Registration Dossier

Administrative data

Description of key information

SKIN
Not irritating, OECD 404, EU Method B.4, Pooles (2009)
EYE
Not irritating, OECD 405, EU Method B.5, Pooles (2010)

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:
3 November 2009 to 13 November 2009
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
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Hillcrest, Belton, Loughborough, UK
- Age at study initiation: 12 - 20 weeks old
- Weight at study initiation: 2.17 - 2.43 kg
- Housing: Individually housed in suspended cages
- Diet: 2030 Teklad Global Rabbit diet (Harlan Teklad, Blackthorn, Bicester, Oxon, UK) available ad libitum
- Water: Mains drinking water available ad libitum
- Acclimation period: Minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): A minimum of 15 changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hour cycle
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied : 0.5 g of test material sufficiently moistened in 0.5 mL of distilled water to achieve a paste.
Duration of treatment / exposure:
Animal were exposed for 4 hours
Observation period:
Animals were observed for 72 hours for skin reactions
Number of animals:
3 animals in total (1 animal used in the initial test, with a further 2 animals in the main test)
Details on study design:
TEST SITE
- Area of exposure: The test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch
- Type of wrap if used: The trunk of each rabbit was wrapped in an elasticated corset

REMOVAL OF TEST MATERIAL
- Washing: Any residual test material was removed by gentle swabbing with cotton wool soaked in distilled water
- Time after start of exposure: 4 hours post administration of test material

SCORING SYSTEM: Draize JH (1959) “Dermal Toxicity” In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Associations of Food and Drug Officials of the United States, Austin, Texas p.46-59
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 24 and 72 hours post dosing
Score:
0
Max. score:
8
Reversibility:
other: Not applicable, no effects observed during the study.
Irritant / corrosive response data:
The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the three test rabbits (12 values) and this total was divided by six to give the primary irritation index of the test material.

The test material produced a primary irritation index of 0.0.

Measurement of pH

The pH of the test material was determined prior to commencement of the study and found to be as follows:

Preparation

pH Measurement

immediately

after 10 minutes

after 20 minutes

 10 % w/w aqueous preparation of the test material

 8.9

 9.6

 9.6

Table 1: Individual Skin Reactions

Skin Reaction

Observation Time (Following Patch Removal)

Individual Scores – Rabbit Number and Sex

Total

68637 Male

68638 Male

68661 Male

Erythema/Eschar Formation

Immediately

0

0

0

(0)

1 Hour

0

0

0

(0)

24 Hours

0

0

0

0

48 Hours

0

0

0

(0)

72 Hours

0

0*

0*

0

Oedema Formation

Immediately

0

0

0

(0)

1 Hour

0

0

0

(0)

24 Hours

0

0

0

0

48 Hours

0

0

0

(0)

72 Hours

0

0*

0*

0

Sum of 24 and 72-hour readings (S): 0

Primary Irritation Index (S/6): 0/6 = 0.0

Classification: Non-Irritant

() = total values not included in calculating the primary irritation index

 

Table 2: Individual Bodyweights and Bodyweight Changes

Rabbit Number and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

68637 Male

2.17

2.21

0.04

68638 Male

2.37

2.43

0.06

68661 Male

2.43

2.52

0.09

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material produced a primary irritation index of 0.0 and was classified as a non-irritant to rabbit skin. No corrosive effects were noted.
Executive summary:

The skin irritation potential of the test material was investigated in vivo in a study which was conducted under GLP conditions and in accordance with the standardised guidelines OECD 404 and EU Method B.4.

During the study no signs of skin irritation were noted at any of the observation points. The test material was therefore concluded to be a non-irritant.

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:
9 November 2009 to 19 November 2009
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
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories U.K. Limited, Hillcrest, Belton, Loughborough, UK
- Age at study initiation: 12 - 20 weeks old
- Weight at study initiation: 2.18 - 2.39 kg
- Housing: Animals were housed individually in suspended cages
- Diet: Free access to 2030 Teklad Global Rabbit diet (Harlan Teklad, Blackthorn, Bicester, Oxon, UK)
- Water: Free access to mains drinking water
- Acclimation period: A minimum of 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): At least 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hour cycle.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied : 100 mg
Duration of treatment / exposure:
Up to 72 hours
Observation period (in vivo):
Animals were assessed up to 72 hours after exposure (assessments were made at 1, 24, 48 and 72 hours post treatment).
Number of animals or in vitro replicates:
1 animal was initially treated, two further animals were treated thereafter
Details on study design:
APPLICATION OF TEST MATERIAL:
Initially a single rabbit was treated with 100 mg of test material. The test material was administered 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 test material and then released. The left eye served as a control and remained untreated. Immediately after administration of the test material and assessment of the initial pain reaction was made. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

SCORING SYSTEM:
Assessment of ocular damage/irritation was made 1, 24, 48 and 72 hours post dosing using the scoring system from Draize JH (1977) “ Dermal and Eye Toxicity Tests” In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p. 48 to 49. Any other ocular effects were also noted at this point along with any clinical signs of toxicity if present. Individual bodyweights were recorded on day 0 and 3. The numerical values corresponding to each animal, tissue and observation time were recorded. These scores were then assessed using a modified version of the system described by Kay JH and Calandra JC (1962), J. Soc. Cosmet. Chem. 13, 281-289. If evidence of irreversible ocular damage is noted, the test material will be classified as corrosive to the eye.

TOOL USED TO ASSESS SCORE: standard ophthalmoscope
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
80
Reversibility:
other: not applicable
Remarks on result:
other: No corneal effects observed
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
10
Reversibility:
other: not applicable
Remarks on result:
other: No iridial effects observed
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
4
Max. score:
20
Reversibility:
fully reversible within: 48 hours
Irritant / corrosive response data:
The test material produced a maximum group mean score of 10.0

To calculate the maximum group mean score, encompassing all reactions, the following equations were used:
Cornea = (E x F) x 5
Where:
E = Degree of opacity
F = Area of Cornea Involved

Iris = (D x 5)

Conjunctivae = (A + B + C) x 2
Where:
A = Redness
B = Chemosis
C = Discharge

Measurement of pH

The pH of the test material was determined prior to commencement of the study and found to be as follows:

 

Preparation

pH Measurement

immediately

after 10 minutes

after 20 minutes

 10 % w/w aqueous preparation of the test material

8.9

9.6

9.6

 

 

Table 1: Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

68648 Male

68668 Male

68669 Male

IPR = 2

IPR = 2

IPR = 2

Time After Treatment

1 Hr

24 Hrs

48 Hrs

72 Hrs

1 Hr

24 Hrs

48 Hrs

72 Hrs

1 Hr

24 Hrs

48 Hrs

72 Hrs

Cornea

E = Degree of opacity

0

0

0

0

0

0

0

0

0

0

0

0

F = Area of Cornea Involved

0

0

0

0

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

0

0

0

0

Iris

D

0

0

0

0

0

0

0

0

0

0

0

0

Score (D x 5)

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctivae

A = Redness

2

1

0

0

2

1

0

0

2

1

0

0

B = Chemosis

2

1

0

0

2

1

0

0

2

1

0

0

C = Discharge

1

0

0

0

1

0

0

0

1

0

0

0

Score (A + B + C) x 2

10

4

0

0

10

4

0

0

10

4

0

0

Total Score

10

4

0

0

10

4

0

0

10

4

0

0

IPR = Initial pain reaction

 

Table 2: Individual Total Scores and Group Mean Scores for Ocular Irritation

Rabbit Number and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

68648 Male

10

4

0

0

68668 Male

10

4

0

0

68669 Male

10

4

0

0

Group Total

30

12

0

0

Group Mean Score

10.0

4.0

0.0

0.0

 

Table 3:Individual Bodyweights and Bodyweight Changes

Rabbit Number and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

68648 Male

2.33

2.37

0.04

68668 Male

2.39

2.42

0.03

68669 Male

2.18

2.30

0.12

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Slight conjunctival redness and chemosis (score 1) was observed 24 hours following administration of the test material in the treated eyes of all three animals; all treated eyes appeared normal 48 hours after administration. Under the conditions of the study the test material was determined to be not irritating to eyes.
Executive summary:

The eye irritation potential of the test material was investigated in vivo in a study which was conducted under GLP conditions and in accordance with the standardised guidelines OECD 405 and EU Method B.5.

No corneal or iridial effects was noted during the study. Moderate conjuctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24-hour observation. Under the conditions of the study the test material was determined to be not irritating to eyes.

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

Respiratory irritation

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

Additional information

Skin

In vitro testing concluded that the test material was unlikely to be irritating to skin following exposure. An in vivo test was subsequently performed to confirm findings of the in vitro study.

The skin irritation potential of the test material was investigated in vivo in a study which was conducted under GLP conditions and in accordance with the standardised guidelines OECD 404 and EU Method B.4.

During the study no signs of skin irritation were noted at any observation point. The test material was therefore concluded to be a non-irritant to the skin.

Eye

In vitro testing concluded that the test material was unlikely to be an eye irritant. An in vivo test was subsequently performed to confirm findings of the in vitro study.

The eye irritation potential of the test material was investigated in vivo in a study which was conducted under GLP conditions and in accordance with the standardised guidelines OECD 405 and EU Method B.5.

No corneal or iridial effects were noted during the study. Moderate conjuctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24-hour observation. The test material was therefore concluded to be not irritating to eyes.


Justification for selection of skin irritation / corrosion endpoint:
The key study was selected as such because it represents the only available in vivo study.

Justification for selection of eye irritation endpoint:
The key study was selected as such because it represents the only available in vivo study.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to either skin or eye irritation.

In accordance with the criteria for classification as defined in Annex VI, Directive 67/548/EEC (DSD), the substance does not require classification with respect to either skin or eye irritation.