3-cyclohexylaminopropylamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Ames test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1977-12-02 to 1978-01-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Sample A-5297 for strain TA-1535 was retested at varous concentrations (see table) in May 1978

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-cyclohexylaminopropylamine

EC Number:

222-001-7

EC Name:

3-cyclohexylaminopropylamine

Cas Number:

3312-60-5

Molecular formula:

C9H20N2

IUPAC Name:

N1-cyclohexylpropane-1,3-diamine

Test material form:

liquid

Details on test material:

3-Cyclohexylaminopropylamine from Performance Chemicals, lot number: 68-219-BC

Method

Species / strain

Species / strain / cell type:

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

Remarks:

Salmonella typhimurium TA-1538 and Saccharomyces cerevisiae: D4 was also tested

Metabolic activation:

with and without

Metabolic activation system:

Reaction mixture containing TPN, G6P, Sodium phosphate, MgCl2, KCL and homogenate fraction
S9 Homogenate

Test concentrations with justification for top dose:

The compound was tested over a series of concentrations such that there was either quantitative or qualitative evidence of some chemically-induced physiological effects at the high dose level. The dose range employed for the evaluation of this compound was from 0.001 µl to 5 µl per plate.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

2-nitrofluorene

other: Methylnitrosoguanidine (MNNG) in Water or Saline, non-activated; Quinacrine mustard (QM) in Water or Saline, non-activated; 2-Anthramine (ANTH) in DMSO, activated; 8-Aminoquinoline (AMQ) in DMSO, activated

Details on test system and experimental conditions:

-Plate- Test (Overlay Method)
Approximately 10^8 cells from an overnight culture of each indicator strain were added to separate test tubes containing 2.0 mL of molten agar supplemented with biotin and a trace of histidine. For nonactivation tests, at least four dose levels of the test compound were added to the contents of the appropriate tubes and poured over the surfaces of selective agar plates. In activation tests, a minimum of four different concentrations of the test chemical were added to the appropriate tubes with cells. Just prior to pouring, an aliquot of reaction mixture (0.5 mL containing the 9,000 x g liver homogenate) was added to each of the activation overlay tubes, which were then mixed, and the contents poured over the surface of a minimal agar plate and allowed to solidify. The plates were incubated for 48 hours at 37C, and scored for the number of colonies growing on each plate. The concentrations of all chemicals are given in the Results Section. Positive and solvent controls using both directly active positive chemicals and those that require metabolic activation were run with each assay.

- Recording -and Presenting -Data
The numbers of colonies on each plate were counted and recorded on printed forms. These raw data were analyzed in a computer program and reported on a printout. The results are presented
as revertants per plate for each indicator strain employed in the assay. The positive and the solvent controls are provided as reference points. Other relevant data are provided on the computer printout

Evaluation criteria:

Because the procedures used to evaluate the mutagenicity of the test chemical are semi quantitative, the criteria used to determine positive effects are inherently subjective and are based primarily on a historical data base. Most data sets are evaluated using the following criteria:
Strains TA-1535, TA-1537, -and TA-1538
If the solvent control value is within the normal range, a chemical that produces a positive dose response over three
concentrations with the lowest increase equal to twice the solvent control value is considered to be mutagenic.
Strains -TA-98, -TA-100, -and- D4
If the solvent control value i s within the normal range, a chemical that produces a positive dose response over three
concentrations with the highest increase equal to twice the solvent control value for TA-100 and two to three times the
solvent control value for strains TA-98 and 04 is considered to be mutagenic. For these strains , the dose response
increase should start at approximately the solvent control value.
Pattern
Because TA-1535 and TA-100 were both derived from the same parental strain (G-46) and because TA-1538 and TA-98 were
both derived from the same parental strain (D3052), there is a built-in redundancy in the microbial assay. In general
the two strains of a set respond to the same mutagen and such a pattern is sought. It is also anticipated that if a
nonactivation tests it will generally do so in activation tests. While similar response patterns are not required for all mutagens, they can be used to enhance the reliability of an evaluation decision.
Reproducibility
If a chemical produces a response in a single test that cannot be reproduced in one or more additional runs, the
initial positive test data loses significance. The preceding criteria are not absolute and other extenuating
factors may enter into a final evaluation decision.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not examined

True negative controls validity:

not specified

Positive controls validity:

not specified

Test results 3

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

not specified

Test results 4

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

not specified

Test results 5

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

not specified

Test results 6

Key result

Species / strain:

Saccharomyces cerevisiae

Remarks:

D4

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

The data for strain TA-1535 was judged to be equivocal and therefore was re-tested at Litton Bionetics. The results were clearly negative for genotoxicity (see table).

Any other information on results incl. tables

Results of the re-test of strain TA-1535

Concentration	Revertants per plate, nonactivation test	Revertants per plate, activation test
Solvent control	26	30
Positive control	940	285
0.001	25	27
0.01	33	35
0.1	31	35
1.0	35	29
5.0	36	35
10.0	0	9
20.0	0	0

Applicant's summary and conclusion

Conclusions:

The test compound did not demonstrate mutagenic activity in any of the assays conducted in the evaluation and was considered as not mutagenic under the test conditions.

Executive summary:

The  compound was tested in an Ames test in six different strains with and without activation incl. positive and negative control. in summary, the test compound did not demonstrate mutagenic activity in any of the assays conducted in the evaluation and was considered as not mutagenic under the test conditions.