Registration Dossier

Administrative data

Description of key information

Skin irritation: No skin irritation was caused by 4 hours exposure to EA-3098. EA-3098 does not have to be classified as a skin irritant.

Eye irritation: Instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours. EA-3098 does not have to be classified as an eye irritant.

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:
August 23rd 2005 to September 2nd 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviation from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.
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
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF Guidelines (2000) including the most recent partial revisions
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: At least 6 weeks old
- Weight at study initiation: At least 1 kg
- Housing: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56 x 44 x 37.5 cm)
- Diet: Approximately 100 g per day of standard laboratory diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany).
- Water: ad libitum access to tap water
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C +/- 3°C (actual range: 20.2 - 22.3°C)
- Humidity (%): 30 - 70% (actual range: 40 - 77%)
- Air changes (per hr): Approximately 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

Type of coverage:
semiocclusive
Preparation of test site:
other: clipped free of fur
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
Each animal was treated by dermal application of 0.5 grams of the test substance. The test substance was moistened with watery ethanol (50% v/v) immediatley before application, to ensure close contact with the animals skin.

Duration of treatment / exposure:
4 h
Observation period:
72 hours
Number of animals:
3 males
Details on study design:
STUDY DESIGN:
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 7 days later, after considering the degree of skin irritation observed in the first animal.

TREATMENT:
Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10 x 15 cm2). Whenever considered necessary the treated skin areas were re-clipped at least 3 hours before the observations, to facilitate scoring.

Each animal was treated by dermal application of 0.5 grams of the test substance. The test substance was moistened with 1 ml watery ethanol (50% v/v) and applied to the skin of one flank, using a metalline patch of 2 x 3 cm. The patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage.

Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water.

After the final observation, the animals were sacrificed by intra-venous injection of pentobarbital.

OBSERVATIONS:
Mortality/Viability: Twice daily
Toxicity: At least once daily
Body weight: Day of treatment (prior to application) and at termination.
Irritation: The skin reactions were assessed at appromixmately 1, 24, 48 and 72 hours after the removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded. Adjacent areas of the untreated skin of each animal served as controls.













Irritation parameter:
erythema score
Basis:
mean
Remarks:
( Animal 754)
Time point:
other: Mean of scores at 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(Animal 744)
Time point:
other: Mean of scores at 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(Animal 751)
Time point:
other: Mean of scores at 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
edema score
Basis:
mean
Remarks:
(Animal 754)
Time point:
other: Mean of scores at 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
edema score
Basis:
mean
Remarks:
(Animal 744)
Time point:
other: Mean of scores at 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
edema score
Basis:
mean
Remarks:
(Animal 751)
Time point:
other: Mean of scores at 24, 48 and 72 hrs
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritant / corrosive response data:
No skin irritation was caused by 4 hours exposure to EA-3098.
Other effects:
No staining of the treated skin by the test substance was observed and no test substance remnants were seen.

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.
Interpretation of results:
not classified
Conclusions:
No skin irritation was caused by 4 hours exposure to EA-3098. Based on the results of the study EA-3098 does not have to be classified for skin irritation.
Executive summary:

A skin irritation/corrosion study with EA-3098 in the rabbit (4 -hour semi-occlusive application) was conducted according to the following guidelines:

- OECD 404 "Acute Dermal Irritation/Corrosion" (2002)

- EC, Council Directive 67/548/EEC, Annex V, B.4 (2004) "Acute Toxicity: Dermal Irritation/Corrosion"

- US EPA OPPTS 870.2500 (1998) Acute Dermal Irritation

- JMAFF Guidelines (2000) including the most recent partial revisions

Three rabbits were exposed to 0.5 grams of EA-3098, applied onto clipped skin for 4 hours using a semi-occlusive dressing. Skin reactions were assessed 1, 24, 48 and 72 hours after exposure.

No skin irritation was caused by 4 hours exposure to EA-3098.

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Based on these results, EA-3098 does not have to be classified as a skin irritant as it does not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP).

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:
30th Agust 2005 to 8th September 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviation from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.
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
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF guidelines (Japan)
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: Charles river Deutschland, Kisslegg, Germany
- Age at study initiation: At least 6 weeks old
- Weight at study initiation: At least 1 kg
- Housing: housed individually in labeled cages with perforated floors (Scanbur, Denmark, dimensions 56 x 44 x 27.5 cm).
- Diet: Approximately 100 g per day of standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany).
- Water: ad libitum access to tap water
- Acclimation period: at least five days prior to start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 +/- 3.0°C (actual range: 20.2 - 22.3°C)
- Humidity (%): 30 - 70% (actual range 40 - 77%)
- Air changes (per hr): approximately 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light followed by 12 hours darkness per day

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
Animals were treated by instillation of, on average 49.8 mg (range 49.4 - 50.1 mg) of the test substance (a volume of approximately 0.1 ml), in the conjunctival sac of one of the eyes.

Duration of treatment / exposure:
Single application for 72 hour exposure.
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
3 males.
Details on study design:
pH MEASUREMENT:
pH determined as 8.4.

STUDY DESIGN:
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner one week later, after considering the degree of eye irritation observed in the first animal.

TREATMENT:
Animals were treated by instillation of, on average, 49.8 mg (range 49.4 - 50.1 mg) of the test substance (a volume of approximately 0.1 ml), in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

Immediately after the 24-hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

After the final observation, the animals were sacrificed by intra-venous injection of pentobarbital.

OBSERVATIONS:
Mortality/Viability: Twice daily
Toxicity: At least once daily
Body weight: Day of treatment (prior to installation) and at termination.
Irritation: The eyes of each animal were examned approximately 1, 24, 48 and 72 hours after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.

The irritation was assessed according to the numercial Draize Scale for Scoring Occular Irritation.








Irritation parameter:
cornea opacity score
Remarks:
(Degree of opacity)
Basis:
mean
Remarks:
Animal 752
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
cornea opacity score
Remarks:
(Degree of opacity)
Basis:
mean
Remarks:
( Animal 741)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
cornea opacity score
Remarks:
(Degree of opacity)
Basis:
animal: (Animal 757)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
iris score
Basis:
mean
Remarks:
(Animal 752)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
2
Reversibility:
other: no effects observed
Irritation parameter:
iris score
Basis:
mean
Remarks:
(Animal 741)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
2
Reversibility:
other: no effects observed
Irritation parameter:
iris score
Basis:
mean
Remarks:
(Animal 757)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
2
Reversibility:
other: no effectss observed
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
(Animal 752)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
other: Score at 1 hr = 2, score at 24 hr = 2, score at 48 hr = 1, score at 72 hr = 0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
(Animal 741)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
other: Score at 1 hr = 2, score at 24 hr = 2, score at 48 hr = 1, score at 72 hr = 0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
(Animal 757)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
other: Score at 1 hr = 2, score at 24 hr = 1, score at 48 hr = 1, score at 72 hr = 0
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
(Animal 752)
Time point:
other: Mean of scores at 24, 48 & 72 hours
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 24 hrs
Remarks on result:
other: Score of 1 at 1 hr
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
(Animal 741)
Time point:
other: Mean of scores at 24, 58 & 72 hours
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 hrs
Remarks on result:
other: Score at 1 hr = 1, score at 24 hr = 1, score at 48 hr = 0
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
(Animal 757)
Time point:
other: Mean of scores at 24, 48 & 72 hrs
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 24 hrs
Remarks on result:
other: Score of 1 at 1 hr
Irritant / corrosive response data:
Irritation (Table 1 - attached background material):
Instillation of approximately 50 mg of EA-3098 (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours.

No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% flurescein, 24 hours after the test substance instillation revealed no corneal epithelial damage.

There was no evidence of ocular corrosion.
Other effects:
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Remnants of the test substance were present in the eye on Day 1.
Interpretation of results:
not classified
Conclusions:
Instillation of approximately 50 mg of EA-3098 (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours.
No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% flurescein, 24 hours after the test substance instillation revealed no corneal epithelial damage.
EA-3098 does not have to be classified for eye irritation.
Executive summary:

An acute eye irritation/corrosion study was conducted on the material, EA-3098, according to the following guidelines:

OECD No. 405 (2002) "Acute Eye Irritation/Corrosion"

EC, Council Directive 67/548/EEC, Annex V, B5 (2004) "Acute Toxicity: Eye irritation / corrosion"

EPA, OPPTS 870.2400 (1998) "Acute Eye Irritation"

JMAFF guidelines (2000); including the most recent partial revisions.

Single samples of approximately 50 mg of EA-3098 (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation.

Instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours.

Overall results calculated on the basis of the scores for each animal at 24, 48 and 72 hours were:

 

Mean score 1

Mean score 2

Mean score 3

Max. value observed

Max. duration of any effect

Max. value at the end of the observation period

Redness

1.0

1.0

0.7

2

48 hours

0

Chemosis

0.0

0.3

0.0

1

24 hours

0

Cornea

0.0

0.0

0.0

0

-

0

Iris

0.0

0.0

0.0

0

-

0

Remnants of the test substance were present in the eye on Day 1.

Based on these results EA-3098 does not have to be classified as an eye irritant as it does not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP)

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation

Introduction:

A skin irritation/corrosion study with EA-3098 in the rabbit (4 -hour semi-occlusive application) was conducted according to the following guidelines:

- OECD 404 "Acute Dermal Irritation/Corrosion" (2002)

- EC, Council Directive 67/548/EEC, Annex V, B.4 (2004) "Acute Toxicity: Dermal Irritation/Corrosion"

- US EPA OPPTS 870.2500 (1998) Acute Dermal Irritation

- JMAFF Guidelines (2000) including the most recent partial revisions

Method:

Three rabbits were exposed to 0.5 grams of EA-3098, applied onto clipped skin for 4 hours using a semi-occlusive dressing. Skin reactions were assessed 1, 24, 48 and 72 hours after exposure.

Results:

No skin irritation was caused by 4 hours exposure to EA-3098.

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Conclusion:

Based on these results, EA-3098 does not have to be classified as a skin irritant as it does not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP).

Eye Irritation

Introduction:

An acute eye irritation/corrosion study was conducted on the material, EA-3098, according to the following guidelines:

OECD No. 405 (2002) "Acute Eye Irritation/Corrosion"

EC, Council Directive 67/548/EEC, Annex V, B5 (2004) "Acute Toxicity: Eye irritation / corrosion"

EPA, OPPTS 870.2400 (1998) "Acute Eye Irritation"

JMAFF guidelines (2000); including the most recent partial revisions.

Method:

Single samples of approximately 50 mg of EA-3098 (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation.

Results:

Instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours.

Overall results calculated on the basis of the scores for each animal at 24, 48 and 72 hours were:

 

Mean score 1

Mean score 2

Mean score 3

Max. value observed

Max. duration of any effect

Max. value at the end of the observation period

Redness

1.0

1.0

0.7

2

48 hours

0

Chemosis

0.0

0.3

0.0

1

24 hours

0

Cornea

0.0

0.0

0.0

0

-

0

Iris

0.0

0.0

0.0

0

-

0

Remnants of the test substance were present in the eye on Day 1.

Conclusion:

Based on these results EA-3098 does not have to be classified as an eye irritant as it does not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP)

Justification for selection of skin irritation / corrosion endpoint:

The study has been conducted according to OECD Guideline 404 and GLP and is adequately reported. The study has been assigned a reliability 1.

Justification for selection of eye irritation endpoint:

The study has been conducted according to OECD Guideline 405 and GLP and is adequately reported. The study has been assigned a reliability 1.

Justification for classification or non-classification

The results from skin irritation and eye irritation studies were evaluated according to the Classification, Labelling and Packaging Regulation (CLP).

Skin irritation:

No skin irritation was caused by 4 hours exposure to EA-3098. Based on these results, EA-3098 does not have to be classified as a skin irritant as it does not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP).

Eye irritation:

Instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours.

The overall results calculated on the basis of the scores for each animals at 24, 48 and 72 hours did not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP).