N'-[3-(dimethylamino)propyl]-N,N-dimethylpropane-1,3-diamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

N'-[3-(dimethylamino)propyl]-N,N-dimethylpropane-1,3-diamine was negative in an Ames test, test guideline OECD 471, GLP (BASF AG 1998).

 

 

Link to relevant study records

Reference

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:

other: Guideline study with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

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

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Target gene:

his operon and trp operon

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

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

Test concentrations with justification for top dose:

Experiment 1: standard plate test 1: 0; 20; 100; 500; 2500 and 5000 µg/plate (with and without S9-mix); all strains
Experiment 2: standard plate test 2: 2000, 3000, 4000, 5000, 6000 µg/plate (with and without S9-mix); (TA100, E.coli WP2 uvrA)
Experiment 3: preincubation test : 0; 20; 100; 500; 2500 and 5000 µg/plate (with and without S9-mix); all strains

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Untreated negative controls:

yes

Remarks:

sterility control

Negative solvent / vehicle controls:

yes

Remarks:

water control

True negative controls:

no

Positive controls:

yes

Positive control substance:

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

Remarks:

2-AA: 2 µg (TA100, TA 98, TA 1537, TA 1535), 60 µg (E. coli WP2 uvrA); MNNG: 5 µg; NOPD: 10 µg; AAC: 100 µg; ENNG: 10 µg; solvent for all positive control substances: DMSO

Details on test system and experimental conditions:

METHOD OF APPLICATION: standard plate test (first experiment), preincubation test (second experiment)

DURATION
- Preincubation period: 20 min (only preincubation test)
- Exposure duration: 48 - 72 h (in the dark, 37 °C)

NUMBER OF REPLICATIONS: 3 plates/concentration/experiment; 3 3experiments

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of revertants or a clearing of the bacterial background lawn, reduction of the titer.
The titer is generally determined only in the experimental parts with S9 mix both for the negative controls (vehicle only) and for the two highest doses in all experiments.

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 doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

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:

cytotoxicity

Remarks:

A weak bacteriotoxic effect in the standard plate test at 6000 µg/plate (TA 100). In the preincubation assay bacteriotoxicity was found depending on the strain and test conditions at doses >= 500 µg/plate.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A weak bacteriotoxic effect in the standard plate test at 6000 µg/plate. In the preincubation assay bacteriotoxicity was found at doses >= 2500 µg/plate.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no test substance precipitation was found

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

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	WP2 uvrA	TA98	TA1537
–	0	116 ± 13	14 ± 2	30 ± 1	27 ± 5	9 ± 2
–	20	130 ± 32	13 ± 3	39 ± 3	25 ± 7	8 ± 2
–	100	153 ± 14	16 ± 5	41 ± 7	32 ± 5	8 ± 1
–	500	161 ± 23	20 ± 4	37 ± 8	22 ± 7	10 ± 2
–	2500	153 ± 13	18 ± 2	48 ± 4	25 ± 3	9 ± 3
–	5000	179 ± 3	21 ± 9	44 ± 7	27 ± 8	9 ± 2
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)	1223 ± 268	1070 ± 278	698 ± 36	982 ± 16	575 ± 6
+	0	119 ± 13	15 ± 3	46 ± 5	42 ± 4	9 ± 2
+	20	171 ± 24	16 ± 5	56 ± 5	40 ± 8	11 ± 0
+	100	143 ± 23	15 ± 5	39 ± 4	40 ± 13	10 ± 3
+	500	138 ± 22	19 ± 5	44 ± 6	43 ± 6	12 ± 3
+	2500	154 ± 10	22 ± 3	54 ± 13	40 ± 3	7 ± 1
+	5000	147 ± 17	21 ± 3	57 ± 7	33 ± 4	9 ± 3
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)	1297 ± 57	112 ± 3	327 ± 37	577 ± 33	214 ± 21

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

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

NOPD = 4-nitro-o-phenylendiamine

AAC = 9-aminoacridine

2AA = 2-Aminoanthracene

B = reduced background growth

Table 2: Test results of experiment 2 (plate incorporation)

 

With or without S9-Mix	Test substance concentration (μg/plate)
		TA 100	WP2 uvrA
–	0	117 ± 5	35 ± 2
–	2000	105 ± 5	38 ± 3
–	3000	120 ± 13	41 ± 2
–	4000	108 ± 11	38 ± 7
–	5000	90 ± 10	45 ± 4
–	6000	148 ± 20	31 ± 9
Positive controls, –S9	Name	MNNG	ENNG
	Concentrations (μg/plate)	5	10
	Mean No. of colonies/plate (average of 3 ± SD)	1307 ± 138	505 ± 6
+	0	126 ± 28	36 ± 6
+	2000	136 ± 11	32 ± 3
+	3000	142 ± 5	32 ± 4
+	4000	119 ± 19	36 ± 4
+	5000	97 ± 11	24 ± 3
+	6000	0B	0B
Positive controls, +S9	Name	2AA	2AA
	Concentrations (μg/plate)	2.5	60
	Mean No. of colonies/plate (average of 3 ± SD)	1182 ± 66	220 ± 34

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

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

NOPD = 4-nitro-o-phenylendiamine

AAC = 9-aminoacridine

2AA = 2-Aminoanthracene

B = reduced background growth

 

 

Table 3: Test results of experiment 3 (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	WP2 uvrA	TA98	TA1537
–	0	141 ± 10	22 ± 4	33 ± 4	31 ± 3	8 ± 2
–	20	117 ± 4	18 ± 1	36 ± 2	28 ± 7	9 ± 2
–	100	113 ± 13	19 ± 1	34 ± 4	30 ± 2	8 ± 1
–	500	94 ± 8	7 ± 2	34 ± 5	16 ± 2	8 ± 1
–	2500	0B	0B	14 ± 3	0B	0B
–	5000	0B	0B	0B	0B	0B
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)	1104 ± 47	789 ± 44	521 ± 33	1087 ± 70	408 ± 42
+	0	138 ± 10	23 ± 0	41 ± 7	46 ± 4	9 ± 1
+	20	124 ± 10	20 ± 3	40 ± 3	41 ± 2	11 ± 3
+	100	103 ± 1	21 ± 3	40 ± 2	44 ± 9	13 ± 6
+	500	88 ± 8	21 ± 3	35 ± 3	36 ± 9	11 ± 5
+	2500	102 ± 5	13 ± 3	50 ± 4	30 ± 7	8 ± 2
+	5000	43 ± 9B	0B	18 ± 2	0B	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)	983 ± 31	121 ± 9	227 ± 14	574 ± 54	103 ± 11

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

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

NOPD = 4-nitro-o-phenylendiamine

AAC = 9-aminoacridine

2AA = 2-Aminoanthracene

B = reduced background growth

Conclusions:

Interpretation of results (migrated information):
negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Bacterial reverse mutation assay:

BASF (1998) performed an Ames (plate incorporation assay and preincubation test) test with S. typhimurium strains TA1535, TA1537, TA98 and TA100 and E. coli WP2 with and without metabolic activation in presence of Bis (3-dimethylaminopropyl) amin (Bis-DMAPA).

Following test concentrations were applied: 0, 20, 100, 500, 2500 and 5000 µg/plate (in triplicate) with and without metabolic activation (S9 -mix) using the standard plate incorporation assay for all strains, and 2000, 3000, 4000, 5000 and 6000 µg/plate (with and without S9 -mix) for S. typhimurium strain TA100 and E. coli WP2 uvrA. Solvent control, negative control and positive controls were run in triplicate. Following cytotoxic effects were observed in Salmonella strains: A weak bacteriotoxic effect in the standard plate test at 6000 µg/plate (TA 100). In the preincubation assay bacteriotoxicity was found depending on the strain and test conditions at doses >= 500 µg/plate. Following cytotoxic effects were observed in E. coli WP2 uvrA: toxic effect in the standard plate test at 6000 µg/plate. In the preincubation assay bacteriotoxicity was found at doses >= 2500 µg/plate. Solvent, negative and positive controls were valid.

 

Justification for classification or non-classification

Classification,Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for genetic toxicity under Regulation (EC) No. 1272/2008, as amended for the third time in Directive (EC 618/2012).