Registration Dossier

Administrative data

Description of key information

Skin irritation: The test substance is not considered to be irritating to skin.
Eye irritation: The test substance is not considered to be irritating to the 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:
31 August 1995- 07 February 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
other: EEC Guidelines, Annex V Methods (Directive 92/69/EEC of 31 July 1992)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Olac Limited, Shaw's Farm, Blackthorn, Bicester, OX6 0TP
- Age at study initiation: yound adult
- Weight at study initiation: 3.36- 3.56 kg
- Housing: the rabbits were housed individually in aluminium cages (48 x 61 x 46 cm) with grid floors beneath which were absorbent paper lined trays. Floors, cage racks, walls and ceilings were cleaned with disinfectant solution (0.5 % Tego 2000, Th Goldschmidt and Company Limited) as necessary.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-20
- Humidity (%): 61
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/ 12

IN-LIFE DATES: From: 28 September 1995 To: 12 October 1995
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Concentration (if solution): 0.5 mL
Duration of treatment / exposure:
4 h
Observation period:
1, 24, 48, 72 h continuing up to 14 d
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 2.5 cm x 2.5 cm
- % coverage:
- Type of wrap if used: The patch of gauze was covered with Micropore tape and the trunk was loosely bound with Elastoplast Elastic Bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): skin wiped with water
- Time after start of exposure: 4 h

SCORING SYSTEM
Erythema and Eschar Formation:
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4

Oedema formation:
No oedema: 0
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well defined by definite raising): 2
Moderate oedema (area raised approximately 1 mm): 3
Severe oedema (area raised by more than 1 mm and extending beyond area of exposure): 4
Irritation parameter:
erythema score
Basis:
mean
Remarks:
Animal 1
Time point:
other: 24 h, 48 h, 72 h
Score:
1
Irritation parameter:
erythema score
Basis:
mean
Remarks:
Animal 2
Time point:
other: 24 h, 48 h, 72 h
Score:
1.3
Irritation parameter:
erythema score
Basis:
mean
Remarks:
Animal 3
Time point:
other: 24 h, 48 h, 72 h
Score:
1
Irritation parameter:
edema score
Basis:
mean
Remarks:
Animal 1
Time point:
other: 24 h, 48 h, 72 h
Score:
0
Irritation parameter:
edema score
Basis:
mean
Remarks:
Animal 2
Time point:
other: 24 h, 48 h, 72 h
Score:
0
Irritation parameter:
edema score
Basis:
mean
Remarks:
Animal 3
Time point:
other: 24 h, 48 h, 72 h
Score:
0
Irritant / corrosive response data:
All animals showed slight erythema from 1- 72 h after patch removal. In 1 animal an increase to more definite erythema was noted at 24 h after patch removal. This animal continued to show slight erythema for 14 d after patch removal, with the skin becoming dry and flaky at the test site after 13 d.

No oedema was noted during the observation period.

Hexenyl acetate cis and trans acute dermal irritation test in rabbits reaction scores

Rabbit

Erythema

1 h

24 h

48 h

72 h

4 d

7 d

10 d

11 d

13 d

14 d

301

1

1

1

1

0

0

0

0

-

-

302

1

2

1

1

1

1

1

1

1*

1*

303

1

1

1

1

0

0

0

0

-

-

 

Rabbit

Oedema

1 h

24 h

48 h

72 h

4 d

7 d

10 d

11 d

13 d

14 d

301

0

0

0

0

0

0

0

0

-

-

302

0

0

0

0

0

0

0

0

0

0

303

0

0

0

0

0

0

0

0

-

-

Animals were shaved at 11 days after patch removal

h = hours after patch removal

d = days after patch removal

* = skin dry and flaky at test site

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance was assessed for acute dermal irritation according to OECD guideline 404. The test substance is classifed as not irritating although it was found to be a mild irritant to rabbit skin under the conditions of this test.
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:
The study was performed between 30 April 2012 and 11 May 2012
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)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
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 UK Ltd, Leicestershire, UK
- Age at study initiation: 12-20 weeks old
- Weight at study initiation: 2.45 or 2.94 kg
- Housing: the animals were individually housed in suspended cages. The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a lvel that might have affected the purpose or integrity of the study.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23
- Humidity (%): 30 to 70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: The left eye remained untreated and was used for control purposes
Amount / concentration applied:
0.1 mL
Observation period (in vivo):
Immediately and 1, 24, 48 and 72 h
Number of animals or in vitro replicates:
2
Details on study design:
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 1 sec immediately after treatment, to prevent loss of the test item, and then released. Immediately after administration of the test item, an assessment of the initial pain reaction was made.

After consideration of the ocular responses produced in the first treated animal, a second animal was treated.

Any other ocular effects were also noted. Any clinical signs of toxicity, if present, were also recorded. Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.


SCORING SYSTEM: Draize scale (see Appendix 2 attached). If evidence of irreversible ocular damage is noted, the test item will be classified as corrosive to the eye.

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Irritation parameter:
cornea opacity score
Basis:
other: mean score for all animals
Time point:
other: 24, 48, 72 h
Score:
0
Irritation parameter:
iris score
Basis:
other: mean score for all animals
Time point:
other: 24, 48, 72 h
Score:
0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
other: mean score for animal 1
Time point:
other: 24, 48, 72 h
Score:
1
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
other: mean score for animal 2
Time point:
other: 24, 48, 72 h
Score:
0.67
Irritation parameter:
chemosis score
Basis:
other: mean score for animal 1
Time point:
other: 24, 48, 72 h
Score:
0.67
Irritation parameter:
chemosis score
Basis:
other: mean score for animal 2
Time point:
other: 24, 48, 72 h
Score:
0.33
Irritant / corrosive response data:
No corneal effects were noted during the study.

Iridial inflammation was noted in 1 treated eye 1 h after treatment.

Moderate conjunctival irritation was noted in both treated eyes 1 h after treatment. Moderate conjunctival irritation persisted in 1 treated eye with minimal conjunctival irritation noted in the other treated eye at the 24-Hour observation. Minimal conjunctival irritation was noted in both treated eyes at the 48-Hour observation.

Both treated eyes appeared normal at the 72-Hour observation.
Other effects:
Bodyweight: both animals showed expected gain in bodyweight during the study.

Individual scores and individual total scores for ocular irritation

Rabbit number and sex

71969 Male

71992 Male

IPR= 2

IPR= 2

Time after treatment

1 Hour

24 Hours

48 Hours

72 Hours

1 Hour

24 Hours

48 Hours

72 Hours

CORNEA

E= Degree of opacity

0

0

0

0

0

0

0

0

F= Area of cornea involved

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

IRIS

D

1

0

0

0

0

0

0

0

Score (D x 5)

5

0

0

0

0

0

0

0

CONJUNCTIVAE

A= Redness

2

2

1

0

2

1

1

0

B= Chemosis

2

1

1

0

1

1

0

0

C= Discharge

2

1

0

0

1

0

0

0

Score (A + B + C) x 2

12

8

4

0

8

4

2

0

Total score

17

8

4

0

8

4

2

0

 IPR= initial pain reaction

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

71969 Male

17

8

4

0

71992 Male

8

4

2

0

Group total

25

12

6

0

Group mean score

12.5

6.0

3.0

0.0

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance was assessed for acute eye irritation according to OECD guideline 405. The test item is classified as not irritating although it was found to be a mild irritant under the conditions of this study according to a modified Kay and Calandra classification system.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

Skin irritation: The test material was applied as supplied to the shaved dorsal area of the trunk of each rabbit under a semi-occlusive patch which remained in position for 4 h. Skin reactions were then assessed 1, 24, 48 and 72 h continuing up to 14 d after patch removal. All animals showed slight erythema from 1- 72 h after patch removal. In 1 animal more defined erythema was noted at 24 h after patch removal. This animal continued to show slight erythema up to 14 d after patch removal. No oedema was noted during the observation period. The test substance is classified as not irritating although it was found to be a mild irritant to rabbit skin under the conditions of this test.

Eye irritation: The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White Rabbit. The method was designed to be compatible with OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/ Corrosion” (adopted 24 April 2002) and Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008. A single application of the test item to the non-irrigated eye of 2 rabbits produced iridial inflammation and moderate conjunctival irritation. Both treated eyes appeared normal at the 72-Hour observation. The test item is classified as not irritating although it was found to be a mild irritant under the conditions of this study according to a modified Kay and Calandra classification system.


Justification for selection of skin irritation / corrosion endpoint:
The study is a GLP guideline study and is well documented and has been assigned a reliability 1.

Justification for selection of eye irritation endpoint:
The study is a GLP guideline study and is well documented and has been assigned a reliability 1.

Justification for classification or non-classification

Skin irritation: The available study has been assigned reliability 1 and is considered as acceptable for classification. The test material did not produce a mean value of ≥2, 3- ≤4 for erythema or edema in 2 of the 3 animals from gradings at 24 and 72 h after patch removal. Therefore, the test item can be considered to be non-classified.

Eye irritation: The available study has been assigned reliability 1 and is considered as acceptable for classification. The test item did not produce a mean value of ≥1 for corneal opacity and/or ≥1 for iritis and/or ≥2 conjunctival redness and/or ≥2 conjunctival oedema (chemosis), calculated as a mean over 24, 48 and 72 h, for the animals tested. Therefore, the test item can be considered to be non-classified.