Bis(2-dimethylaminoethyl)(methyl)amine

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:

other: GLP guideline study (OECD 471)

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 1 substance as positive control with S9-mix)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis(2-dimethylaminoethyl)(methyl)amine

EC Number:

221-201-1

EC Name:

Bis(2-dimethylaminoethyl)(methyl)amine

Cas Number:

3030-47-5

Molecular formula:

C9H23N3

IUPAC Name:

(2-{[2-(dimethylamino)ethyl](methyl)amino}ethyl)dimethylamine

Details on test material:

- Name of test material (as cited in study report): N,N,N',N",N"-Pentamethyldiethylentriamin
- Physical state: liquid, colourless
- Analytical purity: 99.3 %
- Lot/batch No.: Tank 12
- Storage condition of test material: Room temperature (N2 conditions)
- Other: substance manufacturing 1995-12; substance characterization was done prior to beginning of study (22-05-1998 to 1998-07-02; study was conducted from 1996-07-03 to 1996-07-10)

Method

Target gene:

his operon and trp operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

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:

First experiment: 0, 20, 100, 500, 2,500, 5,000 µg/plate
Second experiment: 0, 20, 100, 500, 2,500, 5,000 µg/plate

Vehicle / solvent:

water

Controls

Negative solvent / vehicle controls:

yes

Remarks:

(water)

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S9-mix: 2-AA (TA 98, TA 100, TA 1535, TA 1537, E. coli WP2uvrA2); without S9-mix: MNNG (TA 1535, TA 100); 4-NOPD (TA 98); AAC (TA 1537); ENNG (WP2 uvrA2)

Remarks:

2-Aminoanthracene (2-AA); N-methyl-N'-nitro-nitroso-guanidine (MNNG); 4-Nitro-o-phenylene-diamine (4-NOPD); 9-Aminoacridine (AAC); N-ethyl-N-nitro-N-nitrosoguanidine (ENNG)

Details on test system and experimental conditions:

Two tests were performed:

First test:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48- 72 hrs


NUMBER OF REPLICATIONS: triplicates/experiment

Second test
METHOD OF APPLICATION: preincubation

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

NUMBER OF REPLICATIONS: triplicates/experiment

DETERMINATION OF CYTOTOXICITY
- Method: other: determination of the titer, dimunition of background lawn, reduction of His(+)-revertants

Evaluation criteria:

A substance is to be considered as mutagenic when all following criteria are fulfilled:
1.) doubling of spontaneous mutation rate
2.) dose response-relationship
3.) reproducibility of results

Results and discussion

Test results

Test results 1

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

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Results for plate-incorporation test

Plate-incorporation test
With/ or without S9-mix	Test substance concentration [µg/plate]	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Salmonella typhymurium				E. coli
		Base-pair substitution type		Frameshift type
		TA 1535	TA 100	TA 1537	TA 98	WP2 uvrA
-	0	16 ± 2	149 ± 27	9 ± 0	33 ± 3	35 ± 10
-	20	12 ± 4	154 ± 32	9 ± 2	43 ± 11	44 ± 7
-	100	13 ± 2	190 ± 7	6 ± 1	33 ± 7	45 ± 10
-	500	12 ± 7	174 ± 10	10 ± 2	33 ± 4	41 ± 12
-	2500	13 ± 6	175 ± 35	8 ± 5	36 ± 7	49 ± 1
-	5000	9 ± 3	118 ± 6	10 ± 4	24 ± 6	41 ± 7
positive controls, -S9	Name	MNNG	MNNG	AAC	NOPD	ENNG
	Concentrations  [µg/plate]	5	5	100	10	10
	Mean no of colonies/plate (average of 3 ± SD)	760 ± 106	1252 ± 141	715 ± 22	958 ± 27	651 ± 46
+	0	16 ± 1	131 ± 25	9 ± 4	42 ± 1	54 ± 6
+	20	14 ± 3	124 ± 13	7 ± 2	50 ± 7	49 ± 5
+	100	12 ± 3	110 ± 6	8 ± 3	50 ± 10	53 ± 7
+	500	11 ± 1	104 ± 2	8 ± 4	50 ± 9	54 ± 9
+	2500	14 ± 6	112 ± 7	6 ± 2	51 ± 5	59 ± 11
+	5000	18 ± 3	129 ± 33	7 ± 4	36 ± 2	47 ± 3
positive controls, -S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations  [µg/plate]	2.5	2.5	2.5	2.5	60
	Mean no of colonies/plate (average of 3 ± SD)	114 ± 11	905 ± 77	114 ± 6	1182 ± 76	206 ± 13
MNNG = N-methyl-N'-nitro-N-nitrosoguanidine
AAC = 9-Aminoacridine
NOPD = 4-Nitro-o-phenylene-diamine
ENNG = N-ethyl-N-nitro-N-nitrosoguanidine
2AA= 2-Aminoanthracene

Table 2: Results for Pre-incubation test

Pre-incubationtest
With/ or without S9-mix	Test substance concentration [µg/plate]	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Salmonella typhymurium				E. coli
		Base-pair substitution type		Frameshift type
		TA 1535	TA 100	TA 1537	TA 98	WP2 uvrA
-	0	20 ± 3	137 ± 12	9 ± 1	29 ± 1	36 ± 5
-	20	20 ± 2	130 ± 7	9 ± 3	27 ± 4	32 ± 5
-	100	15 ± 2	126 ± 4	10 ± 2	25 ± 4	32 ± 3
-	500	14 ± 4	120 ± 7	9 ± 3	24 ± 2	30 ± 5
-	2500	13 ± 3	104 ± 13	6 ± 2	18 ± 4	25 ± 4
-	5000	12 ± 2	100 ± 19	6 ± 2	15 ± 5	23 ± 2
positive controls, -S9	Name	MNNG	MNNG	AAC	NOPD	ENNG
	Concentrations  [µg/plate]	5	5	100	10	10
	Mean no of colonies/plate (average of 3 ± SD)	1296 ± 74	943 ± 33	746 ± 71	1305 ± 30	825 ± 50
+	0	19 ± 2	136 ± 5	10 ± 1	43 ± 3	41 ± 7
+	20	16 ± 1	119 ± 3	9 ± 2	33 ± 5	37 ± 6
+	100	16 ± 1	99 ± 10	7 ± 1	31 ± 3	35 ± 5
+	500	14 ± 3	90 ± 11	5 ±1	27 ± 3	31 ± 4
+	2500	14 ± 2	99 ± 5	6 ± 1	23 ± 5	35 ± 1
+	5000	11 ± 2	96 ± 9	6 ± 1	26 ± 4	32 ± 2
positive controls, -S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations  [µg/plate]	2.5	2.5	2.5	2.5	60
	Mean no of colonies/plate (average of 3 ± SD)	129 ± 6	1360 ± 62	124 ± 3	1002 ± 9	177 ± 5
MNNG = N-methyl-N'-nitro-N-nitrosoguanidine
AAC = 9-Aminoacridine
NOPD = 4-Nitro-o-phenylene-diamine
ENNG = N-ethyl-N-nitro-N-nitrosoguanidine
2AA= 2-Aminoanthracene