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

Eye irritation

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

939-867-3

Cas Number:

666748-58-9

Molecular formula:

MnC8H7NO8Na2

IUPAC Name:

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

Constituent 2

Reference substance name:

Manganese chelate of sodium salt N-[1,2 dicarboxyethyl] D,L aspartic acid

IUPAC Name:

Manganese chelate of sodium salt N-[1,2 dicarboxyethyl] D,L aspartic acid

Constituent 3

Reference substance name:

Mn (II) IDHA

IUPAC Name:

Mn (II) IDHA

Test material form:

other: aqueous solution

Details on test material:

- Name of test material (as cited in study report): Mn (II) IDHA
- Substance type: chelate
- Physical state: solid (white granules)
- Analytical purity: 9.07 % of Mn+2
- Composition of test material, percentage of components: 56.7% of MnIDHA, 40% of Na2SO4, and 3% of H2O (humidity)
- Purity test date: 2013-07-16
- Lot/batch No.: 7/2013, produced in June 2013; delivered by Sponsor on 8 July, 2013
- Expiration date of the lot/batch: June 2016.
- Stability under test conditions: June 2016.
- Storage condition of test material: at temperature between 0 – 30 °C
- Other: Data concerning the identity, purity, and stability of the test item are the responsibility of the Sponsor

Test animals / tissue source

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

Test system

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

Results and discussion

In vivo

Results

Results 1

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

Results 2

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

Results 3

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

Results 4

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

Results 5

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

Results 6

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

Results 7

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Results 8

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.

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.