1,3-divinylimidazolidin-2-one

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993-06-02

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP compliant and guideline similar study. Only 4 strains were tested.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

, 4 instead 5 strains tested

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,3-divinylimidazolidin-2-one

EC Number:

237-457-2

EC Name:

1,3-divinylimidazolidin-2-one

Cas Number:

13811-50-2

Molecular formula:

C7H10N2O

IUPAC Name:

1,3-diethenylimidazolidin-2-one

Test material form:

solid: flakes

Details on test material:

- Name of the test substance: N,N'-Divinylimidazolidon
- Batch no.: 26300/9
- Analytical purity: 96.3 %
- Date of manufacturing: 1993-01-20
- Appearance: Colorless to yellow flakes
- Storage: Refrigerator

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

other: TA 1535 and TA 100 used to detect base pair substitutions; TA 1537 and TA 98 are strains for the detection of frameshift mutagens (+1 mutant his C 3076 in the case of TA 1537 and the +2 type his D 3052 in the case of TA 98)

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat (male) liver S-9 mix

Test concentrations with justification for top dose:

20, 100, 500, 2500, 5000 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with metabolic activation: 2-aminoanthracene (all strains); without metabolic activation: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG for TA 100 and TA 1535), 4-nitro-o-phenylendiamine (NPD for TA 98) and 9-aminoacridine chloride monohydrate (ACC, TA 1537)

Details on test system and experimental conditions:

METHOD OF APPLICATION: In agar (plate incorporation) and preincubation

Standard plate test:

2 mL portions of soft agar which consists of 100 mL agar and 10 mL amino acid solution are kept in a water bath at 45°C, the remaining components were added in the following order:
0.1 mL test solution or vehicle
0.1 mL bacteria] suspension
0.5 mL S-9 mix (in tests with metabolic activation)
or
0.5 mL phosphate buffer (in tests without metabolic activation )

After mixing, the samples were poured onto Vogel-Bonner agar plates (minimal glucose agar plates) within approx. 30 seconds.


Preincubation test:

0.1 mL test solution or vehicle, 0 .1 mL bacterial suspension and 0.5 mL S-9 mix are incubated at 37°C for the duration of 20 minutes. Subsequently, 2 mL of soft agar was added and, after mixing, the samples were poured onto the Vogel-Bonner agar plates within approx. 30 seconds.
After incubation at 37°C for 48 hours in the dark, the bacterial colonies (his revertants) were counted.

DURATION
- Exposure duration: 20 min
- Expression time: 48 hours


NUMBER OF REPLICATIONS: 3 test plates per dose or per control

DETERMINATION OF CYTOTOXICITY
- Method: Relative total growth

OTHER EXAMINATIONS:
Strains are checked for the following characteristics at regular intervals : deep rough character (rfa) ; UV sensitivity (A uvrB) ; ampicillin resistance (R factor plasmid).
Histidine auxotrophy is automatically checked in each experiment via the spontaneous rate.

Evaluation criteria:

In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: A weakly bacteriotoxic effect was observed only with the strains TA 1535 and TA 100 (details see below).

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

An increase in the number of his+ revertants was not observed both in the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system.


TEST-SPECIFIC CONFOUNDING FACTORS

- Precipitation: No test substance precipitation was found.


ADDITIONAL INFORMATION ON CYTOTOXICITY:

A slight decrease in the number of his(+) revertants was observed in the standard plate test and preincubation assay using TA 100 depending on the test conditions at doses >= 500 µg/plate.
A weakly bacteriotoxic effect was also found in the preincubation test without S-9 mix at 5000 µg/plate with TA 1535.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Number of revertants per strain (mean from three plates), Standard Plate Test

 

	Strain TA 1535		Strain TA 1537		Strain TA 98		Strain TA 100
Conc. [µg/plate]	-S9 mix	+S9 mix	-S9 mix	+S9 mix	-S9 mix	+S9 mix	-S9 mix	+S9 mix
Positive control*	90	785	317	88	725	561	723	478
Negative control	20	19	10	12	34	44	114	129
20	23	19	8	11	29	39	113	110
100	25	20	11	12	38	27	110	108
500	20	19	9	10	32	33	120	91
2500	24	15	9	18	34	59	86	80
5000	23	18	8	14	29	42	74	75

*without S9 mix: MNNG (5 µg/plate): TA1535, TA 100; AAC (100 µg/plate): TA 1537; NPD (10 µg/plate): TA98

*with S9 mix: 2-AA (2.5 µg/plate): TA1535, TA 100; TA 1537, TA 98

 

 

Table 2: Number of revertants per strain (mean from three plates), Preincubation Test

 

	Strain TA 1535		Strain TA 100		Strain TA 1537		TA 98
Conc. [µg/plate]	-S9 mix	+S9 mix	-S9 mix	+S9 mix	-S9 mix	+S9 mix	-S9 mix	+S9 mix
Positive control*	827	93	1088	487	428	79	928	342
Negative control	19	13	107	108	9	10	28	43
20	10	14	96	113	8	10	31	36
100	11	15	99	123	6	8	33	38
500	13	16	64	100	7	9	28	36
2500	14	12	69	100	12	10	33	35
5000	9	10	17	76	9	12	32	42

*without S9 mix: MNNG (5 µg/plate): TA1535, TA 100; AAC (100 µg/plate): TA 1537; NPD (10 µg/plate): TA98

*with S9 mix: 2-AA (2.5 µg/plate): TA1535, TA 100; TA 1537, TA 98

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative