3-diethylaminopropiononitrile

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP (only 2-aminoanthracene as positive control with metabolic activation)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted 1997)

Deviations:

yes

Remarks:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

and OECD Guidline 472 (adopted 1983)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-diethylaminopropiononitrile

EC Number:

226-323-9

EC Name:

3-diethylaminopropiononitrile

Cas Number:

5351-04-2

Molecular formula:

C7H14N2

IUPAC Name:

3-(diethylamino)propanenitrile

Details on test material:

- Name of test substance (as cited in study report): N,N-Diethylaminopropionitril
- Analytical purity: 99.6%
- Test substance number: 96/119
- Batch numer: K 80/1 Hauptlauf
- Physical state: Colourless liquid
- Storage conditions: refrigerator

Method

Target gene:

His- and Trp-operon

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9 (S9 mix: 1 part S9 and 9 parts S9 supplement with cofactors)

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): MNNG: TA 100 and TA 1535; 4-nitro-o-henylendiamine (NOPD): TA 98; 9-aminoacridine (AAC): TA 1537; ENNG: E. coli WP2 uvrA; (+S9): 2-aminoantracene (2AA): TA 100, TA 98, TA 1537, TA 1535 and E. coli WP2 uvrA

Remarks:

MNNG = N-methyl-N'-nitro-N-nitroso-guanidine; ENNG = N-ethylen-N'-nitro-N-nitroso-guanidin

Details on test system and experimental conditions:

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

DURATION
- Exposure duration: 48 - 72 h

METHOD OF APPLICATION: preincubation: Experiment 2

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: 3 plates/dose/experiment

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

Evaluation criteria:

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

Results and discussion

Test results

Species / strain:

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

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:

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	150 ± 12	14 ± 5	35 ± 4	39 ± 2	11 ± 4
–	20	135 ± 14	18 ± 4	30 ± 3	22 ± 3	10 ± 2
–	100	162 ± 10	17 ± 4	31 ± 3	37 ± 6	7 ± 1
–	500	143 ± 13	14 ± 3	35 ± 6	35 ± 13	9 ± 1
–	2500	131 ± 22	15 ± 1	36 ± 5	23 ± 3	8 ± 3
–	5000	138 ± 9	17 ± 1	37 ± 3	33 ± 7	10 ± 2
Positive controls, –S9	Name	MNNG	MNNG	ENNG	4NOPD	AAC
	Concentrations (μg/plate)	10	5	10	10	100
	Mean No. of colonies/plate (average of 3 ± SD)	1615 ± 81	748 ± 32	807 ± 61	1157 ± 91	267 ± 41
+	0	126 ± 2	22 ± 4	46 ± 10	46 ± 11	11 ± 5
+	20	142 ± 11	14 ± 3	34 ± 12	44 ± 10	8 ± 3
+	100	142 ± 10	22 ± 4	38 ± 6	57 ± 3	14 ± 3
+	500	153 ± 9	19 ± 5	40 ± 9	39 ± 5	10 ± 5
+	2500	145 ± 13	18 ± 2	44 ± 5	52 ± 7	10 ± 2
+	5000	141 ± 21	23 ± 6	34 ± 6	48 ± 6	9 ± 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)	1189 ± 116	142 ± 5	345 ± 12	1578 ± 32	126 ± 17

MNNG = N-methyl-N'-nitro-N-nitroso-guanidine

ENNG = N-ethylen-N'-nitro-N-nitroso-guanidine

4NOPD = 4-nitro-o-phenylene-diamine

AAC = 9-aminoacridine

2AA = 2-aminoanthracene

Table 2. Test results of experiment 2 (preincubation)

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	129 ± 24	16 ± 2	31 ± 5	29 ± 4	8 ± 4
–	20	137 ± 24	19 ± 2	28 ± 2	30 ± 6	8 ± 1
–	100	139 ± 24	17 ± 1	30 ± 2	30 ± 5	9 ± 3
–	500	135 ± 19	13 ± 3	33 ± 3	23 ± 2	7 ± 2
–	2500	143 ± 9	11 ± 3	28 ± 2	20 ± 3	8 ± 2
–	5000	146 ± 26	11 ± 2	26 ± 3	22 ± 3	8 ± 2
Positive controls, –S9	Name	MNNG	MNNG	ENNG	4NOPD	AAC
	Concentrations (μg/plate)	5	5	10	10	100
	Mean No. of colonies/plate (average of 3 ± SD)	1815 ± 93	1077 ± 43	610 ± 72	1484 ± 323	644 ± 135
+	0	143 ± 10	21 ± 4	39 ± 4	51 ± 4	9 ± 2
+	20	109 ± 6	15 ± 2	38 ± 7	38 ± 2	9 ± 2
+	100	138 ± 10	15 ± 2	32 ± 5	37 ± 1	8 ± 3
+	500	135 ± 27	11 ± 1	35 ± 3	34 ± 3	7 ± 2
+	2500	115 ± 6	11 ± 2	31 ± 8	36 ± 3	9 ± 4
+	5000	124 ± 17	9 ± 2	29 ± 3	29 ± 1	7 ± 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)	1017 ± 2	113 ± 19	242 ± 38	448 ± 73	129 ± 25

MNNG = N-methyl-N'-nitro-N-nitroso-guanidine

ENNG = N-ethylen-N'-nitro-N-nitroso-guanidine

4NOPD = 4-nitro-o-phenylene-diamine

AAC = 9-aminoacridine

2AA = 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative