N,N''-(methylenedi-4,1-phenylene)bis[N'-octyl]urea

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 03 - 26, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

451-060-3

EC Name:

-

Cas Number:

122886-55-9

Molecular formula:

Hill formula: C31H48N4O2 CAS formula: C31H48N4O2

IUPAC Name:

N,N’’-(methylenedi-4,1-phenylene)bis(N’-octylurea)

Constituent 2

Reference substance name:

N-{4-[(4-{[(octadecylamino)carbonyl]amino}phenyl)methyl]phenyl}-N’-octylurea

Cas Number:

117328-86-6

Molecular formula:

Hill formula: C41H68N4O2 CAS formula: C41H68N4O2

IUPAC Name:

N-{4-[(4-{[(octadecylamino)carbonyl]amino}phenyl)methyl]phenyl}-N’-octylurea

Constituent 3

Reference substance name:

3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea

EC Number:

406-690-3

EC Name:

3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea

Cas Number:

43136-14-7

Molecular formula:

Hill formula: C51H88N4O2 CAS formula: C51H88N4O2

IUPAC Name:

N,N’’-(methylenedi-4,1-phenylene)bis(N’-octadecylurea)

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): KY-EU
- Appearance: white powder
- Storage condition of test material: in darkness at room temperature

Test animals

Species:

rat

Strain:

other: Sprague-Dawley Rj: SD (IOPS Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approx. 6 weeks old
- Weight at study initiation: mean body weight 181 ± 6 g for the males and 175 ± 6 g for the females
- Housing: The animals were group housed in polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm). Each cage contained one to seven animals of the same sex during the acclimation period and three rats of the same sex and group during the treatment period. Each cage contained autoclaved sawdust (SICSA, Alfortville, France).
- Diet (e.g. ad libitum): Free access to A04 C pelleted diet (UAR, Villemoisson, Epinay-sur-Orge, France)
- Water (e.g. ad libitum): Free access to drinking water filtered by a FG Millipore membrane (0.22 micron)
- Acclimation period: The acclimatization period was at least 5 days before the start of treatment

Sawdust is analysed by the supplier for composition and contaminant levels. Each batch of food is analysed by the supplier for composition and contaminant levels. Bacteriological and chemical analyses of water are performed regularly by external laboratories. These analyses include the detection of possible contaminants (pesticides, heavy metals and nitrosamines).

No contaminants were known to have been present in the diet, drinking water or bedding material at levels which may be expected to have interfered with or prejudiced the outcome of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 30 - 70%
- Air changes (per hr): approximately 12 (filtered, non-recycled air)
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 03 September 2002 to 26 September 2002

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% methylcellulose

Details on oral exposure:

GAVAGE METHOD: metal gavage tube fitted to a 5 ml glass syringe

VEHICLE
- Justification for choice of vehicle: No data

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight.

DOSAGE PREPARATION: The test item was prepared at the chosen concentration in the vehicle. Each test item preparation was made freshly on the morning of administration by the CIT Pharmacy and any unused material was discarded that same day.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: As no information on the toxic potential of the test item was available, for animal welfare reasons, the starting dose-level of 200 mg/kg bw was chosen. Three males were used in the initial step.
After the first assay, as no mortality occurred, another assay was carried out on three males at the dose-level 2000 mg/kg bw.
After the second assay, as no mortality was observed, the results were confirmed in three females at the dose-level 2000 mg/kg bw.

Doses:

Initial step 200 mg/kg body weight.
First and second assay 2000 mg/kg body weight.

No. of animals per sex per dose:

200 mg/kg bw: 3 males
2000 mg/kg bw: 3 males and 3 females

Control animals:

no

Details on study design:

The animals were fasted for an overnight period of approximately 18 hours before dosing. Water was available ad libitum. Food was given back approximately 4 hours after administration of the test item.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Time of death was recorded individually, in terms of the number of hours or days after dosing.
Body weights: The animals were weighed individually just before administration of the test item on day 1 and then on days 8 and 15.
Clinical signs: The animals were observed frequently during the hours following administration of the test item for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day.
- Necropsy of survivors performed: On day 15, all animals were killed by carbon dioxide asphyxiation. All study animals were subjected to a macroscopic examination as soon as possible after death. After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed.
- 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

Preliminary study:

No deaths occurred in the initial step of 200 mg/kg body weight.

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No clinical signs were observed in any of the treated animals.

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Other findings:

None.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

KY-EU was administered by oral gavage to two subsequent groups of three male Sprague-Dawley rats at 200 and 2000 mg/kg body weight and three female rats at 2000 mg/kg body weight. The study was conducted according to OECD 423 and GLP guidelines.

The oral LD50 value of KY-EU in Sprague-Dawley rats was established to exceed 2000 mg/kg body weight.

Based on these results, KY-EU does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.