Registration Dossier

Administrative data

Description of key information

- Skin Irritation, OECD 404, 3 New Zealand White rabbits, semiocclusive, all scores are 0.0 from gradings at 24, 48 and 72 h time points. Conclusion: not irritating.
- Eye irritation: OECD 405, 3 New Zealand White rabbits, no corneal opacity, congestion in iris of one animal at 24-hour reading time point and conjunctival congestion, swelling and ocular discharge. Only one positive responder with conjunctival erythema score of 2.3. 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:
2010-11-09 to 2010-11-19
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well-documented GLP-Guideline Study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
other: SOP/T/22: “Acute skin irritation/corrosion study”
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Slovak National Accreditation Service (No. G-024)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Experimental Department of National Research Institute of Animal Production, Balice near Kraków
- Age at study initiation: 9-month-old rabbit female (rabbit No 1), 8.5-month-old male (rabbit No 2) and one 8-month-old male (rabbit No 3)
- Housing: individually in metal cages (60 x 35 x 40 cm). Each cage was equipped with label containing information on number of study protocol, start and end date of experiment (written after termination), sex and animal’s number
- 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/h
- Photoperiod (hrs dark / hrs light): 12 / 12

Study commencement: 09.11.2010.
Experiment: from 09.11.2010. to 12.11.2010. (rabbit No 1)
from 16.11.2010. to 19.11.2010. (rabbit No 2 and No 3)
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:
72 hours
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 with 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:
mean
Remarks:
for 3 animals
Time point:
other: mean at 24, 48 and 72 hours
Score:
0
Irritation parameter:
edema score
Basis:
mean
Remarks:
for 3 animals
Time point:
other: mean at 24, 48 and 72 hours
Score:
0
Irritant / corrosive response data:
No irritating effects were noted.

Table 1: Summary of Cu (II) IDHA. Acute skin irritation/corrosion on rabbits
Administered amount of test item 0.5 g
Number of rabbit 1 2 3
Sex female male male
Mortality of animals 0/1 0/1 0/1
General clinical signs none none none
Skin erythema in area of treatment none none none
Skin oedema in area of treatment none none none
Other none none none

During the entire experiment, no pathological changes were stated on treated skin of rabbits.

Table 2 - Point evaluation of acute skin irritation/corrosion on rabbits
Acute skin irritation/corrosion on rabbits
Cu (II) IDHA
Rabbit 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 0 0 0 0 0
oedema 0 0 0 0 0
3 erythema 0 0 0 0 0
oedema 0 0 0 0 0

Body weight of animals

At the start of experiment animal No 1 weighed 3.7 kg, animal No 2 – 3.5 kg and animal No 3 – 3.6 kg.

At the end of experiment animal No 1 weighed 3.6 kg, animal No 2 – 3.5 kg and animal No 3 – 3.6 kg

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
During readings no pathological changes were stated on treated skin of rabbits. The test item Cu (II) IDHA does not irritate skin of rabbits.
Executive summary:

A study was undertaken to investigate the skin irritation potential of Cu (II) IDHA in New Zealand White rabbits (OECD 404; Kropidło, 2010, Report No. DDR-15/10). A dose of 0.5 g then was applied to a shaved, unabraded site on one animal. The powdered test item was applied in amount of 0.5 g as a single dose (applied first to multilayered gauze patch with dimensions of 2.5 x 2.5 cm, moistened with few drops of water and then laid on prepared skin) to shaved skin of one animal (rabbit No 1) and covered with appropriate band. The area of skin surface treated with the test item was about 6 cm². The gauze patches were covered with PCV foil and fixed with sticking plaster. The rabbits trunk was covered with lignin and elastic bandage was used to make circular protecting band. After an exposure time of 4 hours, the band and gauze patches were taken off and the residual test item was removed using water. The animal was observed for 72 hours. After evaluation of treated skin, in order to confirm irritation or its lack, the test item was applied to skin of another two animals (rabbit No 2 and No 3) for 4 hours. The test sites were then evaluated in accordance with the method of Draize at 1 hour, 24, 48 and 72 hours.

General clinical observations for morbidity and mortality of all animals were performed daily during the entire experiment. Detailed clinical observations of treated skin were performed after 1, 24, 48 and 72 hours since the end of exposure. Body weight of animals was determined directly before administration on day of administration of test item (day 0) and on last day of experiment. During readings no pathological changes were stated on treated skin of rabbits. No mortality or effects on body weight were observed. The test item Cu (II) IDHA does not irritate skin of rabbits.

Based on the results of this study this substance is not classified as a skin irritant in accordance with EU CLP Regulation (EC) No. 1272/2008.

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:
2010-11-09 to 2010-11-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
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
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.

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 Cu(2Na)IDHA in New Zealand White rabbits (Kropidło, 2010, Report No. DDR-15/10, according to OECD 404). A dose of 0.5 g then was applied to a shaved, unabraded site on one animal. The powdered test item was applied in amount of 0.5 g as a single dose (applied first to multilayered gauze patch with dimensions of 2.5 x 2.5 cm, moistened with few drops of water and then laid on prepared skin) to shaved skin of one animal (rabbit No 1) and covered with appropriate band. The area of skin surface treated with the test item was about 6 cm². The gauze patches were covered with PCV foil and fixed with sticking plaster. The rabbits trunk was covered with lignin and elastic bandage was used to make circular protecting band. After an exposure time of 4 hours, the band and gauze patches were taken off and the residual test item was removed using water. The animal was observed for 72 hours. After evaluation of treated skin, in order to confirm irritation or its lack, the test item was applied to skin of another two animals (rabbit No 2 and No 3) for 4 hours. The test sites were then evaluated in accordance with the method of Draize at 1 hour, 24, 48 and 72 hours. General clinical observations for morbidity and mortality of all animals were performed daily during the entire experiment. Body weight of animals was determined directly before administration on day of administration of test item (day 0) and on last day of experiment.

During readings (after 24, 48 and 72 hours) no pathological changes were stated on treated skin of rabbits. The mean erythema score and the mean oedema score were 0.0 for all time points. In addition, no mortality or effects on body weight were observed.

Eye irritation:

The acute eye irritation/corrosion study was performed in order to obtain information on health risk of Cu(2Na)IDHA in eyes of three (females) New Zealand White rabbits (Kropidło, 2010; Project No. ODR – 13/10, according to OECD 405). 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.


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 among 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 all 0.0. Based on these results, there were no positive responders. All scores were under the cut-off value (2.3) triggering classification and labelling. Therefore, Cu(2Na)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 (score1), iritis (score1), conjunctival redness (score2) or conjunctival oedema (chemosis) (score2) 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 were 0.0 for three animals. Iritis mean scores of 0.33, 0.0 and 0.0 and conjunctivae (erythema) mean scores of 1.7, 1.3 and 2.3 were yielded for each of the test animals, respectively. Furthermore, chemosis mean scores of 1.0, 0.7 and 1.3 are reported for the three tested animals, respectively. Based on these results, there were no positive responders regarding corneal opacity, iritis and conjunctival chemosis. All scores were under the cut-off values triggering classification and labelling. The only one animal with conjunctival redness score of 2.3 is the positive responder. The conjunctival erythema scores of other two animals were also under the cut-off value (2.0). Therefore, the substance does not need to be classified and labelled as eye irritant according to Regulation (EC) No. 1272/2008.