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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Data published in 1987
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
significant methodological deficiencies
Remarks:
Strain TA102, WP2 uvrA or WP2 uvrA pKM101 not tested

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity tests: III. Results from the testing of 255 chemicals
Author:
Zeiger E, Anderson B, Haworth S, Lawlor T, Mortelmans K & Speck W
Year:
1987
Bibliographic source:
Environmental Mutagenesis Volume 9, Supplement 9:1-110

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1989
Deviations:
yes
Remarks:
Salmonella strain TA102 or E.coli strains WP2 uvrA or WP2 uvrA pKM101 not tested
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N,N',N'-tetramethylethylenediamine
EC Number:
203-744-6
EC Name:
N,N,N',N'-tetramethylethylenediamine
Cas Number:
110-18-9
Molecular formula:
C6H16N2
IUPAC Name:
[2-(dimethylamino)ethyl]dimethylamine
Test material form:
liquid
Details on test material:
clear, colourless to pale yellow
Specific details on test material used for the study:
CAS number: 110-18-9
Supplier: Sigma
Purity: 99%

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S-9 (Sprague Dawley and Syrian Hamster)
Test concentrations with justification for top dose:
10000 ug/plate. The highest concentration was tested above the recommended maximum test concentration of 5000 ug/plate; toxicity was apparent in some strains.
Vehicle / solvent:
Water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine (strain TA98 in the absence of S-9) and 2-aminoanthracene (all strains in the presence of S-9)
Details on test system and experimental conditions:
Bacterial strains were received from Dr. Ames (University of California). Bacterial cultures were grown overnight at 37 degrees C, with shaking, in Oxoid broth. Phenotypes were analysed. The test material, bacteria and S-9 mix or buffer were incubated at 37 degrees C, with shaking, for 20 minutes. The top agar was added and the resulting contents mixed and poured on to Vogel Bonner test plates. The revertant colonies were counted following 2 days incubation at 37 degrees C. Counting was automatic, or hand-counted if the presence of precipitate.
Evaluation criteria:
Toxicity was considered to be evidence of a decrease in revertant colonies or a diminution of the backgound bacterial lawn. A positive mutagenic response was considered to be a dose related increase in the number of revertant colonies over the concurrent control. Scientific judgement was applied in interpreting increases in revertant numbers at a single concentration or a non-concentration related increase. Reprodubility between two experimental occassions were also considered.
Statistics:
None

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
in the presence of hamster S-9
Vehicle controls validity:
not specified
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
in the absence of S-9 and in the presence of hamster S-9
Vehicle controls validity:
not specified
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
not specified
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
not specified
Untreated negative controls validity:
not applicable
Positive controls validity:
valid

Any other information on results incl. tables

Revertant numbers in the absence of metabolic activation

 

Strain

Concentration (mcg/plate)

TA98

TA100

TA1535

TA1537

0 (water)

16

98

15

6

100

11

98

17

4

333

19

78

16

7

1000

11

89

17

7

3333

13

78

15

6

10000

13

78 S

12

5

Positive control

637

360

340

298

 

Revertant numbers in the presence of 10% metabolic activation (hamster liver)

 

Strain

Concentration (mcg/plate)

TA98

TA100

TA1535

TA1537

0 (water)

25

117

7

8

100

31

114

7

5

333

37

115

10

3

1000

28

107

7

7

3333

27

127

15

6

10000

0 S

81 S

17

5

Positive control

579

1207

460

341

 

Revertant numbers in the presence of 10% metabolic activation (rat liver)

 

Strain

Concentration (mcg/plate)

TA98

TA100

TA1535

TA1537

0 (water)

24

115

8

5

100

28

107

12

10

333

22

115

9

4

1000

25

109

7

5

3333

17

106

12

4

10000

22

107

6

5

Positive control

131

449

200

129

Applicant's summary and conclusion

Conclusions:
It was concluded that the test material was not mutagenic under the conditions of testing.
Executive summary:

An Ames testr was conducted using pre-incubation methodology. N,N,N'N'-Tetramethylethylenediamine was tested at concentrations of 100, 333, 1000, 3333 and 10000 mcg/plate in the absence of metabolic activation, in the presence of 10% Aroclor-1254 induced rat liver S-9 and in the presence of 10% Aroclor-1254 induced hamster liver S-9. Testing was conducted in strains TA98, TA100, TA1535 and TA1537. There was some evidence of toxicity at the highest concentration tested in some of the tester strains. There was no evidence of increases in the number of revertants for any of the strains in either the absence or in the presence of metabolic activation (rat or hamster).