Registration Dossier

Administrative data

Description of key information

Skin Irritation: Non-irritating, OECD 404, EU Method B.4, EPA OPPTS 870.2500, JMAFF 2-1-4, Zelenák 2012.
Eye Irritation: Irritant (Category 2), OECD 405, EU Method B.5 and EPA OPPTS 870.2400, Török-Bathó 2012.

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:
06 December 2011 to 09 December 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
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:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF 2-1-4
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 12 weeks old males.
- Weight at study initiation: 3120 – 3146 g
- Housing: Individually in metal wire rabbit cages.
- Diet: Commercial feed, ad libitum.
- Water: Municipal tap water, ad libitum.
- Acclimation period: 13 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.1 to 20.7 °C
- Humidity (%): 24 to 60 %
- Air changes (per hr): 15 to 20 per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light per day, from 06:00 to 18:00 hours.
Type of coverage:
occlusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
Duration of treatment / exposure:
4 hours
Observation period:
72 hours after treatment.
Number of animals:
Three
Details on study design:
TEST SITE
- Area of exposure: A 6 cm² area on the back and flank of each animal.
- Type of wrap if used: A single layer of fine medical gauze (open-weave with large holes) of approximately 5 x 5 cm was placed over the application area. The appropriate amount of test material was carefully spread over the application area (the gauze helped maintain the test item in place). Three more layers of gauze were placed over the test material. These gauze patches were kept in contact with the skin by a patch of clear plastic with a surrounding adhesive hypoallergenic plaster to ensure continued good contact between the moistened test material and the shaved skin. The entire trunks of the animals were wrapped with plastic wrap, kept in place with medical elastic tubing.

REMOVAL OF TEST SUBSTANCE
- Washing: The test material was removed with water at body temperature.
- Time after start of exposure: 4 hours.

OBSERVATIONS
- Dermal reaction: Animals were examined for signs of erythema and edema at 60 minutes and then at 24, 48 and 72 hours after patch removal. Animals were observed for a sufficient amount of time to assess reversibility of any observed effects.
- Clinical signs: Recorded daily.
- Body weights: Recorded at the beginning and at termination of the experiment.

SCORING SYSTEM: Dermal reactions were scored according to the Draize scale (1959), see Table 1.
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
8
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritant / corrosive response data:
- One hour after patch removal: Very slight erythema (score 1) was observed in two animals and very slight edema (score 1) was noted in one animal.
- 24, 48 and 72 hours after patch removal: There were no observed clinical signs noted on the skin of the treated animals.
- As no clinical signs were observed the study was terminated after the 72 hours observation.
Other effects:
- Mortality: None observed during the study.
- Clinical: No treatment related clinical signs noted.
- Bodyweights: There was no effect of treatment on body weight. Body weights measured between 3184 and 3197g at termination.

Table 2: Erythema Formation Results

Animal No.

1 Hour

24 Hours

48 Hours

72 Hours

1

0

0

0

0

2

1

0

0

0

3

1

0

0

0

Total

2

0

0

0

 

Table 3: Edema Formation Results

Animal No.

1 Hour

24 Hours

48 Hours

72 Hours

1

0

0

0

0

2

1

0

0

0

3

0

0

0

0

Total

1

0

0

0

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Very slight erythema (score 1) was observed in two animals and very slight edema (score 1) was noted in one animal one hour after application of the test material. By 24 hours all effects were fully reversible. Under the conditions of the test, the test material was determined to be non-irritating.
Executive summary:

The potential for the test material to cause skin irritation was determined in vivo, in a study conducted under GLP conditions and in line with OCED 404, EU Method B.4, EPA OPPTS 870.2500 and Japanese MAFF (2-1-4). The test material was administered to three male New Zealand White rabbits and occluded for 4 hours, after which the test material was removed by washing with water. Dermal reactions were assessed at 1, 24, 48 and 72 hours post removal. Clinical observations were recorded daily and bodyweights were measured at initiation and termination.

Very slight erythema (score 1) was observed in two animals and very slight edema (score 1) was noted in one animal one hour after application of the test material. By 24 hours all effects were fully reversible. Under the conditions of the study, the test material was determined to be non-irritating.

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:
20 December 2011 to 03 January 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
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:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Sex: Males
- Age at study initiation: Approximately 16 weeks old.
- Weight at study initiation: Ranged from 3615 to 3904 g
- Housing: Individually in metal wire cages.
- Diet: Commercial rabbit feed, ad libitum.
- Water: Municipal tap water, as libitum.
- Acclimation period: 41 days.
- Health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined prior to starting the study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3 °C.
- Humidity (%): 28 to 67 %
- Air changes (per hr): 15 to 20 air charges per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light per day, from 06:00 to 18:00 hours.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
Duration of treatment / exposure:
Single dose
Observation period (in vivo):
14 day
Number of animals or in vitro replicates:
Three
Details on study design:
APPLICATION OF THE TEST MATERIAL
- An initial test was performed using one animal. The test material was instilled into the conjunctival sac of the left eye. The eyelids were held closed for several seconds to prevent the loss of the test material. The contralateral eye served as the control. Immediately after the administration of the test material, an assessment of the initial pain reaction was made according to the six point scale shown in Table 1.
After consideration of the ocular responses produced in the first animal, one hour after the treatment of the first animal, two additional animals were treated.
All animals were sacrificed at the end of the observation period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Washing was performed on test animals.
- Time after start of exposure: 1 hour.

OBSERVATIONS
- The eyes were examined at 1, 24, 48, 72 hours and 1 and 2 weeks after treatment. The duration of the observation period was sufficient to identify reversibility or irreversibility of changes.
- Any clinical signs of toxicity or signs of ill health during the study were recorded.
- Individual body weights were measured at the beginning and end of the observation period.

SCORING SYSTEM: The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (24 April 2002) shown in Table 2.
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #1
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #2
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #3
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #3
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: Mean at 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
- Initial pain reaction: Slight initial pain was observed in all three animals, assigned a score of 2 (see Table 1).
- Control eye: The control eye of each animal was symptom-free.
- One hour after the application: conjunctival redness (score 2), discharge and chemosis (score 3) were seen in all rabbits. All animals showed corneal opacity (score 1, areas 2, 3 or 4).
- 24 hours after treatment: conjunctival redness (score 2), discharge (score 2 or 3) and chemosis (score 2) were seen in all rabbits. All animals showed corneal opacity (score 1 or 2, areas 1, 3 or 4).
- 48 hours after treatment, conjunctival redness (score 1 or 2), discharge (score 1 or 2) and chemosis (score 1) were seen in all rabbits. All animals showed corneal opacity (score 1, areas 1 or 3).
- 72 hours after treatment: conjunctival redness (score 1) was seen in all rabbits. Conjuctival discharge (score 1) and chemosis (score 1) were seen in two animals. All animals showed corneal opacity (score 1, areas 1 or 2).
- One week after treatment: conjunctival redness (score 1) was seen in two rabbits.
- Two weeks after treatment: no signs of eye irritation or other clinical signs were observed. The study was terminated after the 2-week observation as effects were fully reversible by this time.
Other effects:
- Mortality: None observed during the study.
- Bodyweights: The body weight and body weight change were considered to be normal with no indication of treatment related effect. Final weights ranged from 3799 to 4088 g, representing a weight gain of between 181 and 187 g.
- Clinical observations: There were no clinical signs observed that could be related to treatment. The general state and behaviour of animals were normal throughout the study period.

Table 3: Individual Scores for Ocular Irritation at 1 hour

Animal No.

Score of Irritation

IPR

Conjunctivae

Opacity of Cornea

Iris

Control Eye

Other Signs

R

CH

D

OD

OE

R

1

2

3

3

1

4

0

0

-

2

2

2

3

3

1

3

0

0

-

2

3

2

3

3

1

2

0

0

-

2

 

Table 4: Individual Scores for Ocular Irritation at 24 hours

Animal No.

Score of Irritation

Conjunctivae

Opacity of Cornea

Iris

Control Eye

Other Signs

R

CH

D

OD

OE

R

1

2

2

2

1

4

0

0

-

2

2

2

2

1

3

0

0

-

3

2

2

3

2

1

0

0

-

 

Table 5: Individual Scores for Ocular Irritation at 48 hours

Animal No.

Score of Irritation

Conjunctivae

Opacity of Cornea

Iris

Control Eye

Other Signs

R

CH

D

OD

OE

R

1

2

1

1

1

3

0

0

-

2

1

1

2

1

3

0

0

-

3

1

1

2

1

1

0

0

-

 

Table 6: Individual Scores for Ocular Irritation at 72 hours

Animal No.

Score of Irritation

Conjunctivae

Opacity of Cornea

Iris

Control Eye

Other Signs

R

CH

D

OD

OE

R

1

1

1

1

1

2

0

0

-

2

1

1

0

1

2

0

0

-

3

1

0

1

1

1

0

0

-

 

Table 7: Individual Scores for Ocular Irritation at 1 week

Animal No.

Score of Irritation

Conjunctivae

Opacity of Cornea

Iris

Control Eye

Other Signs

R

CH

D

OD

OE

R

1

1

0

0

0

0

0

0

-

2

1

0

0

0

0

0

0

-

3

0

0

0

0

0

0

0

-

 

Table 8: Individual Scores for Ocular Irritation at 2 weeks

Animal No.

Score of Irritation

Conjunctivae

Opacity of Cornea

Iris

Control Eye

Other Signs

R

CH

D

OD

OE

R

1

0

0

0

0

0

0

0

-

2

0

0

0

0

0

0

0

-

3

0

0

0

0

0

0

0

-

R = redness, CH = chemosis; D = discharge; OD = opacity degree of density; OE = extent of opaque area; IPR = initial pain reaction.

Interpretation of results:
other: Category 2
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the test, the test material was determined to be irritating to the eye based on corneal opacity. Application of the test material to the rabbit’s eye caused significant conjunctival and corneal effects at one hour, which were reduced at 1 week after application. All effects were fully reversible within 14 days.
Executive summary:

The potential for the test material to cause eye irritation was assessed in an in vivo irritation study in rabbits. The study was performed under GLP conditions in line with standardised guidelines OECD 405, EU Method B.5 and EPA OPPTS 870.2400. The test material was administered to three male New Zealand white rabbits, in a single application which was washed after 1 hour. Rabbits were assessed over a 14 day observation period for signs of eye irritation, clinical sign of toxicity, mortality and bodyweight gain. Application of the test material to the rabbit’s eye caused significant conjunctival and corneal effects at one hour, which were reduced 1 week after application. The study was terminated after 14 day as all signs of irritation were fully reversed. Under the conditions of the study, the test material was determined to be an eye irritant (category 2).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation

Zelenák (2012) has been provided as the key study, where the capacity for the test material to cause skin irritation was assessed in vivo. The study was performed under GLP and in accordance with OECD 404, EU Method B. 4, EPA OPPTS 870. 2500 and Japanese MAFF (2-1-4). The study was performed to a high standard and as such has been assigned a reliability score of 1 in line with the principles for assessing data quality defined in Klimisch (1997). Very slight erythema (score 1) was observed in two rabbits and very slight edema (score 1) was noted in one rabbit one hour after application of the test material. By 24 hours all effects were fully reversible. Under the conditions of the study, the test material was determined to be non-irritating.

Kiss (2012a) has been provided as supporting data, where the potential for the test material to cause skin corrosion was assessed in vitro using the EPISKIN Model. The study was performed under GLP and in accordance with OECD 431 and EU Method B.40Bis. The study was performed to a high standard and as such has been assigned a reliability score of 1 in line with the principles for assessing data quality defined in Klimisch (1997). Under the conditions of the study, exposure to the test material resulted in increased cell viability in comparison to the negative control by approximately 50 %, which was ascribed to stimulation of metabolism rate in the cells of the disks. As the mean relative cell viability was determined to be > 35 % in comparison to the negative controls the test material was predicted to be non-corrosive to the skin, supporting the findings of the key study.

Kiss (2012b) has been provided as supporting data, where the potential for the test material to cause skin irritation was assessed in vitro using the EPISKIN Model. The study was performed under GLP and in accordance with OECD 439 and EU Method B.46. The study was performed to a high standard and as such has been assigned a reliability score of 1 in line with the principles for assessing data quality defined in Klimisch (1997). Under the conditions of the study, exposure to the test material resulted in a mean relative cell viability of 97 %. Since the cell viability was determined to be > 50 % of the negative control the test material was considered to be non-irritating, supporting the findings of the key study.

Eye Irritation:

Török-Bathó (2012) has been provided as the key study, where the capacity for the test material to cause eye irritation was assessed in an in vivo study with rabbits. The study was performed under GLP and in accordance with OECD 405, EU Method B.5 and EPA OPPTS 870. 2400. The study was performed to a high standard and as such has been assigned a reliability score of 1 in line with the principles for assessing data quality defined in Klimisch (1997). Under the conditions of the study, the test material was determined to be irritating to the eye based on corneal opacity. Application of the test material to the rabbit’s eye caused significant conjunctival and corneal effects at one hour, which were reduced at 1 week after application. All effects were fully reversible within 14 days.

Kiss (2012c) has been provided as supporting data, where the potential for the test material to cause corrosion or severe irritation was assessed in vitro in isolated chicken eyes. The study was performed to a high standard and as such has been assigned a reliability score of 1 in line with the principles for assessing data quality defined in Klimisch (1997). Under the conditions of the study, exposure to the test material resulted in an overall ICE class of 2 x I and 1 x II, which corresponds to a classification of non-irritating in accordance with the OECD guideline 438 and EU Method B. 48. However this test method does not take into account all ocular effects and the results only suggest that the test material may not cause corrosion or severe irritation. Further in vivo testing was required due to the limitations of the test method and since and negative result was obtained.


Justification for selection of skin irritation / corrosion endpoint:
The key study was performed in vivo, in line with GLP and OECD guidelines. It was assigned a reliability score of 1 in accordance with the criteria outlined in Klimisch (1997). It was therefore considered suitable to be the key study for this endpoint.

Justification for selection of eye irritation endpoint:
The key study was performed in vivo, in line with GLP and OECD guidelines. It was assigned a reliability score of 1 in accordance with the criteria outlined in Klimisch (1997). It was therefore considered suitable to be the key study for this endpoint.

Effects on eye irritation: irritating

Justification for classification or non-classification

Skin Irritation:

According to the criteria outlined in Regulation (EC) No. 1272/2008, the test material does not meet the criteria for classification as a skin irritant.

Eye Irritation:

According to the criteria outlined in Regulation (EC) No. 1272/2008 the test material meets the criteria for classification as a category 2 eye irritant with the resultant hazard phrase, H319: Causes serious eye irritation.