1H-Imidazole-1-propanenitrile

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(only 2-aminoanthracene as positve control with metabolic activation)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Imidazole-1-propionitrile(8CI);1-(2-Cyanoethyl)imidazole

EC Number:

607-301-1

Cas Number:

23996-53-4

Molecular formula:

C6H7N3

IUPAC Name:

Imidazole-1-propionitrile(8CI);1-(2-Cyanoethyl)imidazole

Details on test material:

- Name of test substance (as cited in study report): Cyanethylimidazol
- Analytical purity: > 95%
- Batch No.: K 94/1
- Substance No.: 96/124
- Physical state: liquid (brownish, clear)
- Storage: refrigerator

Method

Target gene:

His- and Trp-operon

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplement post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.

Test concentrations with justification for top dose:

20, 100, 500, 2500 and 5000 µg/plate (experiment 1 and 2)

Vehicle / solvent:

water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: (+ S9): 2-aminoanthracene (2AA; all strains); (- S9): N-methyl-N'-nitro-N-nitroso-guanidine (MNNG; TA 100 and TA 1535), 4-nitro-o-phenylendiamine (NOPD; TA 98), 9-aminoacridine (AAC; TA 1537), N-ethyl-N'-nitro-N-nitroso-guanidine (ENNG; E. coli WP2 uvrA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) (experiment 1)

DURATION
- Exposure duration: 48 - 72 h at 37 °C

METHOD OF APPLICATION: preincubation (experiment 2)

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h at 37 °C

NUMBER OF REPLICATIONS: 3 plates per concentration per experiment

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of revertant colonies; clearing or diminution of the background lawn; degree of survival of treated cultures

Evaluation criteria:

In general, a substance has to fulfill the following criteria to be characterized as positive in the bacterial tests:
- doubling of the spontaneous mutation rate (compared to control)
- dose-response realtionship
- reproducibility of the results.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitation occurred up to a concentration of 5000 µg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Test results of experiment 1 (plate incorporation).

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	E. coli WP2 uvrA	TA98	TA1537
–	0	119 ± 10	18 ± 1	33 ± 8	25 ± 4	8 ± 1
–	20	122 ± 3	15 ± 1	36 ± 3	30 ± 1	9 ± 1
–	100	121 ± 11	20 ± 4	39 ± 1	20 ± 2	8 ± 3
–	500	134 ± 6	18 ± 2	37 ± 8	24 ± 4	9 ± 1
–	2500	129 ± 8	18 ± 3	36 ± 6	27 ± 5	10 ± 2
–	5000	128 ± 22	15 ± 3	47 ± 3	26 ± 5	6 ± 4
Positive controls, –S9	Name	MNNG	MNNG	ENNG	NOPD	AAC
	Concentrations (μg/plate)	5	5	10	10	100
	Mean No. of colonies/plate (average of 3 ± SD)	1214 ± 154	1096 ± 139	560 ± 55	692 ± 113	777 ± 131
+	0	126 ± 9	19 ± 3	34 ± 6	39 ± 3	9 ± 1
+	20	142 ± 7	16 ± 2	40 ± 6	36 ± 4	12 ± 1
+	100	135 ± 15	15 ± 5	38 ± 1	41 ± 1	12 ± 2
+	500	112 ± 9	16 ± 4	41 ± 2	45 ± 4	10 ± 1
+	2500	115 ± 1	17 ± 2	40 ± 10	40 ± 2	9 ± 2
+	5000	123 ± 7	14 ± 7	45 ± 3	27 ± 7	7 ± 1
Positive controls, +S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	2.5	60	2.5	2.5
	Mean No. of colonies/plate (average of 3 ± SD)	1531 ± 80	1033 ± 45	262 ± 49	864 ± 102	224 ± 12

ENNG = N-ethyl-N-nitro-N-nitrosoguanidine

MNNG = N-methyl-N-nitro-N-nitrosoguanidine

NOPD = 4-nitroquinoline-N-oxide

AAC = 9-aminoacridine

2AA = 2-Aminoanthracene

 

Table 2. Test results of experiment 2 (preincubation test).

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	E. coli WP2 uvrA	TA98	TA1537
–	0	128 ± 5	21 ± 2	35 ± 5	29 ± 6	13 ± 2
–	20	105 ± 6	16 ± 1	33 ± 5	28 ± 5	8 ± 3
–	100	100 ± 5	19 ± 2	29 ± 2	28 ±2	11 ± 4
–	500	113 v 11	18 ± 2	34 ± 5	26 ± 4	10 ± 2
–	2500	109 ± 12	16 ± 2	28 ± 3	29 ± 8	11 ± 2
–	5000	113 ± 9	18 ± 2	30 ± 1	28 ± 7	10 ± 3
Positive controls, –S9	Name	MNNG	MNNG	ENNG	NOPD	AAC
	Concentrations (μg/plate)	5	5	10	10	100
	Mean No. of colonies/plate (average of 3 ± SD)	1132 ± 25	1200 ± 54	530 ± 19	1466 ± 25	457 ± 49
+	0	138 ± 4	21 ± 2	45 ± 5	48 ± 4	13 ± 1
+	20	107 ± 7	16 ± 3	37 ± 4	50 ± 3	15 ± 1
+	100	110 ± 9	15 ± 2	41 ± 5	45 ± 6	15 ± 3
+	500	114 ± 9	12 ± 3	38 ± 1	53 ± 4	11 ± 3
+	2500	111 ± 11	16 ± 1	48 ± 1	45 ± 7	10 ± 3
+	5000	106 ± 7	17 ± 3	42 ± 5	49 ± 6	11 ± 2
Positive controls, +S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	2.5	60	2.5	2.5
	Mean No. of colonies/plate (average of 3 ± SD)	1097 ± 36	124 ± 7	204 ± 7	925 ± 40	112 ± 8

ENNG = N-ethyl-N-nitro-N-nitrosoguanidine

MNNG = N-methyl-N-nitro-N-nitrosoguanidine

NOPD = 4-nitroquinoline-N-oxide

AAC = 9-aminoacridine

2AA = 2-Aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative