Reaction mass of disodium [2,2'-(imino-kappaN)dibutanedioato-kappa2O1,O4(4-)]manganese(2-) and sodium sulphate

Endpoint summary

Description of key information

- Skin irritation: OECD 404, 3 New Zealand White rabbits: barely perceptible erythema in 2 animals after 1 hours and in one animal after 24 hours. Conclusion: not irritating to the skin.
- Eye irritation: OECD 405, 3 New Zealand White rabbits: no corneal opacity and iritis findings; conjunctival congestion, swelling and ocular discharge. No positive responders. Conclusion: not irritating 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:

experimental study

Adequacy of study:

key study

Study period:

2013-11-06 to 2013-11-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well-documented GLP-Guideline Study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: SOP/T/22: “Acute skin irritation/corrosion study”

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bureau for chemical substances (Registration No. B/2012/DPL

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Experimental Department of National Research Institute of Animal Production, Balice near Kraków
- Age at study initiation: 6-month-old – rabbit No. 1 and 6.5-month-old – rabbit No. 2, and on one 9-month-old female – rabbit No. 3
- Housing: individually in metal cages (60 x 35 x 40 cm (length x width x height)). Each cage was equipped with a label containing the number of the study protocol, the dates of the experiment commencement and termination, and sex and number of the animal.
- 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): 20 - 22;
- Humidity (%): 45 – 65
- Air changes (per hr): about 16 times/h
- Photoperiod (hrs dark / hrs light): 12 / 12

Start of the study: 04.11.2013
Start of the experimental phase: 06.11.2013
End of the experimental phase: 11.11.2013

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5 g (powdered test item)

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48, and 72 hours after the end of the exposure

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 6 cm²
- Type of wrap if used: The gauze patches were covered with PCV foil and fixed using a non-irritating sticking plaster

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test item was removed using water
- Time after start of exposure: 4 hours

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: mean at 24, 48 and 72 hours

Score:

0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: mean at 24, 48 and 72 hours

Score:

0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: mean at 24, 48 and 72 hours

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: mean at 24, 48 and 72 hours

Score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: mean at 24, 48 and 72 hours

Score:

0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: mean at 24, 48 and 72 hours

Score:

0

Irritant / corrosive response data:

During the reading which took place 1 hour after the end of the exposure, no pathological changes on the treated skin of rabbit No. 1 were stated. Very slight (barely perceptible) erythema was stated in rabbit No. 2 and 3.
During the reading which took place 24 hours after the end of the exposure, no pathological changes were stated in rabbits No. 1 and 2. Very slight (barely perceptible) erythema was still observed in rabbit No. 3.
During the reading which took place 48 and 72 hours after the end of the exposure, no pathological changes on the treated skin of the rabbits were stated.

Table 1: Summary of results:

Administered volume of test item	0.5 mL
Number of rabbit	1	2	3
Sex	male	male	female
Mortality of animals	0/1	0/1	0/1
General clinical signs	none	none	none
Skin erythema in the treatment area	none	very slight (barely perceptible) after 1 hour	very slight (barely perceptible) after 1 hour and 24 hours
Skin oedema in the treatment area	none	none	none
Other	none	none	none

Body weight of the animals

At the start of the experiment, animal No. 1 weighed 3.3 kg, animal No. 2 weighed 3.6 kg and animal No. 3 weighed 4.3 kg. On the last day of the experiment, animal No. 1 weighed 3.4 kg, animal No. 2 weighed 3.7 kg and animal No. 3 weighed 4.3 kg.

Interpretation of the study results

On the grounds of the study, the test item, i.e. Mn (II) IDHA can be classified to the following categories:

- does not irritate the rabbit skin – 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),

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

The detailed results of the study based on the scoring system presented in the OECD Guideline No. 404 / EU Method B.4. are given in Table 2.

Table 2 - Point evaluation of acute skin irritation/corrosion on rabbits
Acute skin irritation/corrosion on rabbits
Cu (II) IDHA
Animal No	Observation type	Evaluation after				Average after24, 48and72hours
		1hour	24hours	48hours	72hours
1	erythema	0	0	0	0	0
	oedema	0	0	0	0	0
2	erythema	1	0	0	0	0
	oedema	0	0	0	0	0
3	erythema	1	1	0	0	0.3
	oedema	0	0	0	0	0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The observations of the treated skin revealed some changes in the form of barely perceptible erythema. The test item Mn (II) IDHA does not irritate skin of rabbits.

Executive summary:

A study was undertaken to investigate the skin irritation potential of Mn (II) IDHA in New Zealand White rabbits (OECD 404; Sornat, 2013, Report No. DDR-15/13). The study commenced with a sighting study on one animal. The powdered test item was applied in an amount of 0.5 g once to the shaved skin of one animal (rabbit No. 1) and covered with an appropriate band. The exposure lasted 4 hours. After evaluation of treated skin, in order to confirm irritation or its absence, the test item was applied to the skin of the next two animals (rabbit No. 2 and No. 3) for 4 hours in order to confirm its irritant properties. The procedure was the same as in case of rabbit No. 1. General clinical observations of the animals for morbidity and mortality were performed daily during the entire experiment. Detailed clinical observations of the treated skin were performed 1, 24, 48, and 72 hours after the end of the exposure. Body weights of the animals were determined on the administration day (day 0), i.e. directly before the administration, and on the last day of the experiment. After the observation period, the animals were euthanized.

The observations of the treated skin revealed some changes in the form of barely perceptible erythema.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-11-08 to 2013-11-16

Reliability:

1 (reliable without restriction)

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:

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin Irritation:

A study was undertaken to investigate the skin irritation potential of Mn (II) IDHA in New Zealand White rabbits (OECD 404; Sornat, 2013, Report No. DDR-15/13). The study commenced with a sighting study on one animal. The powdered test item was applied in an amount of 0.5 g once to the shaved skin of one animal (rabbit No. 1) and covered with an appropriate band. The exposure lasted 4 hours. After evaluation of treated skin, in order to confirm irritation or its absence, the test item was applied to the skin of the next two animals (rabbit No. 2 and No. 3) for 4 hours in order to confirm its irritant properties. The procedure was the same as in case of rabbit No. 1. General clinical observations of the animals for morbidity and mortality were performed daily during the entire experiment. Detailed clinical observations of the treated skin were performed 1, 24, 48, and 72 hours after the end of the exposure. Body weights of the animals were determined on the administration day (day 0), i.e. directly before the administration, and on the last day of the experiment. After the observation period, the animals were euthanized.

The observations of the treated skin revealed some changes in the form of barely perceptible erythema in one animal. The reaction was fully reversible within 48 hours.

Eye irritation

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.

Justification for selection of skin irritation / corrosion endpoint:
only one study available

Justification for selection of eye irritation endpoint:
only one study available

Justification for classification or non-classification

Skin irritation

According to the classification criteria outlined in the section 3.2.2.2. (Guidance on the Application of CLP criteria, November, 2013), if a substance causes a 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 inflammation persists to the end of the observation period in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling, or in some cases where there is pronounced variability of response amoung animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above, the classification as Skin Irritant (Category 2) is assigned. The average irritation scores from gradings at 24, 48 and 72 hours were 0.3, 0.0 and 0.0 for erythema and 0.0, 0.0 and 0.0 for oedema. Based on these results, there were no positive responders. All scores were under the cut-off value (2.3) triggering classification and labelling . Therefore, Mn(II)IDHA does not meet the criteria for classification and labelling for skin irritation in accordance with European regulation (EC) No. 1272/2008.

Eye irritation

According to the classification criteria outlined in the section 3.3.2.2. (Guidance on the Application of CLP criteria, November, 2013), if a substance cause positive but reversible reactions such as corneal opacity (score ≥ 1), iritis (score ≥ 1), conjunctival redness (score ≥ 2) or conjunctival oedema (chemosis) (score ≥ 2) in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after application, the classification as Eye Irritant (Category 2) is assigned. The average irritation scores from gradings at 24, 48, and 72 hours for corneal opacity and iritis were 0.0 for three animals. Conjunctivae (erythema) mean scores of 0.7, 1.0 and 0.3 were yielded for animal 1, 2 and 3, respectively. Further, chemosis mean scores of 0.3, 0.0 and 0.0 are reported for the three tested animals, respectively. Based on these results, there were no positive responders regarding corneal opacity, iritis and conjunctivae (erythema and chemosis). All scores were under the cut-off values triggering classification and labelling. Therefore, the substance does not need to be classified and labelled as eye irritant according to Regulation (EC) No. 1272/2008.