N,N'-methylenediacrylamide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Start of the experimental phase July 19, 2016; Termination of the in-life phase August 17, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

EU Method B.1 tris (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 (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-methylenediacrylamide

EC Number:

203-750-9

EC Name:

N,N'-methylenediacrylamide

Cas Number:

110-26-9

Molecular formula:

C7H10N2O2

IUPAC Name:

N,N'-methylenediacrylamide

Test material form:

solid: crystalline

Details on test material:

Purity = 99.8%

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7,97633 Sulzfeld, Germany
- Age at study initiation: Approx. 8 - 9 weeks
- Weight at study initiation: 164 - 200 g
- Fasting period before study: Feeding was discontinued approx. 16 hours before administration; only tap water was then available ad libitum.
- Housing: During the 14-day observation period the animals were kept in groups of 3 animals in MAKROLON cages (type III plus)
- Diet (e.g. ad libitum): Feeding was discontinued approx. 16 hours before administration; only tap water was then available ad libitum.
- Water (e.g. ad libitum): tap water ad libitum.
- Acclimation period: At least 5 adaptation days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C (maximum range)
- Humidity (%): relative humidity of 55% ± 15% (maximum range)
- Air changes (per hr): 15 to 20 times
- Photoperiod (hrs dark / hrs light): The rooms were lit (about 150 lux at approx. 1.50 m room height) and darkened for periods of 12 hours each.

IN-LIFE DATES: From:First dosing July 25, 2016 To: Termination of the in-life phase August 17, 2016

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.8% aqueous hydroxypropylmethylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.8%
- Amount of vehicle (if gavage):
- Justification for choice of vehicle: 0.8% aqueous hydroxypropylmethylcellulose was chosen as vehicle as it is known not to produce toxic effects.
- Lot/batch no. (if required): Methocel; batch no. 13D 03-N03, Fagron GmbH & Co., 22885 Barsbüttel, Germany
- Purity:

MAXIMUM DOSE VOLUME APPLIED: The administration volume was 20 mL/kg b.w.

DOSAGE PREPARATION (if unusual):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The dose used is selected from a series of defined dose levels.

Doses:

2000 mg N,N‘-Methylenediacrylamide/kg b.w.
300 mg N,N‘-Methylenediacrylamide/kg b.w.
50 mg N,N‘-Methylenediacrylamide/kg b.w.

No. of animals per sex per dose:

15 female animals
3 dose level group of 3 (2000 mg/kg b.w.) or 6 female animals (300 and 50 mg/kg b.w.)
Administration volume 20 mL/kg b.w.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 1 test day and 2 recovery weeks
- Frequency of observations and weighing: Observations were performed before and immediately, 5, 15, 30 and 60 min, as well as 3, 6 and 24 hours after administration. Individual body weights were recorded before administration of the test item and thereafter in weekly intervals up to the end of the study and at death.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, macroscopical / necropsy findings
other: During the follow-up period of two weeks, changes of skin and fur, eyes and mucous membranes, respiratory and the circulatory, autonomic and central nervous system and somatomotor activity as well as behaviour pattern were observed at least once a day until all symptoms subsided, thereafter each working day. Attention was also paid to possible tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Observations on prematurely deceased animals were made at least once daily to minimize loss of animals during the study. The time of death was recorded as precisely as possible. Individual body weights were recorded before administration of the test item and thereafter in weekly intervals up to the end of the study and at death. Changes in weight were calculated and recorded.
At the end of the experiments, all animals were sacrificed, dissected and inspected macroscopically. All gross pathological changes were recorded.

Statistics:

No statistical analysis could be performed (the method used is not intended to allow a calculation of a precise LD50 value).

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

approximate LD50

Effect level:

>= 50 - <= 300 mg/kg bw

Based on:

test mat.

Mortality:

2000 mg N,N‘-Methylenediacrylamide/kg b.w. All 3 animals died prematurely.
300 mg N,N‘-Methylenediacrylamide/kg b.w. Four of the six animals died prematurely.
50 mg N,N‘-Methylenediacrylamide/kg b.w. No animal died prematurely

Clinical signs:

other: 2000 mg N,N‘-Methylenediacrylamide/kg b.w. Reduced motility, ataxia, dyspnoea, salivation, pilo-erection, cyanosis, red liquid in the eyes in all 3 animals. 300 mg N,N‘-Methylenediacrylamide/kg b.w. Reduced motility, ataxia, tremor, increased muscle tone,

Gross pathology:

No macroscopical findings were noted at autopsy in any dose level.

Applicant's summary and conclusion

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

Under the present test condition, the test item achieved an LD50 value between 50 and 300 mg N,N‘-Methylenediacrylamide/kg b.w.