Registration Dossier

Administrative data

Description of key information

Skin irritation:Weight-of-evidence approach using 6 in vivo skin irritation studies performed according to OECD guideline 404.

Eye irritation: Weight-of-evidence approach using 10 studies performed according to or similar to OECD guideline 405.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
From March 7, 2006 to April 11, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
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: Granja Cunícola San Bernardo, S.L. (Tulebras, Navarra, Spain)
- Age at study initiation: 10-13 weeks old
- Weight at study initiation: 2.4-3.0 kg
- Housing: The animals were housed individually in stainless steel cages with a grille floor placed on racks.
- Diet (e.g. ad libitum): standard rabbit diet (SAFE 112C; Batches: 50602 and 51114), ad libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-25 ºC
- Humidity (%): 40-70%
- Photoperiod (hrs dark / hrs light): 12 hours of light nad 12 hours of darkness every 24 hours.


Type of coverage:
semiocclusive
Preparation of test site:
shaved
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:
60 minutes, 24, 48, 72 hours, 7 days and 14 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 6 cm2
- Type of wrap if used: adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the remains of the test item were removed with distilled water.
- Time after start of exposure: 4 hours
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
2.11
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0.78
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: 24-72 h
Score:
2.67
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: 24-72 h
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 24-72 h
Score:
1.67
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 24-72 h
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 24-72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 24-72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
No data
Other effects:
A loss of 30 grams in body weight was recorded between the day of administration and the 24 hours following it in the first rabbit administered, as was a loss of 110 grams in body weight between 48 and 72 hours after the administration. A loss of 70 grams in body weight was recorded in another animal between 72 hours and 7 days after the administration, and a loss of 10 grams between the day of administration and the 24 hours afterwards in the case of the remaining animal. However, a posterior recovery of the weight was observed in all the animals.

The behaviour and physical state of the rabbits was normal throughout the study.
Interpretation of results:
GHS criteria not met
Conclusions:
The means of the erythema and edema evaluation 24, 48 and 72 hours after removing the patch in the animals after a 4-hour exposure to the test item, were 2.11 (erythema) and 0.78 (edema). Only one animal showed a mean score (24-72 h) greater than 2.3. Based on the CLP classification criteria, the test material should not be classified for skin irritation. Based on the DSD classification criteria, the test material should be classified as skin irritant, R38.
Executive summary:

The degree of primary dermal irritation caused by the test item was determined after application to the shaven skin of 3 rabbits. The study was carried out according to OECD guideline 404 and EU method B.4. Initially one animal was used, and 3 semiocclusive patches were applied sequentially during 3 minutes, 1 and 4 hours, respectively. A confirmation assay was then sequentially performed, applying a patch with the test item during 4 hours in another two animals. The means of the erythema and edema evaluations at 24, 48 and 72 hours after removing the patch in the animals after a 4 -hour exposure to the test item were 2.11 (erythema) and 0.78 (edema).

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
From October 26, 1999 to November 16, 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
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: David Percival Ltd, Moston, Sandbach, Cheshire, UK
- Age at study initiation: 12 to 16 weeks old
- Weight at study initiation: 2.48 to 3.09 kg
- Housing: The animals were individually housed in suspended metal cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: a minimum of five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 ºC
- Humidity (%): 30 to 70 %
- Air changes (per hr): the rate was approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours of darkness.
Type of coverage:
semiocclusive
Preparation of test site:
other: clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL of the test material.

Duration of treatment / exposure:
4 hour
Observation period:
14 days
Number of animals:
3 animals
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: the trunk of the rabbit was wrapped in an elasticated corset (TUBIGRIP)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Any residual test material was removed by gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirits.
- Time after start of exposure: 4 hours after application (when the patch was removed).

SCORING SYSTEM:
according to the OECD guideline nº 404.
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: 24-72 h
Score:
0
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: 24-72 h
Score:
0
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 24-72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 24-72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Loss of skin elasticity and loss of skin flexibility. At day 7, crust formation was observed. At day 14, slight desquamation was observed.
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 24-72 h
Score:
1.67
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 24-72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Loss of skin elasticity and loss of skin flexibility. At day 7, crust formation was observed. At day 14, slight desquamation was observed.
Irritant / corrosive response data:
No corrosive effects were noted.
Other effects:
No data

Individual skin reactions following 4-hours exposure period

 

Skin Reaction

Observation

Time

Individual Scores -Rabbit Number and Sex

(Bodyweight kg)

Total

Erythema/Eschar Formation

 

1Male

(3.09)

198 Male

(2.74)

200 Female

(2.48)

 

1 Hour

0

2

2

( 4 )

24 Hours

0

2

2

4

48 Hours

0

2Le

2le

( 4 )

72 Hours

0

2LeLf

2LeLf

4

7 Days

-

OCf

OCf

( 0 )

1 4 Days

-

OD

OD

( 0 )

Oedema Formation

1Hour

0

2

2

( 4 )

24 Hours

0

1

2

3

48 Hours

0

1

2

( 3 )

72Hours

0

1

1

2

7 Days

-

0

0

( 0 )

14 Days

-

0

0

( 0 )

Sum of 24 and 72-hour Readings (S)                 13

Primary Irritation Index                               13/6 - 2.2

( ): total values not used for calculation of the primary index

Cf: crust formation

D: slight desquamation

Le: loss of skin elasticity

Lf: loss of skin flexibility

- : observation not required

 

Individual daily and mean scores for dermal irritation following 4-hour exposure

 

 

Skin Reaction

Reading (Hours)

Individual Scores - Rabbit Number and Sex

(Bodyweight kg)

 

1Male

(3.09)

198Male

(2.48)

200Female

(2.74)

 

Erythema/Eschar Formation

24

0

2

2

48

0

2

2

72

0

2

2

total

 

0

6

6

Mean Score

 

0.0

2.2

2.0

Oedema Formation

24

0

1

2

48

0

1

2

72

0

1

1

Total

 

0

3

5

Mean Score

 

0.0

1.0

1.7

 

Interpretation of results:
GHS criteria not met
Conclusions:
Well-defined erythema was noted at two treated skin sites one hour after patch removal and at the 24, 48 and 72-hour observations. Slight oedema was noted at two treated skin sites one hour after patch removal. Very slight oedema was noted at two treated skin sites at the 72-hour observation. One treated skin site appeared normal throughout the study. Crust formation was noted at two treated skin sites at the 7-day observation with slight desquamation at these two treated skin sites at the 14-day observation. No corrosive effects were noted.
According to the CLP classification criteria, the test material should not be classified as skin irritant. According to the DSD classification criteria, the test material should be classified as skin irritant, R38.
Executive summary:

The Acute Dermal Irritation/corrosion assay for the test substance was performed in New Zealand rabbits. 0.5 mL of the test material was applied to a site of about 2.5 x 2.5 cm of intact skin of 3 rabbits and covered by semi-occlusive dressing. The administration of the test substance was performed in a sequential manner. The first animal was exposed to the test substance stepwise at three sites of the skin: site 1 for three minutes, site 2 for 1 hour and site 3 for 4 hours. Any residual test substance was removed by swabbing with cotton wool soaked in 74% Industrial Methylated Spirits. After considerations of the skin reactions produced on the first animal, the remaining two animals were exposed to the test substance for 4 hours. The treated areas of the animals were examined for erythema/eschar and oedema as well as other local skin alterations approximately 1, 24, 48, and 72 hours after patch removal. Additional observations were made on days 7 and 14 to assess the reversibility of skin reactions. Dermal alterations were scored according to OECD guideline nº 404. Well-defined erythema was noted at two treated skin sites one hour after patch removal and at the 24, 48 and 72-hour observations. Slight oedema was noted at two treated skin sites one hour after patch removal. Very slight oedema was noted at two treated skin sites at the 72-hour observation.One treated skin site appeared normal throughout the study.Crust formation was noted at two treated skin sites at the 7-day observation with slight desquamation at these two treated skin sites at the 14-day observation. No corrosive effects were noted.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
from September 9, 1980 to September 19, 1980
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Well-documented non-GLP study performed similar to OECD TG 404. Exposure lasted for 24h (instead of 4h) and the observation period was limited to 7 days (instead of 14 days).
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
24h exposure; observation period limited to 7 days; type of wrap unclear
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kuiper's Rabbitry, Gary, IN, USA
- Age at study initiation: 14 weeks
- Weight at study initiation: 2.4 - 3.0 kg
- Housing: individually in screen bottom cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
no data
Type of coverage:
other: gauze patch overwrapped with saran wrap and elastoplast tape
Preparation of test site:
other: shaved and abraded
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:
24 hours
Observation period:
7 days
Number of animals:
3 males, 3 females
Details on study design:
TEST SITE
- Area of exposure: no data
- % coverage: no data
- Type of wrap if used: each treated area was covered with a 5x5 cm gauze patch and overwrapped with saran wrap and elastoplast tape to maintain the test material in contact with the skin and decrease of evaporation

REMOVAL OF TEST SUBSTANCE
- Washing (if done): wiping (not washing)
- Time after start of exposure: 24h

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal: 1-6
Time point:
other: 24h, 72h, 96h after start of exposure
Score:
1 - 4
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal: 1-6
Time point:
other: 24h, 72h, 96h after start of exposure
Score:
0 - 4
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Irritant / corrosive response data:
Blanching of the test sites was observed in 3 animals at the 24h reading. Subcutaneous hemorrhage was observed within the test sites of 2 animals at the 72h, and 96h readings, and in 1 animal at the 7d reading. Fissuring of the test sites was observed by 1 animal at 72h, 2 animals at 96h and 3 animals at 7d. Eschar formation was observed in 1 animal at 7d.
Other effects:
No data
Interpretation of results:
study cannot be used for classification
Conclusions:
After 24h of exposure to the test material, the treated skin sites showed severe erythema and severe edema in a few animals. No conclusion can be drawn on the classification according to CLP and DSD as the observation period is limited to 7 days and consequently no conclusion can be drawn on reversibility of effects.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
From 10 April 1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Well-documented GLP study performed similar to OECD TG 404. An occlusive patch was used instead of a semi-occlusive patch. No 14 day observation period.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
Occlusive patch was used; 7 day observation period
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: A. Smith, Warlingham, Surrey, UK
- Age at study initiation: no data
- Weight at study initiation: 2.6 - 3.1 kg
- Housing: individually housed in grid bottomed metal cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22
- Humidity (%): 53-65
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:
occlusive
Preparation of test site:
shaved
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:
7 days
Number of animals:
6 females
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- % coverage: ca. 6 cm2
- Type of wrap if used: test material was plasced evenly over a lint pad, which was placed onto the skin and held in position by a strip of 'Sleek' plastic adhesive strapping.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): gently cleaned using cotton wool soaked in water at 37°C
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (grade 2)
Irritation parameter:
edema score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.28
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
At the 72h observation, desquamation from the skin surface was observed in 3 animals.
At the 7d observation, 5 animals showed desquamation from the skin surface, of which one also showed well-defined erythema.
Other effects:
No data
Interpretation of results:
study cannot be used for classification
Conclusions:
No corrosive effects have been observed after 4h occlusive exposure to the test substance. However, as the observation period was limited to 7 days, no conclusion can be drawn on the reversibility of effects and consequently no conclusion can be drawn on the classification according to CLP or DSD.
Endpoint:
skin corrosion: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not indicated
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Well-documented non-GLP study performed similar to OECD TG 405. No 21-day observation period.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No 21d observation period
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 2.4-2.6 kg
- Housing: individual cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18°C
- Humidity (%): 50-60%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: other eye served as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: no data
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 6 days
Remarks on result:
other: 1 animal showed grade 1 on day 4 and 5
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.39
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.33
Max. score:
3
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: 1 animal showed grade 1 on days 4-7
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.39
Max. score:
4
Reversibility:
fully reversible within: 2 days
Irritant / corrosive response data:
In 5 of 6 animals all effects were reversible within 5 days. In 1 animal iris effects appeared on day 4 and 5 (grade 1) and chemosis appeared on days 4-7, which was not reversible.
Other effects:
No data
Interpretation of results:
study cannot be used for classification
Conclusions:
Reversible skin effects were observed in 5 of the 6 animals. In 1 animal chemosis was observed which was not reversible. As there was no 21-day observation period, no conclusion can be drawn on the classification according to CLP and DSD.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not indicated
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Well-documented non-GLP study performed similar to OECD TG 405. No 21-day observation period.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No 21d observation period
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 2.4-2.6 kg
- Housing: no data
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18°C
- Humidity (%): 50-60%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: other eye served as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: no data
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.72
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (score 2)
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.28
Max. score:
2
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (score 1)
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.89
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (grade 1)
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.56
Max. score:
3
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (grade 2)
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.22
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (grade 1)
Irritant / corrosive response data:
In 5 of 6 animals all effects were reversible within 4 days. In 1 animal significant effects were observed which were not reversible after 7 days.
Other effects:
No data
Interpretation of results:
study cannot be used for classification
Conclusions:
Reversible skin effects were observed in 5 of the 6 animals. In 1 animal more severe effects were observed which were not reversible. As there was no 21-day observation period, no conclusion can be drawn on the classification according to CLP and DSD.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
from September 9, 1980 to September 30, 2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Well-documented non-GLP study performed similar to OECD TG 405.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Hoppers Unlimited, Verona, Wisconsin, USA
- Age at study initiation: ca. 14 weeks
- Weight at study initiation: 2.9-3.5 kg
- Housing: individually in screen bottom cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
no data
Vehicle:
unchanged (no vehicle)
Controls:
other: other eye served as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
6 animals
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein (3, 7, 14, 21 days)
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.31
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2.56
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2.17
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2.11
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritant / corrosive response data:
Blanching of the conjunctivae was exhibited in all animals at 48h and in 1 animal at 72h. One animal was observed with small hemorrhage of the conjunctivae at 48h. Pannus was observed in five animals at the 7 day observation. Corneal neovascularization was exhibited by 2 animals at 7 days, 4 animals at 14 days and in 5 animals at 21 days.
Other effects:
One animal was found dead on day 12. Necropsy showed no lesions but bloody nasal discharge. The cause of death could not be determined.
Interpretation of results:
Category 1 (irreversible effects on the eye)
Conclusions:
As irreversible eye effects were noted in the 5 remaining animals after 21 days, the substance should be classified for irreversible eye effects, as category 1 serious eye damage (according to CLP Regulation).
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
9-30 September 1980
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Well-documented non-GLP study performed similar to OECD TG 405.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Hoppers Unlimited, Verona, Wisconsin, USA
- Age at study initiation: ca. 14 weeks
- Weight at study initiation: 2.6-3.3 kg
- Housing: individually in screen bottom cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
no data
Vehicle:
unchanged (no vehicle)
Controls:
other: other eye served as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
6 animals
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein (3, 7, 14, 21 days)
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.42
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.67
Max. score:
2
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2.58
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2.06
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.67
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritant / corrosive response data:
Pannus was observed at 3 animals at 7 days. Two animals were observed with corneal neovascularization at the 14 and 21 day observations.
Other effects:
No data
Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
As irreversible eye effects were noted in 4 animals after 21 days, the substance is considered to have irreversible eye effects and should be classified as serious eye damage cat 1 according to CLP regulation. .
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Not indicated
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP study performed according to OECD TG 405. No 21-day observation period.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No 21d observation period
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harald Schriever, Kaninchenfarm, 2740 Bremervörde, Neuendamm 88
- Age at study initiation: no data
- Weight at study initiation: 2.4-2.8 kg
- Housing: in batteries (40x45x50 cm)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 2
- Humidity (%): 50-85%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 120 lux; 12/12

Vehicle:
water
Controls:
other: other eye served as control
Amount / concentration applied:
The test substance was diluted in water (14.3 g in 100 g) to get a 5% solution. The pH was 5.5.
Duration of treatment / exposure:
Single treatment
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein at 1 and 7 days.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0.5
Max. score:
2
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 3 animals (grade 1)
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.56
Max. score:
3
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 3 animals (grade 1 or 2)
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.5
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 3 animals (grade 1 or 2)
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.33
Max. score:
3
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: not fully reversible in 1 animal (grade 1)
Irritant / corrosive response data:
No data
Other effects:
No data
Interpretation of results:
study cannot be used for classification
Conclusions:
Reversible skin effects were observed in 3 animals (reversible after 3 days). In 3 animals effects on the iris and conjunctivae were observed which were not reversible after 7 days. As there was no 21-day observation period, no conclusion can be drawn on the classification according to CLP and DSD.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
From 21 November 1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Well-documented GLP study performed similar to OECD TG 405. Additional animals were tested to assess the effect of rinsing. Observations of the eye already ended after 72h.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
an additional group of 3 animals was tested at which eyes were rinsed after 5 sec after instillation. No 21d observation period.
Qualifier:
according to
Guideline:
other: U.S. Federal Hazardous Substances Act (U.S. Federal Register, 1973, vol. 38, no. 187, section 1500:32 and revised annually in the US Code of Federal Regulations, Title 16 Part 1500, Subch C)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: recognized breeders whose standards are acceptable to the Animal Welfare Department of Toxicol Laboratories.
- Age at study initiation: no data
- Weight at study initiation: 1.9 - 2.2 kg
- Housing: grid bottomed galvanised metal cages
- Diet (e.g. ad libitum): 90-95 g pelleted diet
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: other eye as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
72h
Number of animals or in vitro replicates:
9
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): the eyes of 3 animals were washed 5 sec after instillation; the eyes of 6 animals were not washed.

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: standard light source
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 (unrinsed) animals
Time point:
other: 24, 48, 72h
Score:
0.06
Max. score:
4
Reversibility:
fully reversible within: 48h
Remarks on result:
other: Dulling of cornea of 1 rabbit at 24h
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 (unrinsed) animals
Time point:
other: 24, 48, 72h
Score:
0.28
Max. score:
2
Reversibility:
fully reversible within: 72h
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 (unrinsed) animals
Time point:
other: 24, 48, 72h
Score:
0.94
Max. score:
3
Reversibility:
not fully reversible within: 72h
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 (unrinsed) animals
Time point:
other: 24, 48, 72h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48h
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 (unrinsed) animals
Time point:
other: 24, 48, 72h
Score:
0.83
Max. score:
4
Reversibility:
not fully reversible within: 72h
Irritant / corrosive response data:
The remaining effects at 72h were only grade 1 (chemosis in 2 animals; redness in 1 animal).
Other effects:
No data
Interpretation of results:
GHS criteria not met
Conclusions:
Given the remaining effects after 72h, it is expected that full reversibility would occur after 21 days. According to the classification criteria of both CLP and DSD, the substance does not need to be classified.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not indicated
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Well-documented non-GLP study performed similar to OECD TG 405.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No 21 day observation period
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: ca. 14 weeks
- Weight at study initiation: 2.5 - 3.5 kg
- Housing: individually in screen bottom cages in air conditioned quarters
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
-no data
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye served as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
14 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residue of test material and accumulate discharge were flushed from the eyes each time they were scored.

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: no data
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2.72
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
No data
Other effects:
No data
Interpretation of results:
irritating
Conclusions:
According to the classification criteria of CLP and DSD, the substance should be classified as an eye irritant (CLP Cat. 2 and R36).
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not indicated
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Non-GLP study performed similar to OECD TG 405. No 21 day observation period.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No 21 day observation period
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: ca. 14 weeks
- Weight at study initiation: 2.5 - 3.5 kg
- Housing: individually in screen bottom cages in air conditioned quarters
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
-no data
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye served as control
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
single treatment
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residue of test material and accumulate discharge were flushed from the eyes each time they were scored, however no special washing was done.

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: no data
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72h
Score:
1.17
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
mean
Remarks:
6 animals
Time point:
other: 24, 48, 72 h
Score:
0.72
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
No data
Other effects:
No data
Interpretation of results:
Category 2A (irritating to eyes)
Conclusions:
According to the classification criteria of CLP, the substance should be classified as an eye irritant (category 2). According to the classification criteria of DSD, the substance should not be classified for eye irritation.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
14 February 2006 - 4 April 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Granja Cunícula San Bernardo, S.L
- Age at study initiation: 13-14 weeks
- Weight at study initiation: 2.8 kg
- Housing: in a stainless steel cage with a grille floor and placed on a rack
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 40-65%
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
The treated eye was washed with physiological saline solution at one hour after administration
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
1
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
other: neovascularization and growth of the surface tissue over cornea
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
hyperaemia
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
2.67
Max. score:
3
Reversibility:
not reversible
Remarks on result:
other: bloody aspect of conjunctiva and nictitating membrane at 1h
Irritation parameter:
conjunctivae score
Remarks:
edema
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
4
Max. score:
4
Reversibility:
not reversible
Irritant / corrosive response data:
After 1 h: scattered or diffuse areas of corneal opacity (grade 1); conjunctiva and the nictitating membrane presented a bloody aspect, conjunctiva showed diffuse beefy colouring (grade 3) and swelling with lids more than half closed (grade 4); lacrimation with moistening of the lids and adjacent fur with bloody aspect (grade 2); iris could not be examined.
After 24, 48, 72h: similar effects as after 1 h.
After 7 days: easily discernible translucent area in the cornea (25-50% of the area) and neovascularization; diffuse beefy colouring in the conjunctiva, swelling with the lids half closed; lacrimation with moistening of the lids and adjacent fur.
After 14 and 21 days: nacreous area of opacity in the cornea (75-100% of the area) and neovascularization; diffuse crimson colouring in the conjunctiva, swelling with the lids half closed; lacrimation with moistening of the lids and adjacent fur and whitish secretion.
After 21 days: growth of the surface tissue over the cornea.
Other effects:
No data
Interpretation of results:
Category 1 (irreversible effects on the eye)
Conclusions:
The test substance is considered to be severe irritant to the eye according to DSD regulation, and serious eye damage 1 according to CLP regulation.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
From November 8, 1999 to November 22, 1999
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
OECD guideline 405. GLP study. Although the test substance did not produce severe effects in the single animal tested, two additional animals were not tested.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
only 1 animal tested, while no severe effects were observed and consequently two additional animals could have been tested
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
Purity: approximately 79%
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: David Percival Ltd, Moston, Sandbach, Cheshire, UK
- Age at study initiation: 12 to 16 weeks old
- Weight at study initiation: 2.95 Kg
- Housing: The animal was housed in suspended metal cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: a minimum of five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 ºC
- Humidity (%): 30 to 70 %
- Air changes (per hr): the rate was approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours of darkness.
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of the test substance
Duration of treatment / exposure:
72 hours.
Observation period (in vivo):
72 hours. Additional observations were made on days 7 and 14 to assess the reversibility of the ocular effects.
Number of animals or in vitro replicates:
One animal.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing was done

SCORING SYSTEM: The ocular reactions were scored by Draize Scale.
1. CONJUNCTIVAE
(A) Redness (refers to palpebral and bulbar conjunctivae excluding cornea and ins)
Vessels normal: 0
Vessels definitely injected above normal: 1
More diffuse, deeper crimson red, individual vessels not easily discernible: 2
Diffuse beefy red: 3

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

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

THE TOTAL SCORE = (A + B + C) x 2 MAXIMUM TOTAL = 20

2. IRIS
(D) Values
Normal: 0
Folds above normal, congestion, swelling, circumcorneal injection: 1
(any or all of these or combination of any thereof) ins still reacting to light
(sluggish reaction is positive)
No reaction to light, haemorrhage, gross destruction (any or all of these): 2

THE TOTAL SCORE = D x 5 MAXIMUM TOTAL = 10

3. CORNEA
(E) Degree Of Opacity (most dense area used)
No opacity: 0
Scattered or diffuse areas, details of ins dearly visible: 1
Easily discernible translucent areas, details of iris slightly obscured: 2
Opalescent areas, no details of ins visible, size of pupil barely discernible: 3
Opaque, ins invisible: 4

(F)Area of Cornea Involved
One quarter (or less) but not zero: 1
Greater than one quarter but less than half: 2
Greater than half but less than three quarters: 3
Greater than three quarters, up to whole area: 4

THE TOTAL SCORE = (E x F) x 5 MAXIMUM TOTAL = 80

MAXIMUM TOTAL SCORE POSSIBLE = 110


TOOL USED TO ASSESS SCORE: a standard ophthalmoscope
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 24-48-72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 24-48-72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 24-48-72 h
Score:
2.3
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 24-48-72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
The treated eye appeared normal at the 14 day observation.
Other effects:
No data

Table 1: Individual scores and total scores for ocular irritation 

Rabbit Number and Sex

IPR - 3

(Bodyweight kg)

10 Male (2.95)

Time After Treatment

1

hour

24 hours

48 hours

72

hours

7

days

14

days

CORNEA

 

 

 

 

 

 

Degree of Opacity

0d

1

2

3

V

0

Area of Opacity

2

4

3

3

1

0

IRIS

1

1

1

1

0

0

CONJUNCTIVAE

 

 

 

 

 

 

A - Redness

2

2

2

3

2

0

B - Chemosis

2

2

2

2

1

0

C - Discharge

3

3

3

3

2

0

 d: dulling of the normal lustre of the cornea

 V: vascularisation, localized in growth of vessels for approximately 1 to 2 mm

Interpretation of results:
study cannot be used for classification
Conclusions:
The test substance produces ocular irritation in one rabbits The treated eye appeared normal at the 14-day observation.
As only 1 animal has been tested, no definite conclusion can be drawn on the classification according to CLP and DSD.
Executive summary:

The Acute Eye Irritation/Corrosion study for the test substance was performed in New Zealand White rabbits. 0.1 mL of the test material was instilled into the conjunctival sac of the right eye. The eye was held closed for about one second to prevent loss of the test substance. The left eye remained untreated and served as a control. Assesment of ocular damage/irritation was made immediately after the administration of the test material, and approximately after 1 hour, 24, 48 and 72 hours following treatment. Additional observations were made on days 7 and 14 to assess the reversibility of the ocular effects. Based on the results, the test substance was considered to be severe irritant so the test was performed only in one rabbit. One hour after treatment a dulling of the normal lustre of the cornea, iridial inflammation, and conjunctival moderate irritation were noted. Irial inflammation was persisted for 72 hours. Severe conjuctivial irritation and opalescent opacity in the cornea were noted at 72 hours after treatment. Vascularisation, with a localised in growth of vessels for approximately 1 to 2 mm, was noted at the 7-day observation. The treated eye appeared normal at the 14-day observation. The test substance produces ocular irritation in rabbits. The treated eye appeared normal at the 14-day observation.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

The Garcia (2006) and Sanders (2000) studies are GLP studies according to OECD guideline 404 and EU guideline B.4.

Garcia tested a 30% pure solution while Sanders tested a 79% pure solution. These two studies had an observation period of 14 days. The other reliable studies did not observe skin effects up to 14 days after exposure. However, the results of these studies as well as of the non reliable studies were consistent.

Based on the available data, the test materials would be classified as irritating according to Directive 67/548/EEC. However, taking into account the criteria established under CLP Regulation, the substance would not be classified.

Since no data is available on the pure substance, taking into account the worst case, the proposal is to classify the substance as a Skin Irritant Category 2.

Eye irritation:

There are ten studies available performed according to or similar to OECD guideline 405.

The only Klimisch 1 scored study (Garcia, 2006) was performed using a 30% solution of the test substance in water. Only 1 animal was tested, which showed irreversible effects even after 21 days of observation. According to this study the substance should be classified as Eye Dam 1 H318 (CLP) and R41 (DSD).

Sanders (2000) performed a study using a 79% pure substance on 1 animal only. As the observed effects were reversible and not severe enough to allow classification as corrosive to the eyes, the substance was tested in 2 additional animals. No definitive conclusion can be drawn on the classification based on this study.

Several solutions have been tested in the 7 remaining studies. The 3 studies by Sterner et al. did not observe the animals for 21 days, so no conclusions can be drawn on reversibility of effects and therefore no definite classification can be derived. In the three studies solutions of 5%, 10% and 20% were tested, having pH values of 5.5, 8.0 and 4.9, respectively, which seems not logical.

Two studies by Thompson using solutions of 5% and 35% showed irreversible eye effects after 21 days. Based on these studies the substance should be classified as Eye Dam 1 H318 (CLP) and R41 (DSD).

An eye irritation study by Haynes (1983) showed reversible eye effects, although the observation period was only 72 hours. Given the slight effects it can be expected that the effects are fully reversible at 21 days. Therefore, the test substance (probably a 30% solution) should not be classified for eye irritation.

Two studies by the Goldschmidt GmbH (1976) showed full reversibility of the effects within 7 days (5% solution tested) or 14 days (30% solution tested). According to the criteria of CLP, both solutions should be classified as an eye irritant.

Taking all eye irritation data together, the results are sometimes conflicting. Mostly dilutions between 5% and 35% have been tested, which sometimes showed irreversible eye effects. Therefore, it is concluded that the pure substance should be classified as Eye Dam 1 H318 (CLP) and R41 (DSD).

Justification for classification or non-classification

Skin irritation: Based on the available data, the substance is classified as irritating according to Directive 67/548/EEC. However, taking into account the criteria established under CLP Regulation, the substance would not be classified.

Since the tested substance had a purity/concentration of 79% and since no data is available on the pure substance, taking into account the worst case, the proposal is to classify the substance as a Skin Irritant Category 2 H315.

Eye irritation: Based on the above weight-of-evidence approach, the substance is classified as severe eye irritant (R41) according to Directive 67/548/EEC and as Eye Dam. 1 H318 according to CLP.