Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: The oral LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

Acute dermal toxicity: The dermal LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

The test item does not meet the criteria for classification according to the Classification, Labelling and Packaging Regulation (CLP).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16th August 2005 to 1st 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.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF guidelines (2000) including the most recent partial revisions
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Silzfeld, Germany.
- Age at study initiation: Approximately 9 weeks
- Weight at study initiation: 169 - 205 g. Body weight did not exceed +/- 20% of the sex mean.
- Fasting period before study: Food was withheld overnight (for a maximum of 20 hours) prior to dosing until 3-4 hours after administration of the test substance.
- Housing: 3 animals per cage in labelled Macrolon cages (MIV type: height 18 cm), containing sterilised sawdust as bedding material and paper as cage enrichment
- Diet: ad libitum access to standard pelleted laboratory animal diet (from Altromin (code VRF 1), Lage, Germany).
- Water: ad libitum access to tap water
- Acclimation period: at least five days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0°C +/-3.0°C (actual range: 20.5 - 22.5°C
- Humidity (%): actal range: 41-85%
- Air changes (per hr): approximately 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN-LIFE DATES: From: Day 1 - day of dosing To: Day 15 - terminal sacrifice
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
VEHICLE
Propylene glycol was used as the vehicle and was selected based on trial formulations performed and on test substance data supplied by the sponsor.


MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

DOSAGE PREPARATION (if unusual): The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle.



Doses:
2000 mg/kg
No. of animals per sex per dose:
2 groups of 3 females, total of 6 (at 2000 mg/kg)
Control animals:
no
Details on study design:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females.
The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: Observations were made for mortality/viability twice daily. Bodyweights were recorded on Day 1 (pre-administration), Day 8 and Day 15.

- Necropsy of survivors performed: yes - at the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

- Other examinations performed:
Clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded.
Statistics:
No statistical analysis was performed.
Preliminary study:
Not applicable
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture, uncoordinated movements and piloerection
were noted in the animals on days 1 and 2.
Body weight:
The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
Effects on organs:
No abnormalities were found at macroscopic post mortem
examination of the animals.
Other findings:
None

The oral LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline on the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg dody weight.
According to the OECD 423 test guideline the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.
Executive summary:

Assessment of acute oral toxicity with EA-3098 in the rat (Acute Toxic Class Method) was carried out based on the guidelines described in:

-OECD 423 (2001) "Acute Toxicity-Oral, Acute Toxic Class Method",

-EC Council Directive 67/548/EEC, Annex V, B.1 tris (2004) "Acute Oral Toxicity",

-EPA OPPTS 870.1100 (2002), "Acute Oral Toxicity - Acute Toxic Class Method"

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

EA-3098 was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred.

Hunched posture, uncoordinated movements and piloerection were noted in animals on Days 1 and 2.

The body weight gain shown by the animals over the study period was considered to be normal.

No abnormalities were found at macroscopic ppost mortem examination of the animals.

The oral LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Based on these results EA-3098 does not have to be classified and has no obligatory labelling requirement for oral toxicity according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
The study has been conducted according to OECD Guideline 423 and GLP and is adequately reported. The study has been assigned a reliability 1.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24th August 2005 to 7th 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.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
Protocol deviation: No body weight determined of the male found dead on Day 2. the study data not adversely affected by this deviation.
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF Guidelines (2000), including the most recent revisions
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles river Deutschland, Sulzfeld, Germany.
- Age at study initiation: Approximately 8 weeks
- Weight at study initiation: Males: 255 - 281 g, Females: 183-193 g. Body weight variation did not exceed +/-20% of the sex mean.
- Fasting period before study: None.
- Housing: Individually housed in labeled Macrolon cages (MIII type, height 18 cm), containing sterilized sawdust as bedding material and paper as cage-enrichment.
- Diet: Ad libitum access to standrd pelleted laboratory animal diet (from Altromin (code VRF 1), Lage, Germany).
- Water: ad libitum access to tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): actual range of 20.5 - 22.5°C
- Humidity (%): actual range of 41 - 85%
- 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
Vehicle:
propylene glycol
Details on dermal exposure:
TEST SUBSTANCE PREPARATION
Vehicle: Propylene glycol
Rationale: The vehicle was selected based on trial formulations performed and on test substance data supplied by the data.
Preparation: The formulation (w/w/) was prepared within 4 hours prior to dosing. Homogeneity was accopmplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle.


TEST SITE
- Clipping: One day before exposure an anrea of approximately 5 x 7 cm on the back of the animal was clipped.
- Area of exposure: back, 25 cmsquared for males and 18 cm quared for females
- % coverage: The formulation was applied in an area of approx. 10% of the total body surface
- Type of wrap if used: The formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (surgy 1D), succesively covered with aluminium foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Sfter 24 hours dressings were removed and the skin cleaned of residual test substance removed using tap water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Dose volume - 10 ml/kg bodyweight
- Concentration (if solution): Dose level - 2000 mg/kg body weight
- Constant volume or concentration used: yes

Duration of exposure:
Single dose with 24 hour contact period.
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males and 5 females at 2000 mg/kg.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: Observations were made for mortality/viability twice daily. Bodyweights were recorded on Day 1 (pre-administration), Day 8 and Day 15.

- Necropsy of survivors performed: yes - at the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

- Other examinations performed:
Clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded.
Statistics:
No statistical analysis performed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 1
One male was found dead on day 2 of treatment.
Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Clinical signs:
Flat or hunched posture, ptosis and/or chromodacryorrhoea
were noted in all males and some females on day 1 and/or 2.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No test substance related abnormalities were found at
macroscopic post mortem examination of the animals.

In one male diaphragmatic hernia of the liver was observed, which is an incidental finding occasionally noted among rats of this age and strain. This was therefore considered not toxicologically significant.
Other findings:
Scales and/or scabs were seen in the treated skin-area of
all females and one male during the observation period.

The dermal LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight
Executive summary:

Assessment of the acute dermal toxicity of EA-3098 in the rat was determined according to the following methods:

-OECD No. 402 (1987) "Acute Dermal Toxicity"

-EC Council directive 67/548/EEC, Annex V, B.3 (1992) "Acute Toxicity (Dermal)2

-EPA OPPTS. 870.1200 (1998), "Acute Dermal Toxicity"

-JMAFF Guidelines (2000), including the most recent revisions.

EA-3098 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

One male was found dead on Day 2 of treatment.

Flat or hunched posture, ptosis and/or chromodacryorrhoea were noted in all males and some females on Day 1 and/or Day 2.

Scales and/or scabs were seen in the treated skin-area of all females and one male during the observation period.

The body weight gain of the surviving animals during the observation period was within the range expected for rats used in this type of study.

No test substance related abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
The study has been conducted according to OECD Guideline 402 and GLP and is adequately reported. The study has been assigned a reliability 1.

Additional information

Acute Oral Toxicity

Introduction:

Assessment of acute oral toxicity with EA-3098 in the rat (Acute Toxic Class Method) was carried out based on the guidelines described in:

-OECD 423 (2001) "Acute Toxicity-Oral, Acute Toxic Class Method",

-EC Council Directive 67/548/EEC, Annex V, B.1 tris (2004) "Acute Oral Toxicity",

-EPA OPPTS 870.1100 (2002), "Acute Oral Toxicity - Acute Toxic Class Method"

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

Method:

EA-3098 was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

Results:

No mortality occurred.

Hunched posture, uncoordinated movements and piloerection were noted in animals on Days 1 and 2.

The body weight gain shown by the animals over the study period was considered to be normal.

No abnormalities were found at macroscopic post mortem examination of the animals.

Conclusion:

The oral LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline the D50 cut-off value was considered to exceed 5000 mg/kg body weight.

Acute Dermal Toxicity

Introduction:

Assessment of the acute dermal toxicity of EA-3098 in the rat was determined according to the following methods:

-OECD No. 402 (1987) "Acute Dermal Toxicity"

-EC Council directive 67/548/EEC, Annex V, B.3 (1992) "Acute Toxicity (Dermal)2

-EPA OPPTS. 870.1200 (1998), "Acute Dermal Toxicity"

-JMAFF Guidelines (2000), including the most recent revisions.

Method:

EA-3098 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

Results:

One male was found dead on Day 2 of treatment.

Flat or hunched posture, ptosis and/or chromodacryorrhoea were noted in all males and some females on Day 1 and/or Day 2.

Scales and/or scabs were seen in the treated skin-area of all females and one male during the observation period.

The body weight gain of the surviving animals during the observation period was within the range expected for rats used in this type of study.

No test substance related abnormalities were found at macroscopic post mortem examination of the animals.

Conclusion:

The dermal LD50 value of EA-3098 in Wistar rats was established to exceed 2000 mg/kg body weight.

Justification for selection of acute toxicity – oral endpoint

The available acute oral toxicity study is assigned as reliability study 1 and is the only acute oral toxicity study available.

Justification for selection of acute toxicity – dermal endpoint

The available acute dermal toxicity study is assigned as reliability study 1 and is the only acute dermal toxicity study available.

Justification for classification or non-classification

The substance does not meet the criteria for classification, under the CLP regulations, for acute toxicity via the oral or dermal routes based on the results of an acute oral toxicity study and an acute dermal toxicity study, which both gave LD50 results of >2000 mg/kg bodyweight.