N,N,N',N'-tetramethyltrimethylenediamine

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: Guideline study, GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(only 2-Aminoanthracene as positive control with S9-mix)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N,N',N'-tetramethyltrimethylenediamine

EC Number:

203-818-8

EC Name:

N,N,N',N'-tetramethyltrimethylenediamine

Cas Number:

110-95-2

Molecular formula:

C7H18N2

IUPAC Name:

N,N,N',N'-tetramethyltrimethylenediamine

Details on test material:

- Name of test material (as cited in study report): N,N,N',N'-Tetramethyl-1,3-propandiamin
- Physical state: colourless liquid
- Analytical purity: 99.5%
- Lot/batch No.: Tank 90
- Storage condition of test material: Room temperature (N2 conditions)
- Substance number: 96/648

Method

Target gene:

His- 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:

The S9 fractions were prepared from the liver of Aroclor 1254-induced male Sprague-Dawley rats.

Test concentrations with justification for top dose:

First experiment (standard plate test): 0, 20, 100, 500, 2500, 5000 µg/plate.
Second experiment (preincubation test): 0, 4, 20, 100, 500, 2500 µg/plate (Salmonella typhimurium); 0, 20, 100, 500, 2500, 5000 µg/plate (E. coli).

Vehicle / solvent:

- Vehicle/solvent used: water

Controls

Untreated negative controls:

yes

Remarks:

Sterility control

Negative solvent / vehicle controls:

yes

Remarks:

Vehicle control

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S-9mix: all strains 2-aminoanthracene; without S-9 mix: strains TA100, TA1535: N-methyl-N'-nitro-N-nitrosoguanidine; TA98: 4-nitro-o-phenylendiamine; TA1537: 9-aminoacridine; E.coli WP2 uvrA: N-ethyl-N'-nitro-N-nitrosoguanidine.

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h
NUMBER OF REPLICATIONS: 3 per dose

METHOD OF APPLICATION: standard plate test
DURATION
- Exposure duration: 48 - 72 h
NUMBER OF REPLICATIONS: 3

Cytotoxicity is evaluated according to a decrease in the number of revertants, clearing or diminution of the background lawn and/or reduction in the titer.

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 S9-mix or after adding a metabolizing system.
A test substance is generally considered non-mutagenic in this test if: The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

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:

cytotoxicity

Remarks:

A weak bacteriotoxic effect was occasionally observed depending on the strain and test conditions from about 2500 µg/plate onward

Vehicle controls validity:

valid

Untreated negative 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:

cytotoxicity

Remarks:

A weak bacteriotoxic effect was occasionally observed depending on the strain and test conditions from about 2500 µg/plate onward.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No substance precipitation was found.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test:

Dose (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli WP2 uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	20±2	20±1	153±17	158±9	13±4	12±3	31±3	45±3	37±7	36±5
20	18±3	19±4	156±9	162±3	8±3	12±2	33±3	42±4	34±5	42±7
100	18±2	19±1	133±7	163±6	10±2	12±3	31±4	43±4	37±7	41±9
500	17±2	19±3	151±6	154±16	4±1	11±1	29±1	36±2	38±8	44±4
2500	18±1	14±3	160±17	161±17	7±4	9±2	27±2	32±4	39±7	39±7
5000	11±1	10±2	145±10	139±23	9±2	9±2	11±3	24±5	30±3	31±1
2 -AA	-	149±7	-	1747±62	-	179±2	-	1237±32	-	317±37
MNNG	955±36	-	858±42	-	-	-	-	-	-	-
AAC	-	-	-	-	750±6	-	-	-	-	-
NOPD	-	-	-	-	-	-	1104±77	-	-	-
ENNG	-	-	-	-	-	-	-	-	1185±189	-

Mean ± SD

 

Preincubation-test:

Dose (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli WP2 uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	20±1	19±2	110±6	105±1	11±1	10±1	24±3	38±5	23±2	33±2
4	20±2	17±1	115±15	125±3	10±2	11±2	29±2	36±4	23±6	32±3
20	20±1	16±1	122±18	120±10	10±1	10±1	28±2	31±3	23±1	26±2
100	17±1	17±1	116±5	111±4	8±1	10±1	25±2	28±3	24±3	27±5
500	19±1	19±1	90±8	104±10	10±1	10±1	24±2	27±4	24±3	21±3
2500	11±3	15±1	23±3	100±7	4±3	9±1	3±1	30±3	14±3	21±4
2-AA	-	200±4	-	1164±54	-	99±3	-	1015±15	-	170±8
MNNG	1284±34	-	1157±54	-	-	-	-	-	-	-
AAC	-	-	-	-	701±26	-	-	-	-	-
NOPD	-	-	-	-	-	-	992±5	-	-	-
ENNG	-	-	-	-	-	-	-	-	896±13	-

Mean ± SD

2-AA: 2-aminoanthracene; MNNG; N-methyl-N-nitro-N-nitrosoguanidine; ENNG; N-ethyl-N-nitro-N-nitrosoguanidine; NOPD: 4-nitro-o-phenylendiamine; AAC: 9-aminoacridine chloride monohydrate

Under the conditions tested the test substance is non mutagenic in the bacterial AMES-test.

The positive control gave the expected response.

 

Applicant's summary and conclusion

Executive summary:

The potential of N,N,N',N'‑tetramethyl‑1,3‑propanediamine (TMPDA) to induce reverse mutation in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and Escherichia coli WP2 uvrA was evaluated in accordance with the OECD guidelines no. 471 and 472. TMPDA did not induce any noteworthy increase in the number of revertants, both with and without S9 mix, in any strains. Under these experimental conditions, TMPDA did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium and Escherichia coli.