2-imidazolidone

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:

1991

Report date:

1991

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-imidazolidone

EC Number:

204-436-4

EC Name:

2-imidazolidone

Cas Number:

120-93-4

Molecular formula:

C3H6N2O

IUPAC Name:

2-imidazolidone

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 74-7718
- Test substance No.: 89/91

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

Method

Target gene:

his

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254

Test concentrations with justification for top dose:

- first test: 20, 100, 500, 2500 and 5000 µg/plate (standard plate test; all test strains)
- second test: 200, 400, 600, 800 and 1000 µg/plate (standard plate test; test strain TA98)
- third test: 20, 100, 500, 2500 and 5000 µg/plate (preincubation test; all test strains)
- fourth test: 4, 20, 100, 500 and 2500 µg/plate (preincubation test; test strain TA98)

Vehicle / solvent:

aqua dest.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: - S9: aminoanthracene (2-AA): all test strains; + S9: methyl-N-nitro-N-nitrosoguanidine (MNNG): TA100 and TA1535, 4-nitro-o-phenylendiamine (NPD): TA98 and 9-aminoacridine chloride monohydrate (AAC): TA1537

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
Plate incorcoration method:
- Exposure duration: ca. 48-72 hours at 37 °C in the dark
Preincubation method:
- Preincubation period: 20 minutes at 37 °C
- Exposure duration: ca. 48-72 hours at 37 °C in the dark

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

DETERMINATION OF CYTOTOXICITY
- Decrease in the number of revertants
- Reduced his- or trp- background growth
- Reduction in the titer

Evaluation criteria:

In order to characterized a substance as positive in the Ames test it 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

Key result

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:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Toxicity:
No bacteriotoxic effect (reduced his- background growth) was observed.

Solubility:
Complete solubility of test substance in aqua dest.

Any other information on results incl. tables

Table 1: Standard Plate Test (TA 98)

Dose [µg/plate]	TA 98 Mean ± SD
Without S9 mix
Negative control (aqua dest)	28 ± 5
200	33 ± 12
400	26 ± 4
600	24 ± 5
800	20 ± 2
1000	26 ± 4
10 NPD	1118 ± 125
With S9 mix
Negative control (aqua dest)	44 ± 4
200	38 ± 4
400	40 ± 3
600	36 ± 1
800	39 ± 3
1000	30 ± 11
10 2-AA	1850 ± 26

Table 2: Standard Plate Test (TA 98, TA 100, TA 1535, TA 1537)

Dose [µg/plate]	TA 98 Mean ± SD	TA 100 Mean ± SD	TA 1535 Mean ± SD	TA1537 Mean ± SD
Without S9 mix
Negative control (aqua dest)	28 ± 2	140 ± 5	19 ± 6	15 ± 4
20	33 ± 5	129 ± 8	24 ± 3	10 ± 4
100	33 ± 3	131 ± 27	19 ± 1	15 ± 4
500	38 ± 11	99 ± 8	22 ± 5	13 ± 5
2500	26 ± 6	114 ± 6	22 ± 4	10 ± 5
5000	23 ± 7	139 ± 5	23 ± 4	10 ± 3
10 NPD	922 ± 88
100 AAC				430 ± 91
5 MNNG		1347 ± 43	1349 ± 273
With S9 mix
Negative control (aqua dest)	41 ± 5	122 ± 13	20 ± 11	14 ± 2
20	57 ± 5	139 ± 6	21 ± 4	15 ± 5
100	54 ± 12	126 ± 4	18 ± 2	15 ± 7
500	62 ± 23	137 ± 16	20 ± 4	21 ± 1
2500	36 ± 9	140 ± 45	19 ± 4	20 ± 8
5000	36 ± 10	117 ± 30	18 ± 1	16 ± 3
10 2-AA	785 ± 62	334 ± 47	379 ± 53	96 ± 15

Table 3: Preincubation Test (TA 98)

Dose [µg/plate]	TA 98 Mean ± SD
Without S9 mix
Negative control (aqua dest)	26 ± 2
4	23 ± 2
20	26 ± 2
100	21 ± 2
500	27 ± 1
2500	26 ± 2
10 NPD	1120 ± 98
With S9 mix
Negative control (aqua dest)	41 ± 4
4	39 ± 6
20	34 ± 13
100	35 ± 13
500	38 ± 6
2500	33 ± 4
10 2-AA	1247 ± 87

Table 4: Preincubation Test (TA 98, TA 100, TA 1535, TA 1537)

Dose [µg/plate]	TA 98 Mean ± SD	TA 100 Mean ± SD	TA 1535 Mean ± SD	TA 1537 Mean ± SD
Without S9 mix
Negative control (aqua dest)	24 ± 4	104 ± 11	14 ± 3	12 ± 2
20	39 ± 2	117 ± 16	15 ± 3	12 ± 2
100	36 ± 3	95 ± 15	16 ± 2	14 ± 1
500	38 ± 6	95 ± 5	16 ± 6	9 ± 4
2500	39 ± 9	115 ± 13	19 ± 3	13 ± 2
5000	31 ± 2	118 ± 11	14 ± 2	11 ± 2
10 NPD	1320 ± 131
100 AAC				313 ± 36
5 MNNG		1343 ± 81	1317 ± 6
With S9 mix
Negative control (aqua dest)	35 ± 3	113 ± 19	17 ± 6	12 ± 1
20	59 ± 20	126 ± 12	18 ± 2	14 ± 2
100	51 ± 10	148 ± 18	19 ± 1	11 ± 1
500	49 ± 4	129 ± 18	14 ± 2	14 ± 5
2500	48 ± 5	126 ± 21	17 ± 2	20 ± 3
5000	46 ± 3	135 ± 10	17 ± 2	19 ± 1
10 2-AA	865 ± 241	886 ± 63	167 ± 11	141 ± 22

2 -AA: 2 -aminoanthracene

MNNG: N-methyl-N-nitro-N-nitroso-guanidine

NPD: 4 -nitro-o-phenylendiamine

AAC: 9 -aminoacridine chloride monohydrate