Reaction mass of N,N-ethanediylbis- Hexadecanamide, N-[2-[(1-oxohexadecyl)amino]ethyl]-Octadecanamide, Hexadecanoic acid, 12-hydroxystearic acid, reaction products with ethylenediamine, N,N-ethanediylbis-Octadecanamide and Octadecanoic acid, 12-hydroxystearic acid, reaction products with ethylenediamine

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of N,N-ethanediylbis- Hexadecanamide, N-​[2-​[(1-​oxohexadecyl)​amino]​ethyl]​-Octadecanamide, Hexadecanoic acid, 12-​hydroxystearic acid, reaction products with ethylenediamine, N,N-ethanediylbis-Octadecanamide and Octadecanoic acid, 12-​hydroxystearic acid, reaction products with ethylenediamine

EC Number:

906-763-4

Molecular formula:

not applicable, mixture of substances

IUPAC Name:

Reaction mass of N,N-ethanediylbis- Hexadecanamide, N-​[2-​[(1-​oxohexadecyl)​amino]​ethyl]​-Octadecanamide, Hexadecanoic acid, 12-​hydroxystearic acid, reaction products with ethylenediamine, N,N-ethanediylbis-Octadecanamide and Octadecanoic acid, 12-​hydroxystearic acid, reaction products with ethylenediamine

Test material form:

other: Nearly white solid

Test animals

Species:

rat

Strain:

other: HanRcc: WIST (SPF)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services CH-4414 Füllinsdorf
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 12 weeks
- Fasting period before study: fasted for approximately 17 hours
- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse mainte-nance diet, batch no. 001/06 (Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad libitum.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum
- Acclimation period: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
From: 30.5. To: 22.06.2006

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Number of animals per group: 3 females
Total number of animals: 6 females

Control animals:

yes

Details on study design:

DOSE FORMULATION:
The test item was pulverised into a fine powder at RCC Ltd Itingen with a centrifugal mill 1000 (retsch ZM 1000; Serie-Nr.: 30379024). The test item was mixed with liquid nitrogen approximately 10 minutes before milling and was added bit by bit to the centrifugal mill. Thereafter, the test item was dried during 12 hours in an exsiccator at room temperature. The dose formulations were made shortly before each dosing occasion using a magnetic stirrer and a spatula as homogenizers.
The test item reduced into a fine powder was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight: volume). Homogeneity of the test item in the vehicle was maintained during administrantion using a magnetic stirrer.

TREATMENT:
The animals received a single dose of the test item by oral gavage administration at 2000 mg/kg body weight after being fasted for approximately 17 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing. The application volume was 10 mL/kg body weight.
Rationale: Oral administration was considered to be an appropriate application method as it is a possible route of human exposure during manufacture, handling and use of the test item.

OBSERVATIONS:
- Mortability/Viability: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.
- Body weights: On test days 1 (prior to administration), 8 and 15.
- Clinical signs: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15. All abnormalities were recorded.

PATHOLOGY:
- Necropsy: All animals were killed at the end of the observation period by Carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained.
- Statistical analysis: No statistical analysis was used.
- Data compilation: Body weights were on-line. Clinical signs were recorded on data sheets. Mortability/viability were compiled into the RCC Tox Computer System during recording and/or recorded on data sheets. Macroscopic findings were compiled into the RCC Tox Computer System during recording. The RCC Tox Computer System (RCC-Tox-Lims) had been validated with respect to data collection, storage and retrievability.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other:

Remarks:

after single oral administration to female rats, observed over a period of 14 days.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: Slightly ruffled fur was noted in all animals of the first treated group from the 1-hour reading to test day 3. Hunched posture was also noted in the same animals from the 2- or 3-hour reading to the 5-hour reading nd slight sedation was also noted in two

Gross pathology:

No macroscopic findings were recorded at necropsy.

Applicant's summary and conclusion

Interpretation of results:

other: not classified

Conclusions:

The median lethal dose of the test substance after single oral administration to female rats, observed over a period of 14 days is LD50 (female rat) > 2000 mg/kg body weight.

Executive summary:

Two groups, each of three female HanRcc:WIST (SPF) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (PEG 300) at a concentration of 0.2 g/mL and administered at a volume dosage of 10 mL/kg.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 -15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2 -15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period.

Slightly ruffled fur was noted in all animals of the first treated group from the 1 -hour reading to test day 3. Hunched posture was also noted in the same animals from the 2- or 3 -hour reading to the 5 -hour reading and slight sedation was also noted in two animals at the 3- and 5 -hour reading.

In the second treated group, slightly ruffled fur was noted in all animals from the 2- to the 5 -hour reading. Hunched posture was present in the same animals from the 1 -hour to the 3 -or 5 -hour reading. Slight sedation was noted in one animal at the 3 -hour reading and in the two remaining animals from the 2- to the 5 -hour reading.

The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.