N-methyl-N-vinylacetamide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-methyl-N-vinylacetamide

EC Number:

221-698-5

EC Name:

N-methyl-N-vinylacetamide

Cas Number:

3195-78-6

Molecular formula:

C5H9NO

IUPAC Name:

N-ethenyl-N-methylacetamide

Specific details on test material used for the study:

Batch no: 4043/97-17

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-induced rat liver S-9 mix

Test concentrations with justification for top dose:

20, 100, 500, 2500 and 5000 ug/plate (concentrations for Standard Plate Test and Preincubation Test)

Vehicle / solvent:

Due to the good solubility of the test substance in water, water was selected as the vehicle.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with metabolic activation: 2-aminoanthracene; without metabolic activation: N-methyl-N'-nitro-N-nitrosoguanidine, 4-nitro-o-phenylendiamine, 9-aminoacridine and 4-nitroquinoline-N-oxide

Remarks:

Parallel with each experiment, negative controls are carried out in order to check for possible contaminants (sterility control) and to determine the spontaneous mutation rate (vehicle control).

Details on test system and experimental conditions:

METHOD OF APPLICATION: 1st Experiment: in agar (plate incorporation), 2nd Experiment: preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48-72 hours

NUMBER OF REPLICATIONS: 3 test plates per dose or per control in the 1st and 2nd experiment, each

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of background lawn (= reduced his- or trp- background growth), reduction in the titer

OTHER: Determination of solubility
As long as precipitation does not interfere with the colony scoring, 5 mg/plate is generally selected and analyzed (in cases of nontoxic compounds) as the maximum dose at least in the 1st experiment even in the case of relatively insoluble test compounds to detect possible mutagenic impurities. Furthermore, doses > 5 mg/plate might also be tested in repeat experiments for further clarification/substantiation.

Evaluation criteria:

Acceptance criteria:
Generally, the experiment is to be considered valid if the following criteria are met:
- The number of revertant colonies in the negative controls was within the normal range of the historical control data for each tester strain.
- The sterility controls revealed no indication of bacterial contamination.
- The positive control articles both with and withaut S-9 mix induced a significant inerease in the number of revertant colonies withinthe range of the historical control data.
- The titer of viable bacteria was >10E9/mL.

Evaluation criteria:
The test chemical is considered positive in this assay if the following criteria are met:
- A dose-related and reproducible increase in the number~of revertant colonies, i.e. about daubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.
A test substance is generally considered nonmutagenic in this test if:
- The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in twa experiments carried aut independently af each other.

Results and discussion

Test results

Key result

Species / strain:

bacteria, other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: A slight decrease in the number of revertants was occasionally observed.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No precipitation of the test substance was found.

Any other information on results incl. tables

Standard plate test

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium and E. coli
	TA1535	TA100	TA1537	TA98	WP2 uvrA
	Results without S9
Water	18 ± 1	106 ± 5	9 ± 2	25 ± 1	25 ± 1
20	19 ± 2	116 ± 14	7 ± 2	22 ± 6	22 ± 2
100	15 ± 2	98 ± 2	7 ± 1	25 ± 2	25 ± 6
500	16 ± 0	83 ± 8	5 ± 1	27 ± 5	25 ± 3
2500	15 ± 3	81 ± 1	5 ± 1	21 ± 3	26 ± 3
5000	13 ± 3	92 ± 5	6 ± 1	22 ± 4	23 ± 3
MNNG (5.0)	721 ± 26	755 ± 23
AAC (100)			771 ± 16
NOPD (10)				689 ± 70
4-NQO (5)					895 ± 58
	Results with S9
Water	19 ± 1	141 ± 12	10 ± 2	38 ± 3	37 ± 2
20	16 ± 2	122 ± 7	11 ± 3	39 ± 3	28 ± 2
100	16 ± 2	145 ± 15	8 ± 1	31 ± 2	31 ± 3
500	15 ± 2	142 ± 23	6 ± 1	26 ± 4	26 ± 6
2500	10 ± 3	124 ± 5	7 ± 1	27 ± 5	25 ± 5
5000	8 ± 1	162 ± 14	7 ± 2	29 ± 4	22 ± 3
2-AA (2.5)	223 ± 6	1460 ± 169	165 ± 11	914 ± 33
2-AA (60.0)					223 ± 17

 

Pre incubation test

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium and E. coli
	TA1535	TA100	TA1537	TA98	WP2 uvrA
	Results without S9
Water	20 ± 2	131 ± 5	10 ± 2	27 ± 3	26 ± 3
20	18 ± 2	135 ± 11	11 ± 2	27 ± 2	24 ± 3
100	17 ± 2	120 ± 11	8 ± 1	23 ± 3	31 ± 4
500	18 ± 2	124 ± 4	6 ± 2	20 ± 3	23 ± 4
2500	15 ± 3	125 ± 5	6 ± 2	20 ± 3	23 ± 1
5000	16 ± 3	124 ± 9	6 ± 1	20 ± 2	22 ± 2
MNNG (5.0)	1133 ± 235	1105 ± 32
AAC (100)			660 ± 35
NOPD (10)				989 ± 13
4-NQO (5)					802 ± 35
	Results with S9
Water	21 ± 3	139 ± 14	12 ± 3	38 ± 1	36 ± 4
20	18 ± 2	133 ± 9	12 ± 5	39 ± 5	27 ± 3
100	18 ± 2	145 ± 9	9 ± 1	34 ± 3	29 ± 4
500	17 ± 1	145 ± 7	12 ± 1	34 ± 6	33 ± 1
2500	17 ± 2	148 ± 9	8 ± 3	34 ± 2	41 ± 2
5000	16 ± 3	139 ± 4	5 ± 1	27 ± 3	44 ± 5
2-AA (2.5)	113 ± 14	681 ± 18	148 ± 12	870 ± 62
2-AA (60.0)					182 ± 14