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Toxicological information

Genetic toxicity: in vitro

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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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(only 2-Aminoanthracene as positive control with S9-mix)
Qualifier:
according to
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

Reference
Name:
Unnamed
Type:
Constituent
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 / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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 resultsopen allclose all
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
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.