 days till the moment of retreat of signs in eye. After evaluation of treated eye, in order to confirm irritation or its lack, the test item was given to eye of another two animals (rabbit No 2 and No 3). Manner of proceeding was the same as in case of rabbit No 1. The animals were observed for period for 7 days, till the moment of retreat of signs in eye. General clinical observations for morbidity and mortality were performed in animals daily during the entire experiment. Detailed clinical observations for changes in cornea, iris and conjunctiva were performed after 1, 24, 48 and 72 hours as well as after 7 days since administration of test item. Body weight of animals was determined directly before administration on day of test item administration (day 0) and on the last day of experiment. After period of observation the animals were humanely sacrificed.

No mortalities or clinical signs were noted in treated animals. Following administration of test item no pathological changes were stated in cornea of eyes. Changes in form of congestion in iris in one animal as well as congestion, swelling and ocular discharge were observed in conjunctivae of three animals. In details, regarding erythema in conjunctivae, transiently effects were noted: diffuse crimson redness (in three animals) , diffuse beefy redness (in one animal) and injection of blood vessels (in two animals). Further transient effects were congestion of nictating membrane and circumcorneal injection in three animals. Regarding swelling in conjunctivae, transient effects were noted in three animals: first obvious with partial eversion of lids, then slight swelling as well as swelling of nictating membrane. Among other transient effects, discharge on lids and lids’ hair was noted in three animals. All effects were fully reversible within 7 days.

On the ground of performed study and according to Annex to Decree of Ministry of Health of September 4th, 2007 (Acts Daily No 174, Position 1222) one may state that Cu (II) IDHA does not irritate eye of rabbit.

Cross-reference 2

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

23 of May till 27 of June 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment. The source substance has the same chelating agent as in a target substance (CaNa2IDHA). The only difference between the target and the source substance is presence of calcium (Ca) cation instead Fe3+ cations. As calcium is an essential macro element required by all forms of life, is considered not to influence the toxicological activity.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: conventional Reproductive Farm of Rabbits, Balice
- Age at study initiation: 6-month-old
- Weight at study initiation: 3,60 kg
- Housing:
- Diet (e.g. ad libitum): standard granulated fodder "LSK" produced by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL, Motycz
- Water (e.g. ad libitum): tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 – 22
- Humidity (%): 52 - 70% (few times above 70%);
- Photoperiod (hrs dark / hrs light): artificial fluorescent lighting: 12 hours light / 12 hours darkness

The animals were kept individually in metal cages with dimensions: 60 x 35 x 40 cm. Each cage was equipped with label containing information on study code, start date of the experiment, sex and animal numbers.

IN-LIFE DATES: The test material was administered to rabbits’ eyes on: 31.05.2007 (rabbit No 1) and 04.06.2007 (rabbits No 2 and 3). The experiment was terminated on 07.06.2007 (rabbit No 1 and 2) and 14.06.2007 (rabbit No 3).

Vehicle:

unchanged (no vehicle)

Controls:

other: The second eye was used as the control one.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g of unchanged test substance as a single
- Concentration (if solution): not applied

Duration of treatment / exposure:

single administration

Observation period (in vivo):

After 1, 24, 48,72 hours and 7 days since administration of the test material condition
of cornea, iris and conjunctiva was evaluated.

Number of animals or in vitro replicates:

3

Details on study design:

The test material in form of powder was administered in amount of 0,1 g as a single dose to conjunctival sack of one eye to one rabbit (rabbit No 1), by gentle distraction of lower eyelid. Then both eyelids were kept together for a moment to protect loosing of the test material. The second eye was used as the control one. Following evaluation of the treated eye, in order to confirm the obtained results, the test material was administered in the same way to rabbits No 2 and 3.

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

SCORING SYSTEM: classification on the basis of OECD Guideline No 405 / EU Method B.5
Cornea
Opacity: degree of density (readings should be taken from most dense area) *
- No ulceration or opacity: 0
- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
- Easily discernible translucent area, details of iris slightly obscured: 2
- Nacrous area, no details of iris visible, size of pupil barely discernible: 3
- Opaque cornea, iris not discernible through the opacity: 4
Maximum possible: 4
* The area of corneal opacity should be noted
Iris:
- Normal: 0
- Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, iris reactive to light (a sluggish reaction is considered to be an effect): 1
- Hemorrhage, gross destruction, or no reaction to light: 2
Maximum possible: 2
Conjunctivae:
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
- Normal: 0
- Some blood vessels hyperaemic (injected): 1
- Diffuse, crimson colour, individual vessels not easily discernible: 2
- Diffuse beefy red: 3
Maximum possible: 3

Chemosis:
Swelling (refers to lids and/or nictating membranes)
- Normal: 0
- Some swelling above normal: 1
- Obvious swelling, with partial eversion of lids: 2
- Swelling, with lids about half closed: 3
- Swelling, with lids more than half closed: 4
Maximum posible: 4

TOOL USED TO ASSESS SCORE: not recorded

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #1

Time point:

other: average after 24 - 72 h

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

oedema

Basis:

animal #1

Time point:

other: average after 24- 72h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #2

Time point:

other: average after 24-72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 48h

Irritation parameter:

conjunctivae score

Remarks:

oedema

Basis:

animal #2

Time point:

other: average after 24-72 h

Score:

0.3

Max. score:

2

Reversibility:

fully reversible within: 48h

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #3

Time point:

other: average after 24-72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

oedema

Basis:

animal #3

Time point:

other: average after 24-72h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Irritant / corrosive response data:

During observations only changes in eye conjunctiva were stated. During evaluation after 1 hour since the test material administration, all rabbits showed erythema of eye conjunctiva – in form of diffuse crimson redness, congestion of third eyelid and circumcorneal injections. Rabbits No 1 and 3 showed slight
oedema while rabbit No 2 distinct oedema with partial eversion of eyelids. Additionally all rabbits showed oedema of third eyelid and excretion – on eyelids and
eyelids hair in rabbits no 1 and 2 and slight amount of excretion in rabbit No 3.
During evaluation after 24 hours since the administration, all rabbits showed congestion of third eyelid, circumcorneal injection and erythema of eye conjunctiva – in form of diffuse crimson redness in rabbits No 1 and 2 and in form of blood vessels hyperaemia (injection) in rabbit No 3. In all rabbits slight oedema of third eyelid was stated. Additionally all rabbits showed excretion – on eyelids and eyelids hair in rabbit No 1 and slight amount of excretion in rabbits No 2 and 3.
During observation after 48 hours since administration, all rabbits showed erythema – in form of diffuse crimson redness in rabbit No 1 and in form of blood vessels congestion (injection) in rabbits No 2 and 3. Additionally rabbit No 1 showed circumcorneal injection, oedema of third eyelid and excretion on eyelids and eyelids hair.
During evaluation after 72 hours since administration rabbits No 1 and 3 showed erythema in form of blood vessels congestion (injection) and congestion of third eyelid. Additionally rabbit No 1 showed circumcorneal injection, oedema of third eyelid and excretion on eyelids and eyelids hair. Rabbit No 2 showed no pathological changes.
During evaluation after 7 days since administration no pathological changes were stated in eyes of rabbits No 1 and 3.

Other effects:

At the start of the experiment the rabbits weighed: 3,73 kg (rabbit No 1), 3,56 kg (rabbit No 2) and 3,52 kg (rabbit No 3). At the end of the test body weight of rabbits amounted appropriately: 3,84 kg; 3,60 kg and 3,54 kg.

Chelate with name: Fe (III) IDHA - Point evaluation of acute eye irritation/corrosion study for rabbits

Rabbit No	Part of eye		Evaluation after					Average of evaluation after 24, 48 and 72 h
			1h	24h	48h	72h	7 days
1	cornea		0	0	0	0	0	0
	Iris		0	0	0	0	0	0
	conjunctiva	erythema	2	2	2	1	0
		oedema	1	1	1	1	0
2	cornea		0	0	0	0	-	0
	Iris		0	0	0	0	-	0
	conjunctiva	erythema	2	2	1	0	-
		oedema	2	1	0	0	-
3	cornea		0	0	0	0	0	0
	Iris		0	0	0	0	0	0
	conjunctiva	erythema	2	1	1	1	0
		oedema	1	1	0	0	0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Fe (III) IDHA does not irritate eyes of the rabbits.

Executive summary:

Acute eye irritation/corrosion study of Chelate with name: Fe (III) IDHA for rabbits was performed according to OECD Guideline for Testing of Chemicals No 405/Method B.5. as well as Principles of Good Laboratory Practice (GLP - OECD,1997).

The study was performed with three white rabbits of New Zealand strain. The test material in form of powder was administered in amount of 0.1 g to conjunctival sack of one eye to each rabbit. After 1, 24, 48, 72 hours and 7 days since administration of

the test material, condition of cornea, iris and conjunctiva was evaluated. During evaluation after 1, 24, 48 and 72 the test rabbits showed changes in form of erythema and oedema of eye conjunctiva.

Classification of the test material was performed according to Regulation 1272/2008. Taking into account the obtained results, one may state that Chelate with name: Fe (III) IDHA does not irritate eyes of rabbits.

Cross-reference 3

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

2013-11-08 to 2013-11-16

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment. The source substance has the same chelating agent as in a target substance (CaNa2IDHA). The only difference between the target and the source substance is presence of calcium (Ca) cation instead Mn2+ cations. As calcium is an essential macro element required by all forms of life, is considered not to influence the toxicological activity.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: SOP/T/23: “Acute eye irritation/corrosion study”

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bureau for chemical substances, Registration No. 8/2012/DPL

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: the Experimental Department of the National Research Institute of Animal Production, Balice near Kraków.
- Age at study initiation: 8.5-month-old – rabbit No. 1 and 6-mounths-old – rabbit No. 2 and No. 3.
- Weight at study initiation: At the start of the experiment rabbit No. 1 weighed: 4.0 kg, rabbit No. 2 weighted 3.9 kg and rabbit No. 3 weighted 4.1 kg.
- Housing: individually in metal cages with dimensions (length x width x height): 60 x 35 x 40 cm.
- Diet (e.g. ad libitum): ad libitum; standard granulated "LSK" fodder produced by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL, Motycz
- Water (e.g. ad libitum): ad libitum; tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22
- Humidity (%): 45 – 60
- Air changes (per hr): about 16 times/h
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
Start of the study: 04.11.2013
Start of the experimental phase: 08.11.2013
End of the experimental phase: 16.11.2013

Vehicle:

water

Controls:

other: The second eye was used as the control one

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.065 g
- Concentration (if solution):0.065 g/0.1 mL

Duration of treatment / exposure:

single administration

Observation period (in vivo):

1, 24, 48 and 72 hours after administration of the test item

Number of animals or in vitro replicates:

3

Details on study design:

Sixty minutes prior to the test item application, an analgesic, i.e. bunondol at a dose of 0.01 mg/kg b.w. was administered to each animal. Five minutes prior to the test item application, one or two drops of a topical anesthetic alcaine was applied to both eyes. During reading after 1 hour after administration of the test item, no corneal opacity and no lesions of iris in the treated eye were stated and redness and swelling were not more than 2, so the administration of anesthetics was discontinued.

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

SCORING SYSTEM: classification on the basis of OECD Guideline No 405 / EU Method B.5
Cornea
Opacity: degree of density (readings should be taken from most dense area) *
- No ulceration or opacity: 0
- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
- Easily discernible translucent area, details of iris slightly obscured: 2
- Nacrous area, no details of iris visible, size of pupil barely discernible: 3
- Opaque cornea, iris not discernible through the opacity: 4
Maximum possible: 4
* The area of corneal opacity should be noted

Iris:
- Normal: 0
- Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, iris reactive to light (a sluggish reaction is considered to be an effect): 1
- Hemorrhage, gross destruction, or no reaction to light: 2
Maximum possible: 2

Conjunctivae:
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
- Normal: 0
- Some blood vessels hyperaemic (injected): 1
- Diffuse, crimson colour, individual vessels not easily discernible: 2
- Diffuse beefy red: 3
Maximum possible: 3

Chemosis:
Swelling (refers to lids and/or nictating membranes)
- Normal: 0
- Some swelling above normal: 1
- Obvious swelling, with partial eversion of lids: 2
- Swelling, with lids about half closed: 3
- Swelling, with lids more than half closed: 4
Maximum posible: 4

TOOL USED TO ASSESS SCORE: not reported

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of readings in animal 1, 2 and 3

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Irritation parameter:

iris score

Basis:

mean

Remarks:

of readings in animal 1, 2 and 3

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #1

Time point:

other: mean of 24, 48 and 72 hours

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #2

Time point:

other: mean of 24, 48 and 72 hours

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

animal #3

Time point:

other: mean of 24, 48 and 72 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean of 24, 48 and 72 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Irritant / corrosive response data:

After the application of the test item, pathological changes in the conjunctivas of the animals’ eyes were stated. No pathological changes in the cornea and iris were stated.

Other effects:

No other effects reported.
At the start of the experiment rabbit No. 1 weighed: 4.0 kg, rabbit No. 2 weighted 3.9 kg and rabbit No. 3 weighted 4.1 kg. On the day of experimental end the animals weighed respectively: 4.0 kg, 3.8 kg and 4.1 kg.

Body weight

At the start of the experiment rabbit No. 1 weighed: 4.0 kg, rabbit No. 2 weighted 3.9 kg and rabbit No. 3 weighted 4.1 kg. On the day of experimental end the animals weighed respectively: 4.0 kg, 3.8 kg and 4.1 kg.

Ocular findings

After the application of the test item, pathological changes in the conjunctivas of the animals’ eyes were stated. No pathological changes in the cornea and iris were stated. During the reading which took place 1 hour after the application of the test item, congestion of some blood vessels was stated in rabbit No. 1 and 3, whereas diffuse crimson colour was stated in rabbit No. 2. Congestion of the nictitating membrane, circumcorneal injection, slight swelling of the conjunctiva, and swelling of the nictitating membrane were stated in three rabbits. Furthermore, a small volume of discharge in rabbit No. 1 and 3, and discharge on the lids and lids’ hair in rabbit No. 2 were observed.

During the reading which took place 24 hours after the application of the test item, congestion of some blood vessels was stated in rabbits No. 1 and 3, whereas diffuse, crimson colour was stated in rabbit No. 2. Congestion of the nictitating membrane was stated in three rabbits. Additionally, circumcorneal injection was stated in rabbit No. 1. Slight swelling of the conjunctiva and swelling of the nictitating membrane were still observed in rabbit No. 1. No swelling was stated in rabbits No. 2 and 3. No discharge from the conjunctivae sack was stated in three rabbits. During the reading which took place 48 hours after the application of the test item, congestion of some blood vessels and congestion of the nictitating membrane were stated in rabbit No. 1 and 2. Additionally, circumcorneal injection in rabbit No. 1 was still observed. No pathological changes were stated in rabbit No. 3. During the reading which took place 72 hours after the application of the test item, no pathological changes in the conjunctivas of the rabbits were stated. The detailed results of the clinical observations are presented in Table 1.

Table 1. Detailed results of clinical observations of eye

Animal	Reading after	CHANGES IN EYE
No		CORNEA	IRIS	CONJUNCTIVA
				erythema	swelling	other
1	1h	no changes	no changes	injection of some blood vessels; congestion of nictating membrane, circumcorneal injection	slight swelling; swelling of nictating membrane	slight volume of discharge
	24h	no changes	no changes	injection of some blood vessels; congestion of nictating membrane; circumcorneal injection	slight swelling; swelling of nictating membrane	no changes
	48h	no changes	no changes	injection of some blood vessels; congestion of nictating membrane; circumcorneal injection	no changes	no changes
	72h	no changes	no changes	no changes	no changes	no changes
2	1h	no changes	no changes	diffuse crimson colour; congestion of nictating membrane; circumcorneal injection	slight swelling; swelling of nictating membrane	discharge on lids and lids' hair
	24h	no changes	no changes	diffuse crimson colour; congestion of nictating membrane,	no changes	no changes
	48h	no changes	no changes	injection of some blood vessels; congestion of nictating membrane	no changes	no changes
	72h	no changes	no changes	no changes	no changes	no changes
3	1h	no changes	no changes	injection of some blood vessels, congestion of nictating membrane, circumcorneal injection	slight swelling; swelling of nictating membrane	slight volume of discharge
	24h	no changes	no changes	injection of some blood vessels, congestion of nictating membrane	no changes	no changes
	48h	no changes	no changes	no changes	no changes	no changes
	72h	no changes	no changes	no changes	no changes	no changes

The point evaluation of acute eye irritation/corrosion based on the scoring system given in the OECD Guideline No 405 / EU Method B.5. is presented in Table 2.

Table 2. Point evaluation of acute eye irritation / corrosion

Rabbit No	Part of eye		Reading after				Average of readings after 24, 48, 72 hours
			1 hour	24 hours	48 hours	72 hours
1	Cornea		0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Conjunctiva	erythema	1	1	1	0	0.7
		swelling	1	1	0	0	0.3
2	Cornea		0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Conjunctiva	erythema	2	2	1	0	1.0
		swelling	1	0	0	0	0.0
3	Cornea		0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Conjunctiva	erythema	1	1	0	0	0.3
		swelling	1	0	0	0	0.0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the ground of the study the test item Mn (II) IDHA:
- it does not irritate the rabbit eye – according to the Annex to the Decree of the Minister of Health of August 10, 2012 on classification of chemical substances and mixtures (Journal of Laws, item 1018),
- it is beyond the categorization – according to the Regulation (EC) of the European Parliament and of the Council No 1272/2008 of 16 December, 2008 on classification, labeling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC and amending Regulation (EC) No 1907/2006.

Executive summary:

The acute eye irritation/corrosion study was performed in order to obtain information on health risk of Mn (II) IDHA in eyes of three (females) New Zealand White rabbits (Somat, 2013; Project No. ODR – 13/11). The study was commenced with the sighting study on one animal. The powdered test item was administered as a single dose in amount of 0.065 g (it was volume of 0.1 mL) to conjunctival sack of one eye of animal (rabbit No 1). The second eye served as the control. The animal was observed for 72 hours. After evaluation of the treated eye, the test item was given to eyes of another two animals (rabbit No 2 and No 3). Manner of proceeding was the same as in case of rabbit No 1. The animals were observed for period for 72 hours. General clinical observations for morbidity and mortality were performed in animals daily during the entire experiment. Detailed clinical observations for changes in cornea, iris and conjunctiva were performed after 1, 24, 48 and 72 hours since administration of test item. Body weight of animals was determined directly before administration on day of test item administration (day 0) and on the last day of experiment. After period of observation the animals were humanely sacrificed.

No mortalities or clinical signs were noted in treated animals. Following administration of test item no pathological changes were stated in cornea and iris of treated eyes. The conjunctival changes were present in three animals. They included erythema and swelling: injection of some blood vessels; congestion of nictating membrane; circumcorneal injection and crimson colour, slight swelling of conjunctivae, swelling of nictating membrane, slight volume of discharge and slight discharge on lids and lid’s hair. The changes were transient:

Administered volume of test item		0.1 mL
Number of rabbit		1	2	3
Sex		female	female	female
Mortality		0/1	0/1	0/1
General clinical signs		none	none	none
PART OF EYE		Changes in eye
CORNEA		none	none	none
IRIS		none	none	none
CONJUNCTIVA	erythema	Transient: injection of some blood vessels; congestion of nictating membrane; circumcorneal injection	Transient: diffuse, crimson colour; injection of some blood vessels; congestion of nictating membrane; circumcorneal injection	Transient: injection of some blood vessels; congestion of nictating membrane; circumcorneal injection
	swelling	Transient: slight swelling; swelling of nictating mebrane	Transient: slight swelling; swelling of nictating mebrane	Transient: slight swelling; swelling of nictating mebrane
	other	Slight volume of discharge	Discharge on lids and lids' hair	Slight volume of discharge

On the ground of the study the test item Mn (II) IDHA:

- it does not irritate the rabbit eye – according to the Annex to the Decree of the Minister of Health of August 10, 2012 on classification of chemical substances and mixtures (Journal of Laws, item 1018),

- it is beyond the categorization – according to the Regulation (EC) of the European Parliament and of the Council No 1272/2008 of 16 December, 2008 on classification, labeling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC and amending Regulation (EC) No 1907/2006.

Cross-reference 4

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

18-08-2002 till 23-09-2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment. The source substance has the same chelating agent as in a target substance (CaNa2IDHA). The only difference between the target and the source substance is presence of calcium (Ca) cation instead Zn2+ cations. As calcium is an essential macro element required by all forms of life, is considered not to influence the toxicological activity.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Principles of method if other than guideline:

: W trakcie badania wilgotność względna powietrza kilkakrotnie przekroczyła wartość 70 %. Nie miało to wpływu na przebieg i wyniki badania. Nie stwierdzono innych odchyleń od wytycznej / metody

GLP compliance:

yes (incl. QA statement)

Remarks:

G-024

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

-----Zwierzęta.-----

W doświadczeniu użyto trzech królików białych rasy nowozelandzkiej
pochodzących z Instytutu Zootechniki w Balicach.
Króliki przeszły minimum 5-dniowy okres kwarantanny, w czasie którego były pod
stałą obserwacją [SPR/T/6].
W dniu przyjęcia zwierząt do kwarantanny wykonano ogólne badanie lekarsko-
weterynaryjne, a przed wprowadzeniem do doświadczenia u zwierząt wykonano
szczegółowe badanie lekarsko-weterynaryjne [SPR/T/46]. Do doświadczenia
wprowadzono zwierzęta nie wykazujące żadnych objawów klinicznych.
Wszystkie zwierzęta były indywidualnie oznaczone [SPR/T/7].
W doświadczeniu użyto trzech królików: jednego samca w wieku 5 miesięcy
(królik nr 1) i dwóch samic w wieku w wieku 6,5 miesięca (królik nr 2 i nr 3).

-----Warunki przetrzymywania zwierząt.-----

W okresie kwarantanny i doświadczenia zwierzęta przebywały w klimatyzowanym
pomieszczeniu o następujących parametrach:
- temperatura powietrza 20 – 22 °C
- wilgotność względna powietrza 50 – 90 %
- oświetlenie sztuczne, jarzeniowe; cykl oświetlenia: 12 godzin jasno - 12 godzin
ciemno [SPR/T/9].
Zwierzęta przetrzymywano pojedynczo w metalowych klatkach, o wymiarach
(długość x szerokość x wysokość): 60×35×40 cm. Każda klatka wyposażona była w
wywieszkę zawierającą: kod badania, datę założenia i likwidacji doświadczenia, płeć i numer zwierzęcia [SPR/T/23].


-----Pasza i woda.-----

Zwierzętom podawano bez ograniczeń granulowaną standardową paszę
laboratoryjną „LSK”, produkowaną przez Wytwórnię Koncentratów i Mieszanek
Paszowych AGROPOL z Motycza oraz wodę wodociągową [SPR/T/15].

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

Badany materiał, po roztarciu na proszek, w ilości 0,100 g (co stanowiło objętość
0,1 ml) podano jednorazowo jednemu królikowi do worka spojówkowego jednego oka,
odciągając delikatnie dolną powiekę. Następnie przez chwilę przytrzymywano obie
powieki, aby zapobiec wydostawaniu się badanego materiału na zewnątrz. Drugie oko
służyło jako kontrola.
Po ocenie narażanego oka, w celu potwierdzenia wyników badania na jednym
zwierzęciu, badany materiał został podany w taki sam sposób jak u królika nr 1, do oka
królika nr 2 i nr 3 [SPR/T/23].

Duration of treatment / exposure:

Jednokrotne podanie.

Observation period (in vivo):

Obserwacji rogówki, tęczówki i spojówki oka dokonano po 1, 24, 48, 72
godzinach oraz po 7 dniach od momentu podania badanego materiału [SPR/T/23].
Oceny dokonano w oparciu o skalę punktową według Wytycznej OECD nr 405 /
Metody B.5. (Załącznik 7).

Number of animals or in vitro replicates:

3 sztuki

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1 h

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24 h

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 48 h

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 72 h

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 7 dni

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 h

Score:

1

Max. score:

1

Reversibility:

not specified

Irritation parameter:

iris score

Basis:

mean

Time point:

other: h

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 7 dni

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

rumień

Basis:

mean

Time point:

other: 1 h

Score:

2

Max. score:

2

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

rumień

Basis:

mean

Time point:

other: 1 h

Score:

2

Max. score:

2

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

rumień

Basis:

mean

Time point:

other: 72 h

Score:

1

Max. score:

1

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

rumień

Basis:

mean

Time point:

other: 7 dni

Score:

0

Max. score:

0

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

obrzęk

Basis:

mean

Time point:

other: 1 h

Score:

2

Max. score:

2

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

obrzęk

Basis:

mean

Time point:

other: 72 dni

Score:

1

Max. score:

1

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Remarks:

obrzęk

Basis:

mean

Time point:

other: 7 dni

Score:

0

Max. score:

0

Reversibility:

not specified

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: POLAND : Rozporządzenia Ministra Zdrowia z dnia 2 września 2003 r. w sprawie kryteriów i sposobu klasyfikacji substancji i preparatów chemicznych (Dz.U. Nr 171 poz. 1666)

Executive summary:

Na podstawie przeprowadzonego badania i według Załącznika do Rozporządzenia Ministra Zdrowia z dnia 2 września 2003 r. w sprawie kryteriów i sposobu klasyfikacji substancji i preparatów chemicznych (Dz.U. Nr 171 poz. 1666) można stwierdzić, że badany materiał Zn (II) IDHA nie drażni oczu królików.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

tetrasodium;2-(1,2-dicarboxylatoethylamino)butanedioate

Cas Number:

144538-83-0

Molecular formula:

C8H7NNa4O

IUPAC Name:

tetrasodium;2-(1,2-dicarboxylatoethylamino)butanedioate

Test material form:

solid

Test animals / tissue source

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

Speeiesi strain/ stock/breeder: rabbit/Himalayan/CHR. FRED LEUSCHNER & CO. D-24601 Lohndorf/Post Wankendorf
Number of animals examined: 3 male rabbits
lnitial age: approx. 4 months
lnitial body weight : 2.0 - 2.4 kg
ldentification: by tattooed number assigned by the Lohndorf breeding station and continuous number 1 - 3
Duration of study: at least 20 adaptation days, 1 test day and a follow-up period of 72 hours

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

100 mg

Observation period (in vivo):

72 h

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

Conduct
A dose of 100 mg lminodisuccinic acid, sodium salt (IDS, Na-Salz) was administered into the conjunctival sac of the right eye of rabbits alter gently pulling the lower lid away from the eyeball. The lid was then gently held together for about one second in order to prevent loss of test material.
The left eye, which remained untreated, served as a control.
Alter the administration the animals were kept separately in special restrainers which allowed free movement of the head but prevented a complete body turn, wiping of the eyes by the paws and excluded irritation of the eyes by excrements and urine.

Examination of the eyes: the eyes were examined ophthalmoscopically with a slit lamp prior to the administration and also 1, 24, 48, 72 hours alter the administration. The eye reactions were observed and registered.
24 hours alter administration the eyes were treated additionally with fluorescein1 and examined.
The effects observed were graded according to the scheme given in Appendix 1 to this report.
Based on most recent guidelines the eye reactions are monitored until the changes observed have completely subsided, however for not more than 21 days alter application. Animals that do not reveal any lesions anymore for 24 hours following the first 72 hours of observation are sacrificed.

General criteria: body weight of all animals was measured at the beginning of the study.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 1,2,3

Remarks:

for all animals was the same result

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

animal: 1,2,3,

Remarks:

for all animals was the same result

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 1,2,3

Remarks:

for all animals was the same result

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

Under the present test conditions a single application of 100 mg lminodisuccinic acid, sodium salt (IDS, Na-Salz} per animal into the conjunctival sac of the right eye of three rabbits did not cause any changes.

The cornea, iris and conjunctivae were not affected by instillation of the test com­ pound.

There were no systemic intolerance reactions.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The tested substance lminodisuccinic acid, sodium salt (IDS, Na-Salz) is not irritant to eyes.

Executive summary:

Test system: acute eye irritation study by instillation into the coniunctival sac of rabbits according to EC guideline B.5. and OECD guideline 405.

Test substance: lminodisuccinicacid,sodiumsalt (IDS,Na-Salz).

Under the present test conditions a single application of 100mg lminodisuccinic acid, sodium salt (IDS,Na-Salz) per animal into the conjunctival sac of the right eye of three rabbits did not cause anychanges. The cornea,iris and conjunctivae were not affected by instillation of the test compound.

There were no systemic intolerance reactions.

According to the EC-Commission directive67/548/EEC and its subsequent amend­ ments on the approximation of the laws, regulations and administrative provision relating to the classification, packaging and labelling of dangerous substances and the results obtained under the present test conditions lminodisuccinic acid, sodium salt {IDS, Na-Salz) was non-irritating to eyes, hence, no labelling is required.

.