2-Propenamide, N-(2,2-dimethoxyethyl)-

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well performed study according to OECD technical guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

92/69/EEC

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: 93/21/EEC

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate attached to the study report.

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-(2,2-dimethoxyethyl)prop-2-enamide

EC Number:

610-472-5

Cas Number:

49707-23-5

Molecular formula:

C7 H13 N1 O3

IUPAC Name:

N-(2,2-dimethoxyethyl)prop-2-enamide

Details on test material:

- Name of test material (as cited in study report): N-(2,2-Dimethoxyethyl)acrylamide
- Chemical name: N-Acryloylaminoacetaldehyddimetylacetal, NAAADA
- Batch number: Z 3165-04
- Description: yellowish liquid
- Water solubility: completely mixable at 25°C
- Density: 1.065 g/cm3 at room temperature (determined in the testing facility)
- Storage: the test article was stored in a closed vessel at =<10°C in a refrigerator in the dark
- The test article was made to the room temperature prior using

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Wiga GmbH, D-97320 Sulzfeld
- Age at study initiation (start of acclimatisation): 35 - 42 days
- Weight 2days after arriving: Males: 191.5g +/- 6.9g (3.6%), n = 15
Females: 163.1g +/- 4.0g (2.5%), n = 15
- Fasting period before study: over night before administration
- Hygiene status upon supply: SPF
- Diet (e.g. ad libitum): Altromin 1326, pelleted standard diet, ad libitum, Batch: 140998/1206
- Water (e.g. ad libitum): tap water, ad libitum (municipal supply), Makrolon bottles, changed daily
- Acclimation period: 8/14/19 days before administration according to the different dose groups
- Identification: Ear notches and cage labelling showing the animal number, study number, dose, sex, time of dosing and end of observation period
- Randomisation: Animals were assigned according to random numbers one day prior to administration

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 - 25°C
- Humidity (%): 30 - 60%
- Air changes (per hr): Air conditioned, no further details given
- Photoperiod (hrs dark / hrs light): 12hrs / 12hrs


IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

- single dose
- in each case after an overnight fasting
- by oral gavage using a metal catheter and disposable plastic syringes
- individual doses were adjusted via the administered volume of the original test article according to the body weight on the day of administration

Doses:

Males [mg/kg body weight]: 2000 (0.188mL per 100g b.w.), 1500 (0141mL per 100g b.w.), 1000 (0.094mL per 100g b.w.)
Females [mg/kg body weight]: 1300 (0.122mL per 100g b.w.), 1150 (0.108mL per 100g b.w.), 1000 (0.094mL per 100g b.w.)

No. of animals per sex per dose:

5 animals per sex and dose

Control animals:

no

Details on study design:

See "Any other information on materials and methods incl. tables"

Statistics:

Body weights: Calculation of group mean values and standard deviations.
LD50 values: Probit analysis (Weber, E.: Grundriß der bilogischen Statistik, G. Fischer Verlag, Jena 1972)

Results and discussion

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50

Effect level:

ca. 1 199 mg/kg bw

Remarks on result:

other: See remarks on reults

Effect levels 2

Sex:

male

Dose descriptor:

approximate LD50

Effect level:

ca. 1 600 mg/kg bw

Remarks on result:

other: See remarks on reults

Mortality:

See tables 1 and 2
Comment on LD50, see "Remarks ond results including table and figures"

Clinical signs:

other: Please see attached tabels 3 and 5 for further details on individual animals. The animals of all dose groups showed apathy at least at one day of administration. The most clear symptoms in the animals of the two higher dose grous were decreasing of body w

Gross pathology:

Please refer to the attached tables 4 and 7 for further details.
In the surviving animals macroscopic pathological findings were not observed except for a slight haemorrhagic lung in the male animal No5 of the middle dose group. There were no gross lesions in the animals of the low dose group except for one of the died female animals with a slight lung emphysema. In the two higher dose groups the lungs of all died animals showed emphysema, in many cases with haemorrhagic regions. The mouth was agglutinated with saliva in all died animals of these dose groups. The contents of the stomach and small intestine showed that the animals did not ingest feed until death.

Any other information on results incl. tables

Table 1: Mortality

Dose group (mg/kg b.w.)	Male animals		Female animals
	Number of died animals of 5	Mortality in %	Number of died animals of 5	Mortality in %
2000	5	100	-	-
1500	2	40	-	-
1300	-	-	4	80
1150	-	-	1	20
1000	0	0	1	20
Results of probit analysis by E. Weber
Chiquadrat comparison: χ^2(0.95; DF), table: χ^2, calculated: Homogeneity			0.4 1.3 heterogeneous
Regression evaluation:	Y = a + bX Y: data in probit; X: data in log dose
Slope^*(b) Intercept(a) Degree of Freedom (DF) t-value(5% DF) Correction term g b is:			14.8 -44.1 1 12.7 62.6 valid
LD50(mg/kg b.w.) Confidence interval			1199 not available

^*Slope = Regression coefficient

Because of the all-or-none mortality response in male animals of the high or low dose group, it was not possible to calculate a precise LD50 value according to the probit analysis by E. Weber. However, from these data the LD50 value can be reasonably estimated to be about 1600mg/kg b.w.

The regression calculation was not valid in the female animals because the results havea too great variation arount the regression straight line. The LD50 value was calculated to be about 1200mg/kg b.w. without valid slope and condifdence interval.

A test continuation with a further dose group is not justified for ethical reasons because the result is unambiguous for the classification in accordance with the German "Chemical Act" (in each case 200=<LD50<2000 mg/kg b.w.).

Table 2: Time of Death

	Dose group [mg/kg b.w.]	Animal N^o	Time of Death Hours after administration
Males	2000	1, 5	> 5 hours < 22 hours
		2	approximately 29 hours
		4	approximately 30 hours
		3	> 30 hours < 46 hours
	1500	4	approximately 27 hours
		3	> 32 hours < 48 hours
Females	1300	5	> 8 hours < 24 hours
		2, 4	> 32 hours < 48 hours
		3	55 hours, moribundly killed
	1150	4	> 30 hours < 46 hours
	1000	4	> 30 hours < 46 hours

Applicant's summary and conclusion

Interpretation of results:

harmful

Remarks:

if swallowed Criteria used for interpretation of results: EU

Conclusions:

See "Overall remarks" for conclusion and interpretation of results.

Executive summary:

Acute oral toxicity of N-(2,2 -Dimethoxyethyl)acrylamide was tested in male and female Charles River Wistar rats. The test article was administered at the single doses of 2000, 1500 and 1000 mg/kg body weight (b.w.) in male animals and of 1300, 1150 and 1000 mg/kg b.w. in female animals by gavage (five male animals and five female animals per group).

All animals were examined for mortality, clinical signs and body weight gain. The pathological alterations of organs were examined in the died animals shortly after the death or in the surviving animals at the end of a 14-day observation period.

The LD₅₀ values were estimated to about 1600 mg/kg b.w. in male animals and to about 1200 mg/kg b.w. in female animals.

Mortalities occurred in the night after administration until the second night after administration.

The most clear clinical symptoms were apathy, decreasing of body weight in animals which died during the course of investigation, tremor, bloody rhinorrhea and increased salivation. All these symptoms were dose dependently increased.

The body weight gain was not affected in the animals of the low dose groups, but it was slightly delayed in the surviving animals of the two higher dose groups.

The most clear macroscopic pathological findings in all died animals were pulmonary emphysema, in many cases with hemorrhagic regions and the mouths were agglutinated with saliva.

The pulmonary emphysema could be seen as cause of death. The tremor is a sign of the central action of the test substance.

In accordance with the results of this test and in compliance with the criteria of Annex IV of Council Directive 93/21/EEC of 27th April 1993 N-(2,2 -Dimethoxyethyl)acrylamide must be classified as harmful (symbol of danger:Xn) and must be labelled with R22 “Harmful if swallowed”.