Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Description: Clear slightly yellow very viscous liquid
- Purity: 100%
- Storage condition of test material: In refrigerator (2-8°C) in the dark

Test animals

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

Administration / exposure

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).

Results and discussion

Effect levels
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.

Applicant's summary and conclusion

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.