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Administrative data

Description of key information

Acute oral and dermal studies with m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline performed in accordance with current OECD/EC test guidelines and GLP principles, showed LD50 values of 300-2000 mg/kg bw for acute oral toxicity, and of > 2000 mg/kg for acute dermal toxicity in rats.

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:
October 11, 2012 - November 06, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
(2001)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
(2002)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, April 2011, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 11-12 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean.
- Fasting period before study: overnight prior to dosing and until 3-4 hours after administration
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight.

DOSAGE PREPARATION: 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 and test substance (specific gravity of 1.220 was used for calculation). No correction was made for purity of the test substance.

Doses:
2000 and 300 mg/kg body weight

No. of animals per sex per dose:
2000 mg/kg: 3
300 mg/kg: 6 (2 groups of three females in a stepwise manner)
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:
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15 and at death (if found dead after Day 1).
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
- Other examinations performed: none.
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: The LD50 cut-off value was considered to be 500 mg/kg body weight.
Mortality:
All animals at 2000 mg/kg (3/3) were found dead on Day 1 or 2. No mortality was observed at 300 mg/kg.
Clinical signs:
All three animals at 2000 mg/kg showed hunched posture and piloerection on Day 1. One animal at 2000 mg/kg showed lethargy and uncoordinated movements on that day. No clinical signs were noted at 300 mg/kg.
Body weight:
The body weight gain shown by the surviving 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:
No abnormalities were found at macroscopic post mortem examination of the animals.
Beginning of autolysis was noted in two animals at 2000 mg/kg which were found dead on Day 2.
Interpretation of results:
harmful
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute oral toxicity study with rats, performed according to OECD 423 test guideline and GLP principles, an LD50 value was established to be within the range of 300-2000 mg/kg body weight.
Executive summary:

The test substance was assessed for toxicity in an acute oral toxicity study with female Wistar rats, performed according to OECD 423 test guideline and GLP principles. Initially, m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline was administered to three rats at 2000 mg/kg body weight. In a stepwise procedure two additional groups of females were dosed at 300 mg/kg body weight.

All animals at 2000 mg/kg (3/3) were found dead on Day 1 or 2. No mortality was observed at 300 mg/kg. All three animals at 2000 mg/kg showed hunched posture and piloerection on Day 1. One animal at 2000 mg/kg showed lethargy and uncoordinated movements on that day. No clinical signs were noted at 300 mg/kg. The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

No abnormalities were found at macroscopic post mortem examination of the surviving animals. Beginning of autolysis was noted in two animals at 2000 mg/kg which were found dead on Day 2.

Based on the results, the oral LD50 value of m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline in Wistar rats was established to be within the range of 300-2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

According to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
300 mg/kg bw
Quality of whole database:
The study has klimisch code 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:
November 29, 2012 - December 13, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
(1987)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
(1998)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, April 2011; including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar strain, Crl:WI (Han)
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 11 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean.
- Housing: Individually housed in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Vehicle:
propylene glycol
Details on dermal exposure:
One day before exposure (Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.

The formulation was applied in an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch, successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

Frequency: Single dosage, on Day 1.

Washing: Following application, dressings were removed and the skin cleaned of residual test substance using tap water.
Duration of exposure:
24 hours.
Doses:
2000 mg/kg


No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

Dose volume: 10 mL/kg body weight

DOSAGE PREPARATION: The formulation (w/w) was prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle (1.036) and specific gravity of the test substance (1.220 was used). No correction was
made for purity of the test substance.

Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: 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: none.
Statistics:
None.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched and/or flat posture, chromodarcryorrhoea and piloerection, were noted in the majority of animals on Days 1 and/or 2. One male showed ptosis and one female showed tremor on Days 1 and/or 2.
Body weight:
Five males and three females showed body weight loss in the first week. In the second week these animals recovered and showed body weight gain. The two remaining females showed body weight gain in the first and second week.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
General, focal or maculate erythema, necrosis, scabs and scales were seen in the treated skin-area of the animals during the observation period.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute dermal toxicity study with rats, performed according to OECD 402 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined.
Executive summary:

The test substance was assessed for toxicity in an acute dermal toxicity study with female Wistar rats, performed according to OECD 402 test guideline and GLP principles.

No mortality occurred. Hunched and/or flat posture, chromodarcryorrhoea and piloerection, were noted in the majority of animals on Days 1 and/or 2. One male showed ptosis and one female showed tremor on Days 1 and/or 2. General, focal or maculate erythema, necrosis, scabs and scales were seen in the treated skin-area of the animals during the observation period.

Five males and three females showed body weight loss in the first week. In the second week these animals recovered and showed body weight gain. The two remaining females showed body weight gain in the first and second week.

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

Based on the results the dermal LD50 value of m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline in Wistar rats was

established to exceed 2000 mg/kg body weight.

The test substance, m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has klimisch code 1.

Additional information

Oral:

The test substance was assessed for toxicity in an acute oral toxicity study with female Wistar rats, performed according to OECD 423 test guideline and GLP principles. Initially, m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline was administered to three rats at 2000 mg/kg body weight. In a stepwise procedure two additional groups of females were dosed at 300 mg/kg body weight.

All animals at 2000 mg/kg (3/3) were found dead on Day 1 or 2. No mortality was observed at 300 mg/kg.

All three animals at 2000 mg/kg showed hunched posture and piloerection on Day 1. One animal at 2000 mg/kg showed lethargy and uncoordinated movements on that day. No clinical signs were noted at 300 mg/kg. The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

No abnormalities were found at macroscopic post mortem examination of the survivinganimals. Beginning of autolysis was noted in two animals at 2000 mg/kg which were found dead on Day 2.

Based on the results, the oral LD50 value of m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline in Wistar rats was established to be within the range of 300-2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

Dermal:

The test substance was assessed for toxicity in an acute dermal toxicity study with female Wistar rats, performed according to OECD 402 test guideline and GLP principles.

No mortality occurred. Hunched and/or flat posture, chromodarcryorrhoea and piloerection, were noted in the majority of animals on Days 1 and/or 2. One male showed ptosis and one female showed tremor on Days 1 and/or 2. General, focal or maculate erythema, necrosis, scabs and scales were seen in the treated skin-area of the animals during the observation period.

Five males and three females showed body weight loss in the first week. In the second week these animals recovered and showed body weight gain. The two remaining females showed body weight gain in the first and second week.

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

Based on the results the dermal LD50 value of m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline in Wistar rats was

established to exceed 2000 mg/kg body weight.


Justification for selection of acute toxicity – oral endpoint
One study available.

Justification for selection of acute toxicity – dermal endpoint
One study available.

Justification for classification or non-classification

According to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline should be classified for acute oral toxicity as Category 4 and should be labeled as H302: Harmful if swallowed.

m-(2,3-epoxypropoxy)-N,N-bis(2,3-epoxypropyl)aniline does not have to be classified for acute dermal toxicity to Regulation (EC) No 1272/2008.