1-Piperazineacetamide, N-(2,6-dimethylphenyl)-

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 October 2014 - 20 November 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method according to OECD Guideline 420. GLP study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-(2,6-dimethylphenyl)-2-(piperazin-1-yl)acetamide

EC Number:

610-916-8

Cas Number:

5294-61-1

Molecular formula:

C14H21N3O

IUPAC Name:

N-(2,6-dimethylphenyl)-2-(piperazin-1-yl)acetamide

Details on test material:

- Name of test material (as cited in study report): N-(2,6-dimethylphenyl)-1-piperazineacetamide)
- Physical state: white solid
- Lot/batch No.: M13856C
- Analytical purity: 99.8%
- Expiration date of the lot/batch: December 2015

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 10-week-old
- Weight at study initiation: 197.5 g (average body weight)
- Fasting period before study: about 18 hours before the administration of the test item but restored 3 hours after the administration of the test item.
- Housing: plastic cages covered with wire bar lids (dimensions: 58 x 37 x 21 cm), with UV-sterilized wood shavings as bedding.
- Diet (e.g. ad libitum): “Murigran” standard granulated laboratory food, ad libitum.
- Water (e.g. ad libitum): tap water, ad libitum.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 – 23°C
- Humidity (%): 40 – 75%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 60 mg/mL
- Amount of vehicle (if gavage): 1 mL

MAXIMUM DOSE VOLUME APPLIED: 0.5 mL/100 g bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The starting dose for the sighting study was selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg bw as a dose expected to produce evident toxicity. Since no data were available, the sighting study commenced with the administration of the test item at a dose of 300 mg/kg bw to one female rat. Since no evident toxicity was produced, the test item at a single dose of 2000 mg/kg bw was administered to the second animal. On the grounds of the sighting study results (the animal given 2000 mg/kg bw died on the first day of the observation period), the dose of 300 mg/kg bw was selected to be used in the main study.

Doses:

Sighting study: 300 and 2000 mg/kg bw
Main study: 300 mg/kg bw

No. of animals per sex per dose:

2 females in the sighting study.
4 females in the main study.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: The evaluation of general condition of the animals, i.e. the observation of all animals for morbidity and mortality was conducted twice a day or once a day (on days off) during the 14-day experiment. Body weights of the animals were determined on days 0 (directly before the administration of the test item), 7, and 14 (before euthanasia)
- Necropsy of survivors performed: yes.
- Other examinations performed:
Detailed clinical observations: on the day of the test item administration (day 0), i.e. 10, 30, and 60 minutes after the administration and then at hourly intervals up to the 5th hour after the administration. From the 1st to the 14th day of the experiment, the detailed clinical observations were performed once a day.
Gross examinations: After the 14-day observation period, all animals were euthanized by intraperitoneal administration of morbital at a dose of 200 mg/kg bw and subjected to gross examinations. The detailed gross examinations involved the observation of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with their contents.

Results and discussion

Preliminary study:

Clinical signs: No signs of toxicity were observed following single administration of the test item at a dose of 300 mg/kg bw. Following single administration of the test item at a dose of 2000 mg/kg bw signs of toxicity were found on day 0. The signs, i.e. a slight decrease in locomotor activity and tremors, occurred 3 hours post-administration. The animal died on the first day of the observation period.
Body weights: Body weight of the surviving animal (dosed 300 mg/kg bw) increased.
Gross examinations: The gross examinations of the accidently dead and the euthanized females did not reveal any lesions.

Effect levels

Sex:

female

Dose descriptor:

discriminating dose

Effect level:

300 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived the experiment.

Clinical signs:

other: No signs of toxicity were found.

Gross pathology:

No pathological changes were observed.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Migrated information

Conclusions:

The substance was classified as category 4 according to CLP Regulation (EC) no. 1272/2008 since the dose of 2000 mg/kg bw caused death in the sighting study. No effects were observed at 300 mg/kg bw (main study).

Executive summary:

The acute oral toxicity study based on a fixed dose method was performed according to OECD Guideline 420 (GLP study). The experiment commenced with a sighting study in which the test item was first administered to a female rat at a single dose of 300 mg/kg bw but, since no signs of toxicity were observed, a dose of 2000 mg/kg bw was given to a second rat. A slight decrease in locomotor activity and tremors occurred 3 hours post-administration. The animal died on the first day of the observation period. On the grounds of these results, four animals in the main study were given the test item at a dose of 300 mg/kg bw. After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment. Body weights of the animals were also determined. After the 14-day observation period, the animals were euthanized and subjected to a necropsy and a detailed gross examination. No signs of toxicity were found and all animals survived the experiment. During the 14-day experiment, body weights of the animals increased. Regarding the gross examinations, no lesions were found in the animals. The substance was classified as category 4 according to CLP Regulation (EC) no. 1272/2008 since the dose of 2000 mg/kg bw caused death in the sighting study.