N,N'-methylenediacrylamide

Endpoint summary

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

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

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

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

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

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).

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.

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

50 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented report of a guideline study conducted to GLP.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 433 draft (Acute Inhalation Toxicity: Fixed Concentration Procedure) (not officially approved)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed concentration procedure

Limit test:

yes

Species:

rat

Strain:

Fischer 344

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Raleigh, NC
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 205-217 g
- Fasting period before study: no data
- Housing: microisolator cages, 2 rats /cage
- Diet: NIH-07 dietand ad libitum
- Water: reverse osmosis water ad libitum
- Acclimation period: 15 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.8-26.1°C
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours:12 hours
IN-LIFE DATES: From: 16/05/2000 To: 16/05/2000

Route of administration:

inhalation

Type of inhalation exposure:

nose only

Vehicle:

other: nitrogen

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 52-position Cannon nose-only exposure system (Lab Products, Maywood, NJ)
- Exposure chamber volume:
- Method of holding animals in test chamber:
- Source and rate of air:
- Method of conditioning air:
- Treatment of exhaust air: activated charcoal
- Temperature, humidity, pressure in air chamber:
TEST ATMOSPHERE
- Brief description of analytical method used: A GC was equipped with a splitless injection port and two test columns, connected together (for better resolution) and a flame ionization detector were used to monitor the nose-only exposure system. The GC was not calibrated because no reliable method was found to make GC standards; however, earlier studies indicated that GC was a quick, useful method to monitor acrylamide concentration stability and 0.5 mL samples of acrylamide vapour were analyzed at approximate 15-minute intervals to monitor system stability. An HPLC was used to determine the vapour concentration of acrylamide. Vapour samples of 25 mL were taken at approximate 30-minute intervals, dissolved in 150 µL of water and then analyzed by HPLC. The samples taken by this method were the primary concentration measurements because the analysis was completed closer to real-time than the impinger method.
- Samples taken from breathing zone: yes
VEHICLE
- Composition of vehicle (if applicable): nitrogen with humified oxygen
- Justification of choice of vehicle: inert
- Purity: >99%

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

6 h

Concentrations:

5.6 ppm (16 mg/m3)

No. of animals per sex per dose:

10 males

Control animals:

no

Details on study design:

- Duration of observation period following administration: 24 hours
- Frequency of observations and weighing: Observations: 15 minutes, 6 hours and 24 hours post administration. Weighing at beginning of study
- Necropsy of survivors performed: yes
- Other examinations performed: gross necropsy

Statistics:

Not applicable as there were no mortalities.

Sex:

male

Dose descriptor:

LC0

Effect level:

> 5.6 ppm

Based on:

test mat.

Exp. duration:

6 h

Mortality:

None

Clinical signs:

other: None

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In a study to investigate the acute toxicity of acrylamide to rats following exposure by the inhalation route, there were no mortalities or systemic effects at 5.6 ppm (16 mg/m3) which was the highest air concentration attainable. It can be concluded that the LC0 for acrylamide in rats by inhalation is greater than 5.6 ppm (16 mg/m3).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

12.1 mg/m³ air

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1975

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documeted study (pre-GLP). Complete study report with analytical data.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

no

Remarks:

Pre-GLP

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

2 male and 2 female rabbitts
24 hours prior to application the entire trunk of the test animals was clipped

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

Unabraided skin

Duration of exposure:

24 hours

Doses:

200, 795, 1580 and 3160 mg/kg of 50.7% aqueous acrylamide solution

No. of animals per sex per dose:

2 male and 2 female rabbits/group

Control animals:

no

Details on study design:

The animals were observed for mortality and clinical signs for 14 days.

Statistics:

None given

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 141 mg/kg bw

95% CL:

> 710 - < 1 851

Remarks on result:

other: 2250 mg test material/kg bw

Mortality:

200 mg test material/kg = 0/4
795 mg test material/kg = 0/4
1580 mg test material/kg = 1/4
3160 mg test material/kg = 3/4

Clinical signs:

other: At the two highest exposure levels tremors and incoordination of hindlimbs were noted, and, in addition, the surviving female at 1,612 mg/kg was in poor condition and lost weight. All other animals showed some bodyweight gain at the end of the observation

Gross pathology:

No microscopic observations were conducted

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute dermal LD50 for acrylamide is 1141 mg acrylamide/kg bw

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 141 mg/kg bw