Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010-11-09 to 2010-11-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
other: SOP/T/23: “Acute eye irritation/corrosion study
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Slovak National Accreditation Service; Statement of GLP compliance No. G-024

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Cu (II) IDHA
- Substance type: chelate
- Physical state: solid (water soluble blue granules)
- Analytical purity: 9.82 % of Cu2+
- Purity test date: 2010-11-02
- Lot/batch No.: 1/10 (date of production: April, 2010)
- Stability under test conditions: in tightly closed packed at temperature from -10°C to +30°C
- Storage condition of test material: three years since production date
- Other: Data identifying test item concerning identity, purity and stability of test item are responsibility of Sponsor.

Test animals / tissue source

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)

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): 0.084 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

Results and discussion

In vivo

Resultsopen allclose all
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.

Any other information on results incl. tables

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

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 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.