Registration Dossier

Administrative data

Description of key information

Skin irritation (OECD 404), rabbit: irritating

Eye irritation (OECD 405): not irritating

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:
from 1994-10-18 to 1994-12-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study performed under GLP without deviations
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
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, D-97633 Sulzfeld.
- Age at study initiation: not reported
- Weight at study initiation: 2.0 - 2.5 kg
- Housing: individual caging in metal wire cages
- Diet (e.g. ad libitum): Altromin 2023 diet for rabbits, ad libitum
- Water (e.g. ad libitum): tap water from an automatic watering system, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5
- Humidity (%): 50 %
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 1994-10-18
Type of coverage:
semiocclusive
Preparation of test site:
other: Hair was clipped on the dorsal area of the trunk one day before the application
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
Duration of treatment / exposure:
4 hours
Observation period:
1, 24, 48 and 72 hours after patch removal. Additional examinations 6, 8 and 10 days after patch removal.
Number of animals:
3 females
Details on study design:
TEST SITE
- Area of exposure: median on the dorsal thoracal region
- % coverage: test substance spread over an area of approximately 6 cm2
- Type of wrap if used: plastic collar

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test substance was wiped off using wet cellulose tissue
- Time after start of exposure: 4 hours

SCORING SYSTEM:
ERYTHEMA/ESCHAR FORMATION
0: No erythema
1: Very slight erythema (barely perceptible)
2: Well-defined erythema
3: Moderate to severe erythema
4: Severe erythema (beet redness) or eschar formation (injuries in depth)

OEDEMA FORMATION
0: No oedema
1: Very slight oedema (barely perceptible)
2: Slight oedema (edges of area well defined by definite raising)
3: Moderate oedema (raised approximately 1 mm)
4: Severe oedema (raised more than 1 mm and extending beyond area of exposure)
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
3.3
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
3.7
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
3.7
Max. score:
4
Reversibility:
fully reversible within: 10 days
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 10 days
Other effects:
Animal 1: Eschar formation after 72 hours. Eschar almost completely desquamated after 6 days and completely desquamated after 8 days, with intact skin below.
Animal 2: No additional observations.
Animal 3: Eschar on four sites, diameter approximately 1 mm each from 48 hours to 8 days. On day 8, eschar completely desquamated, intact skin below.

Scores of the application sites

Time after the end of exposure

Erythema / Eschar formation

Oedema formation

Animal No

1041

1042

1043

1041

1042

1043

1 h

2

2

2

3

3

3

24 h

2

2

3

3

2

4

48 h

4

2

4

4

2

4

72h

4

2

4

4

2

4

mean(24-72h)

3.3

2.0

3.7

3.7

2.0

4.0

6 d

2

1

4

2

1

4

8 d

0

0

1

0

0

2

10 d

 

 

0

 

 

0

Interpretation of results:
other: EU CLP criteria
Remarks:
Skin irritating (Category 2)
Conclusions:
The substance is irritating to the skin.
Executive summary:

The potential of the substance to induce skin irritation was investigated in a GLP study in conformance with OECD TG 404.

0.5 mL test substance was applied to the intact skin of 3 rabbits in a semi-occlusive application. After 4 hours, residual test substance was wiped off. Animals were examined for erythema/eschar and oedema as well as for other local and systemic signs approximately 1, 24, 48 and 72 hours after patch removal. Additional examinations were performed 6, 8 and 10 days after patch removal. The mean of the erythema/eschar scores at 24, 48 and 72 hours was 3.3, 2.0 and 3.7 in animal 1, 2 and 3, respectively.

The mean of the oedema scores at 24, 48 and 72 hours was 3.7, 2.0 and 4.0. Scores returned to "0", i. e. normal, within 8 days after patch removal in 2/3 animals respectively within 10 days after the end of exposure in the third animal.

In conclusion, the substance has to be regarded as irritating to the skin.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study performed under GLP without deviations
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
Species:
rabbit
Strain:
other: SPF albino rabbits of the stock Mol:Russian
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Mollegaard Breeding and Research Centre Ltd., Ejby, DK-4623 Lille Skensved
- Age at study initiation: not reported
- Weight at study initiation: 2.5 - 2.6 kg
- Housing: caged single in PPL cages with perforated floor
- Diet (e.g. ad libitum): free access to feed pellets
- Water (e.g. ad libitum): free access to bottles with domestic quality drinking water acidified with hydrochloric acid to pH 2.5 in order to prevent microbial growth
- Acclimation period: at least 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 3
- Humidity (%): 55 ± 15
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: The right eye remained untreated and served as control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
single application
Observation period (in vivo):
The eyes were examined and the grade of ocular reaction was recorded 1 and 24 hours after application. After the first 24 hour reading, Fluoreszein was instilled. After rinsing with 20 mL 0.9% sodium chloride solution, the eyes were examined again using UV-light to detect possible corneal damage. The eyes were also examined 48 and 72 hours as well as on day 7 after the treatment.
Number of animals or in vitro replicates:
3 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM:
CORNEA:
Opacity degree of density (Area most dense taken for reading)
0: No ulceration or opacity
1: Scattered or diccuse area of opacity (other than slight dulling of normal luster), details of iris clearly visible.
2: Easily discernible translucent areas, details of iris slightly obscured.
3: Nacreous areas, no details of iris visible, size of pupil barely discernible
4: Complete corneal opacity, iris not discernible

Area of cornea involved
1: One quarter (or less) but not 0
2: More than 1 quarter, but less than half
3: More than half, but less than 3 quarters
4: More than 3 quarters, up to whole area

IRIS
0: Normal
1: Markedly deepened folds, congestion, swelling, moderate circumcorneal injection (any of these or combination of any thereof), iris still reacts to light (sluggish reaction is positive)
2: No reaction to light, haemorrhage, gross destruction (any or all of these)

CONJUNCTIVA
Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
0: vessels normal
1: Some vessels definitely injected
2: Diffuse, crimson red, individual vessels not easily discernible
3: Diffuse beefy red

Chemosis
0: No swelling
1: Any swelling above normal (includes nictitating membrane)
2: Obvious swelling with partial eversion of lids
3: Swelling with lids about half closed
4: Swelling with lids more than half closed

Discharge
0: No discharge
1: Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs, and considerable area around the eye

TOOL USED TO ASSESS SCORE: hand held inspection lamp fitted with white and UV-light with 2 x magnification, fluorescein (after first 24 hour reading)
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
mean
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Redness
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
0.8
Max. score:
4
Reversibility:
fully reversible within: 7 days
Interpretation of results:
other: EU CLP criteria
Remarks:
not classified
Conclusions:
The substance was not irritating to the eye under the conditions of the test.
Executive summary:

The primary eye irritant effect of the test substance was investigated according to OECD TG 405.

Three female albino rabbits were exposed to 0.1 mL of the test article in the left eye. The eyes were examined and the changes were graded according to a numerical scale 1, 24, 48, 72 and on day 7 hours after dosing. After the treatment, only slight changes were observed in the eyes. The following mean values, based on the results from the 24, 48 and 72 hour readings, were calculated: Cornea opacity: 0/4, iris lesion: 0/2, redness of conjunctiva: 1/3, odema of conjunctiva: 0.8/4.

It is concluded that the test substance caused no eye irritation under the conditions of the test.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

The potential of the registration item to induce skin irritation was investigated in a GLP study in accordance with OECD TG 404 (1994). 0.5 mL test substance was applied to the intact skin of each of 3 rabbits in a semi-occlusive application. After 4 hours, residual test substance was wiped off. Animals were examined for erythema/eschar and edema as well as for other local and systemic signs approximately 1, 24, 48 and 72 hours after patch removal. Additional examinations were performed 6, 8 and 10 days after patch removal. The mean of the erythema/eschar scores at 24, 48 and 72 hours was 3.3, 2.0 and 3.7 in animal 1, 2 and 3, respectively. The mean of the edema scores at 24, 48 and 72 hours was 3.7, 2.0 and 4.0. The skin was normal within 8 days after patch removal in 2/3 animals respectively within 10 days after the end of exposure in the third animal. In conclusion, the substance has to be regarded as irritating to the skin.

 

Eye irritation

The primary eye irritant effect of the registration item was investigated in a GLP study according to OECD TG 405 (1995). Three female albino rabbits were exposed to 0.1 mL of the test article in the left eye. The eyes were examined and the changes were graded according to a numerical scale 1, 24, 48, 72 hours and on day 7 after dosing. After the treatment, only slight changes were observed in the eyes. The following mean values, based on the results from the 24, 48 and 72 hour readings, were calculated: Cornea opacity: 0/4, iris lesion: 0/2, redness of conjunctiva: 1/3, edema of conjunctiva: 0.8/4. It is concluded that the test substance caused no eye irritation under the conditions of the test.

Justification for classification or non-classification

Skin irritation

The mean value for erythema/eschar and for edema from grading at 24, 48 and 72 hours after patch removal was ≥ 2,3 - ≤ 4,0 in 2 of the tested animals.

 

Based on the above stated assessment of the skin irritation potential, the substance needs to be classified as skin irritant Category 2 according to Regulation (EC) No. 1272/2008 (CLP).

 

Eye irritation

All effects were fully reversible within 7 days and the mean scores following grading at 24, 48 and 72 hours after installation of the test material were below 1 for corneal opacity and iritis and below 2 for conjunctival redness and conjunctival edema. According to Regulation (EC) No. 1272/2008 (CLP), the substance has thus not to be classified as irritating to the eyes.