N-[3-(dimethylamino)propyl]docosanamide

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

Study period:

2005-04-05 to 2005-05-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

(Notification No. 77 of JMOL on September 1, 1988 and Notification No. 67 of JMOL on June 2, 1997)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-[3-(dimethylamino)propyl]docosanamide

EC Number:

262-134-8

EC Name:

N-[3-(dimethylamino)propyl]docosanamide

Cas Number:

60270-33-9

Molecular formula:

C27H56N2O

IUPAC Name:

N-[3-(dimethylamino)propyl]docosanamide

Details on test material:

-Name of test material: DIMAPDO
- Physical state: white granular solid
- Analytical purity: 99.1%

Method

Target gene:

Histidine locus (Salmonella typhimurium strains) and tryptophan locus (E. coli strain)

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

other: additional rfa mutation and uvrB mutations in S. typhimurium strains and uvrA mutation in E.coli strain

Metabolic activation:

with and without

Metabolic activation system:

S9Mix

Test concentrations with justification for top dose:

Initial Assay/range-finding: 5, 20, 78, 313, 1250 and 5000 µg/plate
Confirmatory Assay: 39, 78, 156, 313, 625 and 1250 µg/plate for S. typhimurium and 156, 313, 625, 1250, 2500 and 5000 µg/plate for E. coli

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: tetrahydrofuran 100% (Wako Pure Chemical Industries, Ltd., Lot number: CEH5308)
- Justification for choice of solvent/vehicle: tetrahydrofuran was used as the solvent of the test substance by the reason that it could obtain solution, and that the test substance is judged stable. It was confirmed previously that the solvent of tetrahydrofuran did not cause bad influence to growh of the bacterial strains and metabolism of drug-metabolizing enzymes

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

furylfuramide

other: (without metabolic activation)

Controls 2

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

other: 2-aminoanthracene (with metabolic activation)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 hours
- Selection time (if incubation with a selection agent): 48 hours; simultaneous with exposure

SELECTION AGENT (mutation assays): L-histidine (Salmonella tester strains), L-tryptophan (E. coli tester strain)

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: S. typhimurium 1.6-3.0*10E08 cells, E.coli 6.7-7.0*10E08 cells per plate

DETERMINATION OF CYTOTOXICITY
- Method: Toxic effect of the test substance was examined with a stereoscopic microscope

Evaluation criteria:

When the negative control and the positive control values satisfy acceptance criteria and
the test results satisfy acceptance criteria, the test substance was judged positive for mutagenic
activity when clear dose-related increase in the number of the revertant colonies and two-fold
or more increase in the number of the revertant colonies compared with the negative control
were observed with reappearance.

Statistics:

No statistical analysis was used for evaluation of the test results

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

Vehicle 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:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: N/A
- Effects of osmolality: N/A
- Evaporation from medium: N/A
- Water solubility: N/A
- Precipitation: precipitation of the test substance was detected at the dose of 313 µg/plate or more in both the presence and the absence of metabolic activation, but it did not have a bad influence on the observation of the revertant colonies.
- Other confounding effects: the growth of the contaminant was not observed in a result of the sterility test

RANGE-FINDING/SCREENING STUDIES:The preliminary reverse mutation test was performed to determine the most favorable dose levels of the test substance in the reverse mutation test at each of the bacterial strain. The maximum dose of the preliminary reverse mutation test was set at 5000 µg/plate in accordance with the test guideline applied, and total six different dose levels with factor of 4 from the maximum dose were employed.

COMPARISON WITH HISTORICAL CONTROL DATA: The numbers of the revertant colonies of the negative control and the positive control were within the range of the standard value of the historical data in the laboratory

ADDITIONAL INFORMATION ON CYTOTOXICITY:The test substance showed toxic effect in Salmonella typhimurium TA1535 and TA1537
of the absence of metabolic activation at the dose of 625 µg/plate or more. In the test of Salmonella typhimurium TA98 and TA100 in the absence of metabolic activation, similarly, toxic effect of the test substance was observed at the dose of 1250 µg/plate or more. Although, neither dose-related increase in the number of the revertant colonies nor two-fold or more increase in the number of the revertant colonies compared with the negative control was observed in any bacterial strains at any dose levels regardless of the presence or the absence of metabolic activation.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Details on Cytotoxicity:

Toxic effect in S. typhimurium TA1535 and TA1537 in the absence of metabolic activation at 625 µg/plate or more. In the test of S. typhimurium TA98 and TA100 in the absence of metabolic activation toxic effect at a dose of 1250 µg/plate or more.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

The mutagenic activity of DIMAPDO was examined in the reverse mutation test using S. typhimurium tester strains TA98, TA100, TA1535, and TA1537 and E. coli tester strain WP2 uvrA. The test was performed in pre-incubation methods in all bacterial strains in both the presence and the absence of metabolic activation.From the foregoing results, it is concluded that the mutagenic activity of DIMAPDO is considered negative under the test conditions employed.

Executive summary:

In a reverse gene mutation assay in bacteria in a study equivalent to OECD471, strains TA100, TA1535, TA98 and TA1537 of S. Typhimurium were exposed to DIMAPDO in tetrahydrofuran at concentrations of 39, 78, 156, 313, 625 and 1250 µg/plate and Escherichia coli WP2 uvrA were exposed to 156, 313, 625, 1250, 2500 and 5000 µg/plate.

The test was performed with the pre-incubation method in all bacterial strains in both the presence and the absence of metabolic activation.

 

DIMAPDO was tested up to cytotoxic concentrations for S. typhimurium (1250 µg/plate) and to limit concentration (5000 µg/plate) for E.coli. Precipitation of the test substance was detected at a dose of 313 µg/plate or more in both the presence and the absence of metabolic activation, but it did not have a bad influence on the observation of the revertant colonies. The positive controls induced the appropriate responses in the corresponding strains.  There was no evidence of induced mutant colonies over background.

This study is classified as acceptable.