Registration Dossier

Administrative data

Description of key information

Skin irritation: irritating (OECD 404, GLP, K, Rel.1)

Eye irritation: not irritating (OECD 405, GLP, K, Rel.1)

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:
25 September to 09 October 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 404 without any deviation.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted 24 April 2002
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on July 10, 2012/ signed on November 30, 2012)
Species:
rabbit
Strain:
New Zealand White
Remarks:
Hsdlf:NZW
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.20-2.65 kg
- Housing: Animals were individually housed in suspended cages.
- Diet: Food (2930C Teklad Global Certified Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK), ad libitum
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: 15 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: 25 September to 09 October 2012
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration: For the purpose of the study the test item was used as supplied.
Duration of treatment / exposure:
4 h
Observation period:
1, 24, 48 & 72 h and 7 and 14 days after the removal of the patch
Number of animals:
3 males
Details on study design:
PRETRAEATMENT
- On the day before the test each of a group of three rabbits was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.

TEST SITE
- Area of exposure: On the day of the test a suitable test site was selected on the back of each rabbit.
- Type of wrap if used: A quantity of 0.5 mL of the test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in position with a strip of surgical adhesive tape. To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Four hours after application the corset and patches were removed from each animal and any residual test item removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
- Immediately following removal of the patches and approximately 1, 24, 48 & 72 h and 7 and 14 days after the removal of the patch

SCORING SYSTEM:
- Method of calculation: Test sites were examined for evidence of primary irritation and scored according to the Draize scale, as described in OECD Guideline No. 404

OTHERS:
- Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks on result:
positive indication of irritation
Remarks:
due to the persistence of the reaction
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
probability of mild irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
probability of mild irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.7
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of mild irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks on result:
positive indication of irritation
Remarks:
due to the persistence of the reaction
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of mild irritation
Irritant / corrosive response data:
- Very slight erythema and very slight or slight oedema was noted at all treated skin sites immediately after patch removal. Very slight erythema with or without very slight oedema was noted at all treated skin sites one hour after patch removal. Well-defined erythema and very slight or slight oedema was noted at all treated skin sites at the 24 hour observation with well-defined erythema and slight oedema noted at the 48 and 72 hour observations. Very slight erythema and very slight oedema was noted at one treated skin site at the 7-Day observation.
- Loss of skin elasticity was noted at all treated skin sites at the 48 and 72 hour observations. Moderate desquamation was noted at all treated skin sites at the 7-Day observation. Small superficial scattered scabs and hardened light brown coloured scab were noted at two treated skin sites at the 7-Day observation. Scab lifting to reveal glossy skin was also noted at one treated skin site at the 7-Day observation. Adverse reactions prevented evaluation of erythema at one treated skin site at the 7-Day observation. Glossy skin was noted at two treated skin sites at the 14-Day observation.
- One treated skin site appeared normal at the 14-Day observation.
Other effects:
All animals showed expected gain in bodyweight during the study.

Table 7.3.1/1: Mean irritant/corrosive response data for each animal at each observation time

Score at time point

Erythema (Animal no 1 / 2 / 3)

Max. score 4

Oedema (Animal no 1 / 2 / 3)

Max. score 4

Immediately

1 / 1 / 1

2 / 1 / 2

1 h

1 / 1 / 1

1 / 0/ 1

24 h

2 / 2 / 2

2 / 1 / 2

48 h

2 Le / 2 Le / 2 Le

2 / 2 / 2

72 h

2 Le / 2 Le / 2 Le

2 / 2 / 2

7 days

1SsSpD / 0 D /?eSsSpSgD

1 / 0 / 0

14 days

0G / 0 /0G

0 / 0 / 0

Mean 24, 48 and 72 h

2.0 / 2.0 / 2.0

2.0 / 1.7 / 2.0

Le = Loss of skin elasticity

Ss = Small superficial scattered scabs

Sp = Hardened light brown coloured scab

Sg = Scab lifting to reveal glossy skin

D = Moderate desquamation

G = Glossy skin

?e = Adverse reactions prevent evaluation of erythema

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Under the experimental conditions of this study, the test item is classified as irritant (Category 2) according to Regulation (EC) No 1272/2008 (CLP) and to GHS based on the persistence of inflammation until the end of the 14 -day observation period in 2 out of 3 animals.
Executive summary:

In a dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 0.5 mL of test item was applied on the clipped skin of the dorsal/flank area (back) of 3 male New Zealand White rabbits. Test sites were covered with a semi-occlusive dressing for 4 h. Skin irritation was assessed and scored according to the Draize scale at 1, 24, 48, 72 h and 7 and 14 days after the removal of the patch.

Very slight erythema and very slight or slight oedema was noted at all treated skin sites immediately after patch removal. Very slight erythema with or without very slight oedema was noted at all treated skin sites one hour after patch removal. Well-defined erythema and very slight or slight oedema was noted at all treated skin sites at the 24 hour observation with well-defined erythema and slight oedema noted at the 48 and 72 hour observations. Very slight erythema and very slight oedema was noted at one treated skin site at the 7-Day observation. Loss of skin elasticity was noted at all treated skin sites at the 48 and 72 hour observations. Moderate desquamation was noted at all treated skin sites at the 7-Day observation. Small superficial scattered scabs and hardened light brown coloured scab were noted at two treated skin sites at the 7-Day observation. Scab lifting to reveal glossy skin was also noted at one treated skin site at the 7-Day observation. Adverse reactions prevented evaluation of erythema at one treated skin site at the 7-Day observation. Glossy skin was noted at two treated skin sites at the 14-Day observation. One treated skin site appeared normal at the 14-Day observation.

The individual scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.0 / 2.0 / 2.0 for erythema and 2.0 / 1.7 / 2.0 for oedema.

Under the experimental conditions of this study, the test item is classified as irritant (Category 2) according to Regulation (EC) No 1272/2008 (CLP) and to GHS based on the persistence of inflammation until the end of the 14 -day observation period in 2 out of 3 animals.

This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
17 October to 01 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 405 without deviation.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted 24 April 2002
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on July 10, 2012/ signed on November 30, 2012)
Species:
rabbit
Strain:
New Zealand White
Remarks:
Hsdlf:NZW
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.55-2.99 kg
- Housing: Animals were individually housed in suspended cages.
- Diet: Food (2930C Teklad Global Certified Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK), ad libitum.
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: 15 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: 17 October to 01 November 2012
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): Undiluted
Duration of treatment / exposure:
No washing was done
Observation period (in vivo):
1, 24, 48 and 72 h after instillation of test item
Number of animals or in vitro replicates:
3 males
Details on study design:
PRETREATMENT
- Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.

PROCEDURE
- Initially, a single rabbit was treated. A volume of 0.1 ml of the test item was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1.
- After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: Draize scale as described in the OECD guideline No. 405.

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

OTHERS
- Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.3
Max. score:
2
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 h
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 h
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72 h
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
- No corneal effects were noted during the study.
- Iridial inflammation was noted in one treated eye one and 24 hours after treatment.
- Moderate conjunctival irritation was noted in all treated eyes one hour after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 24 hour observation. Minimal conjunctival irritation was noted in all treated eyes at the 48 hour observation.
- All treated eyes appeared normal at the 72 hour observation.
Other effects:
All animals showed expected gain in bodyweight during the study.

Table 7.3.2/1: Eye irritation response data for each animal at each observation time

Score at time point

Cornea

Iris

(/2)

Conjunctivae

Opacity

(/4)

Area

(/4)

Redness

(/3)

Chemosis

(/4)

Discharge

(/3)

1 h

0 / 0 / 0

0 / 0 / 0

1 / 0 / 0

2 / 2 / 2

2 / 1 / 1

1 / 1 / 1

24 h

0 / 0 / 0

0 / 0 / 0

1 / 0 / 0

2 / 2 / 1

1 / 1 / 1

0 / 1 / 0

48 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

1 / 1 / 1

0 / 1 / 0

0 / 0 / 0

72 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

Mean 24, 48 and 72 h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

0.3 / 0.0 / 0.0

1.0 / 1.0 / 0.7

0.3 / 0.7 / 0.7

0.0 / 0.3 / 0.0

Reversibility

 -

 -

Completely

reversible

Completely

reversible

Completely reversible

Completely reversible

Average time (unit) for reversion

-

-

48 h

72 h

72 h

48 h

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the test item is not classified as eye irritant according to Regulation (EC) No. 1272/2008 (CLP) and to GHS.
Executive summary:

In an eye irritation study performed according to the OECD Guideline No. 405, and in compliance with GLP, 0.1 mL of undiluted test item was instilled into the right eye of 3 male New Zealand White(Hsdlf:NZW) strain rabbits. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test item, and then released. The left eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48, and 72 h after treatment and graded according to the Draize method.

 

No corneal effects were noted during the study. Iridial inflammation was noted in one treated eye one and 24 hours after treatment. Moderate conjunctival irritation was noted in all treated eyes one hour after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 24 hour observation. Minimal conjunctival irritation was noted in all treated eyes at the 48 hour observation. All treated eyes appeared normal at the 72 hour observation.

 

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.0 / 0.0 / 0.0 for cornea score; 0.3 / 0.0 / 0.0 for iris score; 1.0 / 1.0 / 0.7 for conjunctivae score and 0.3 / 0.7 / 0.7 for chemosis score.

 

Under the experimental conditions of this study, the test item is not classified as eye irritant according to Regulation (EC) No. 1272/2008 (CLP) and to GHS.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

A key study was identified (Harlan, 2012). In this dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, a single 4 -Hour, semi-occluded application of the test item to the intact skin of three male rabbits produced well-defined erythema and slight oedema. Other skin reactions noted were loss of skin elasticity, scabbing, moderate desquamation and glossy skin. One treated skin site appeared normal while glossy skin persisted for two animals at the 14-Day observation.

The individual scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.0/2.0/2.0 for erythema and 2.0/1.7/2.0 for oedema. Due to high scores and persistence of the reactions, the substance is therefore classified for skin irritation.

Eye irritation:

A key study was identified (Harlan, 2012). In this eye irritation study performed according to the OECD guideline 405, and in compliance with GLP, 0.1 mL of undiluted test material was instilled into one eye of 3 male New Zealand White rabbits. The other eye remained untreated and served as control. The upper and lower eyelids were held together for about one second immediately after instillation. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h after treatment and graded according to the Draize method.

The calculated mean score for each animal within 3 scoring times (24, 48 and 72 h) were 0.0/0.0/0.0 for the corneal opacity, 0.3/0.0/0.0 for iridial inflammation, 1.0/1.0/0.7 for conjunctival redness and 0.3/0.7/0.7 for chemosis. The effects observed were all reversible within 72 hours. The substance is therefore not classified for eye irritation.

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Based on the available data, the substance should be classified as Skin irr. Category 2 (H315: Causes skin irritation) according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

No additional self-classification is proposed regarding eye irritation according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Although there are currently no validated animal tests that deal with specifically with respiratory tract irritation, no self-classification is proposed regarding this endpoint based on the absence of effects in the acute inhalation toxicity study.