Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to guideline
Guideline:
other: Commission Regulation (EC) No 440/2008
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Qualifier:
according to guideline
Guideline:
other: Japan MAFF Testing Guideline of 12 Nosan No. 8147
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N''-hexane-1,6-diylbis[N'-benzylurea]
EC Number:
254-274-3
EC Name:
N,N''-hexane-1,6-diylbis[N'-benzylurea]
Cas Number:
39072-70-3
Molecular formula:
C22H30N4O2
IUPAC Name:
3-benzyl-1-{6-[(benzylcarbamoyl)amino]hexyl}urea
Details on test material:
- Name of test item: N,N''-Hexane-1,6-diylbis[N'-benzylurea]
- Test item No.: 11/0726-1
- Batch identification: 259-635
- Purity: 98.0 g/100 g determined by 1H NMR spectroscopy using the internal standard method
- Homogeneity: The test item was homogeneous by visual inspection.
- Storage stability: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
- Expiry date: January 09, 2013
- Storage conditions: Room temperature; no direct sunlight; protect against humidity
- Physical state / color: Solid / white

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: approx. 10 weeks
- Weight at study initiation: Administration group 1: 163 - 178 g; administration group 2: 180 - 184 g
- Fasting period before study: at least 16 hours before administration, but water was available ad libitum.
- Housing: Single housing in Makrolon cage, type III with bedding (H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)) and enrichment (NGM E-022; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria)
- Diet (e.g. ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): approx. 10
- Day / night rhythm: 12 h / 12 h

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Olive oil Ph.Eur
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20 g/100 mL
- Justification for choice of vehicle: Good homogeneity in olive oil Ph.Eur.
- Amount of vehicle (if gavage): 10 mL/kg bw

DOSAGE PREPARATION
The test item was ground with mortar and pestle. The test item preparation was produced shortly before application by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer. Additionally the homogeneity of the test item preparation during application was ensured by stirring with a magnetic stirrer.
Form of application: Suspension

SELECTION OF DOSES/CONCENTRATIONS
By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. Because no mortality occurred, 2000 mg/kg bw were administered to another group of 3 female animals in the second step.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
3 female animals per administration group
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation;recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Mortality at least once each workday.
- Necropsy of survivors performed: yes
- Other examinations performed: No histological examinations were performed.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred in both test groups.
Clinical signs:
No clinical signs were observed in both test groups during clinical examination.
Body weight:
The mean body weight of the test groups increased throughout the study period within the normal range.
Gross pathology:
There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period.

Applicant's summary and conclusion