7-azatridecane-1,13-diamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 3 FEB 1999 to 5 APR 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

version of 1997

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

7-azatridecane-1,13-diamine

EC Number:

205-593-1

EC Name:

7-azatridecane-1,13-diamine

Cas Number:

143-23-7

Molecular formula:

C12H29N3

IUPAC Name:

bis(6-aminohexyl)amine

Test material form:

solid

Details on test material:

white to slight yellow

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL: the substance appeared to be stable under the conditions of the study, no evidence of instability was observed.

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: TA 97a, TA 98, TA 100, and TA 1535

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Metabolic activation system:

aroclor 1254 induced male rat liver S9-mix

Test concentrations with justification for top dose:

0, 10, 50, 100, 500, 1000, 2500, and 5000 µg/plate (highest concentration is maximum concentration requiered according to guideline)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used:sterile water
- Justification for choice of solvent/vehicle: no evidence for instability was observed

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

9,10-dimethylbenzanthracene

2-nitrofluorene

sodium azide

N-ethyl-N-nitro-N-nitrosoguanidine

other: ICR-191 dihydrochloride (CASRN: 17070-45-0)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Preincubation period: no
- Exposure duration:incubation at 37°C for 48 hours

NUMBER OF REPLICATIONS: 3 plates per concentration

DETERMINATION OF CYTOTOXICITY
- yes

Rationale for test conditions:

AN individual trial must have included a negative and a positive control and at least 5 concentration levels of the test substance for each tester strain and condition.
A data point, concnetration level or trial was excluded from analysis when acceptability criteria were not met - these are as follows:
- all tester strains must have had their strain characteristic specifics and exhibit a characteristic number of revertants per plate in the absence of the test substance
- mean positive control values must exhibit at least a three-fold increase of revertants compared to solvent control
- a minimum of three non-toxic concentrations were requiered (non toxic, i.e. < 50% reduction in mean number of revertants per plate relative to the mean of the concurrent negative control)

Evaluation criteria:

POSITIVE, if the mean number of revertant colonies per plate in at least one strain with or without metabolic activation system was at least two times greater than the mean of concurrent vehicle control and there was a concentration-related increase over the range tested

NEGATIVE, if no two-foldincrease in number of revertant colonies or no concentration-related increase over the range tested

Results not meeting the criteria for positive or negavtive were evaluated using scientific judgment and may have been reported as EQUIVOCAL.

Statistics:

mean number of three plates/concentration level and condition as well as standard deviation was calculated

Results and discussion

Test results

Test results 1

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 2500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 2500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium, other: TA 97a, TA 100, and TA 1535

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 1000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

not examined

Test results 5

Key result

Species / strain:

S. typhimurium, other: TA 97a, TA 98, TA 100, and TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study the test item showed no mutagenic activity in bacterial tester strains.

Executive summary:

In a guideline study according to OECD TG 471 under GLP conditions, mutagenic activity of the test item was investigated in a plate incorporation assay. Salmonella typhimurium strains TA 97a, TA1535, TA98 and TA100 as well as Escherichia coli strain WP2uvrA were exposed to concentrations of 0, 10, 50, 100, 500, 1000, 2500 or 5000 µg test item per plate either with or without metabolic activation system (aroclor induced male rat liver S9 -mix). Three plates at each concentration level were used. No precepitation of the test substance occurred. Cytotoxicity was observed as low as 500 µg/plate in strains tested without metabolic activation system, or as low as 1000 µg/plate in some strains tested with metabolic activation system. All positive controls induced marked increases in frequency of revertant colonies. All negative controls were found to be in an acceptable range. The test item showed no mutagenic activity.