Copper(2+), bis[N-[amino(imino-.kappa.N)methyl]urea-.kappa.O]-, nitrate (1:2)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

copper(2+) bis(carbamimidoylurea) dinitrate

EC Number:

800-038-5

Cas Number:

1071838-81-7

Molecular formula:

Cu(C2H6N4O)2 (NO3)2

IUPAC Name:

copper(2+) bis(carbamimidoylurea) dinitrate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: CEGAV, Argenvilliers, France
- Age at study initiation: 2 to 4 months
- Weight at study initiation: mean body weight of 3358 g (range: 2710 g to 4005 g)
- Housing: The animals were individually housed in noryl cages (Tecniplast, 65.3 cm x 65.3 cm x 45 cm). The cages were suspended in batteries over trays with absorbant paper and each cage contained a platform and enrichment (dumbbell).
- Diet (e.g. ad libitum): All animals had free access to pelleted breeding diet "type 110C", batch No. 12102, (SAFE, Augy, France)
- Water (e.g. ad libitum): All animals had free access to tap water (filtered using a 0.22 µm filter) contained in bottles.
- Acclimation period: the animals were acclimated to the study conditions for a period of at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: From: 2012-12-11 To: 2012-12-20

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 g/animal
The full amount of 0.1 g was not administered to animal A30381 because its left conjunctival sac was not large enough to receive all the test item

Duration of treatment / exposure:

1 hour

Observation period (in vivo):

evaluation of ocular reaction: 1 hour, 24, 48 and 72 hours.
Since animal A30381 showed persistent ocular reactions at the 72-hour reading, the evaluation of ocular reactions was extended up to day 10.

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Just after the 1-hour scoring, as the test item was not evacuated from the eye by the animal’s own physiological mechanisms; the eye was rinsed to remove any residual solid test item. Both eyes were rinsed with sterile isotonic saline solution (0.9% NaCl), under a volume and velocity of flow which not caused injury to the eyes.
- Time after start of exposure: Just after the 1-hour scoring

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

Redness (refers to the most severe reading for the palpebral and bulbar conjunctivae, excluding the cornea and iris)
0: blood vessels normal
1: some blood vessels hyperemic (injected)
2: diffuse, crimson color, individual vessels not easily discernible
3: diffuse, beefy red

Iris lesions
0: normal
1: markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia or injection; any or a combination of any there findings, but iris still reacting to light (a sluggish reaction being considered positive)
2: no reaction to light, haemorrhage, gross destruction (any or all of these)

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

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

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein
From the 24-hour scoring, the presence of corneal opacification was examined under a UV lamp after instillation of one or two drops of 0.5% sodium fluorescein solution (a clear fluorescence is visible in the areas of opacification). As findings were noted at 24 hours, a fluoresceine examination was conducted at each subsequent scoring time-point until a negative response was obtained.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

rabbit A30381

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not specified

Remarks:

animal A30381 was prematurely killed on day 10 of its observation period before the end of the ocular reactions reversibility

Results 2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

rabbit A30381

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not specified

Remarks:

animal A30381 was prematurely killed on day 10 of its observation period before the end of the ocular reactions reversibility

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

rabbit A30381

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

not specified

Remarks:

animal A30381 was prematurely killed on day 10 of its observation period before the end of the ocular reactions reversibility

Results 4

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

rabbit A30381

Time point:

24/48/72 h

Score:

>= 2 - <= 3

Max. score:

3

Reversibility:

not specified

Remarks:

animal A30381 was prematurely killed on day 10 of its observation period before the end of the ocular reactions reversibility

Remarks on result:

other: mean 2.7

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

rabbit A30382

Time point:

24/48 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks:

Animal A30382 was sacrificed on day 3 of the observation period for ethical reasons (serious eye damage)

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

rabbit A30382

Time point:

24/48 h

Score:

1

Max. score:

1

Reversibility:

not specified

Remarks:

Animal A30382 was sacrificed on day 3 of the observation period for ethical reasons (serious eye damage)

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

rabbit A30382

Time point:

24/48 h

Score:

3

Max. score:

3

Reversibility:

not specified

Remarks:

Animal A30382 was sacrificed on day 3 of the observation period for ethical reasons (serious eye damage)

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

rabbit A30382

Time point:

24/48 h

Score:

>= 3 - <= 4

Max. score:

4

Reversibility:

not specified

Remarks:

Animal A30382 was sacrificed on day 3 of the observation period for ethical reasons (serious eye damage)

Irritant / corrosive response data:

In the left treated eye of animal A30381, marked chemosis and severe redness of the conjunctiva were observed from day 1 to day 5. Then, moderate chemosis and redness persisted up to day 7. Slight conjunctiva redness was still observed until day 9.
Iris lesions were noted from day 1 until premature sacrifice on day 10.
Marked corneal opacity was recorded on days 2 and 3; moderate then slight corneal opacity persisted up to premature sacrifice on day 10.
In addition, neovascularisation was noted from day 2 to day 9.
In the left treated eye of animal A30382, marked to severe chemosis, severe redness of the conjunctiva and iris lesions were observed from day 1 until sacrifice on day 3.
A severe opacity was observed on almost the whole area of the cornea on days 2 and 3.
In addition, neovascularisation and lacrimation were noted from day 1 or 2, respectively.
As severe corneal opacity was observed for 2 consecutive days on almost the whole area of this animal, no mean scores were calculated and the test item was considered as inducing serious eye damage

Other effects:

No clinical signs indicative of systemic toxicity were observed in any animals.
The body weight of the animals was unaffected by the test item treatment.
Animal A30382, showing a severe corneal opacity intensity persisting for 48 hours, was humanely sacrificed (see § Pathology) on the third day of the observation period and discarded without macroscopic post-mortem examination.
Therefore decision was also taken to sacrifice animal A30381 on the same day (i.e. on day 10 of its observation period). Due to severity of lesions noted in animal A30382, it was not appropriate to observe the reversibility of ocular reactions for animal A30381.

Any other information on results incl. tables

No ocular reactions were observed in the right untreated control eye.

As the test item is a blue powder, bluish discoloration was noted in the left treated eye of both animals on day 1. Neovascularisation was noted for rabbit A30381 from day 2 to day 9 and for rabbit A30382 from day 1. Lacrimation were noted for rabbit A30382 from day 2. The body weight of the animals was unaffected by the test item treatment. No clinical signs indicative of systemic toxicity were observed in any animals

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions of this study, the test item, Copper Guanylurea Nitrate (CuGUN) (batch No. 40) was severely irritant when administered by ocular route to rabbits.
According to the criteria of CLP Regulation, the test item should be classified category 1 and assigned the signal word "danger" and the hazard statement "H318: causes serious eye damage".

Executive summary:

The objective of this study was to evaluate the potential irritant properties of the test item for the eye, Copper Guanylurea Nitrate (CuGUN), following a single administration to rabbits.

 

Methods

The test item, Copper Guanylurea Nitrate (CuGUN)(batch No. 40),was first administered to a single male New Zealand White rabbit (animal A30381).

As mean value from grading at 24, 48 and 72 hours after instillation was ≥ 2 for conjunctival edema (chemosis) and for conjunctival redness, ≥ 1 for iris lesions and for corneal opacity, the test item was administered in the left eye of a second animal (animal A30382).

 After administration to animal A30382, asserious eye damage was observed, no other animal was treated.

 The test item was administered inthe conjunctival sac of the left eye. The right eye remained untreated and served as control.

A quantity of 0.1 g/animal was used.

Just after the 1-hour scoring, both eyes were rinsed with a sterile isotonic saline solution (0.9% NaCl).

 Each animal was observed once a day for mortality and clinical signs.Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration and then daily until the sacrifice of animals, when applicable. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for animal A30381 only. Body weight was recorded on the day of treatment and at sacrifice.

At sacrifice, the animals were discarded without macroscopicpost‑mortemexamination.

 

Results

Animal A30382 was sacrificed on day 3 of the observation period for ethical reasons (serious eye damage).

Therefore, animal A30381 was prematurely killed on day 10 of its observation period before the end of the ocular reactions reversibility.

 No clinical signs indicative of systemic toxicity were observed in any animals.

 The body weight of the animals was unaffected by the test item treatment.

 In the left treated eye of animal A30381, marked chemosis and severe redness of the conjunctiva were observed from day 1 to day 5. Then, moderate chemosis and redness persisted up to day 7. Slight conjunctiva redness was still observed until day 9.

Iris lesions were noted from day 1 until premature sacrifice on day 10.

Marked corneal opacity was recorded on days 2 and 3; moderate then slight corneal opacity persisted up to premature sacrifice on day 10.

In addition, neovascularisation was noted from day 2 to day 9.

Mean scores calculated for animal A30381 over 24, 48 and 72 hours were as follows:

.         chemosis : 3.0; showing eye irritation,

.         redness of the conjunctiva: 3.0; showing eye irritation,

.         iris lesions: 1.0; showing eye irritation,

.         corneal opacity: 2.7; showing eye irritation.

In the left treated eye of animal A30382, marked to severe chemosis, severe redness of the conjunctiva and iris lesions were observed from day 1 until sacrifice on day 3.

A severe opacity was observed on almost the whole area of the cornea on days 2 and 3.

In addition, neovascularisation and lacrimation were noted from day 1 or 2, respectively.

As severe corneal opacity was observed for 2 consecutive days on almost the whole area of this animal, no mean scores were calculated and the test item was considered as inducing serious eye damage.

 

Conclusion

 Under the experimental conditions of this study, the test item, Copper Guanylurea Nitrate (CuGUN)(batch No. 40)was severely irritant when administered by ocular route to rabbits.

 According to the criteria of CLP Regulation,the test item should be classified category 1 and assigned the signal word"danger"and the hazard statement"H318: causes serious eye damage".