Reaction mass of N,N-dimethyldodecanamide and N,N-dimethyltetradecanamide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to guideline, well conducted (GLP) and documented

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

67359-57-3

Cas Number:

67359-57-3

IUPAC Name:

67359-57-3

Constituent 2

Reference substance name:

N,N-Dimethyldecan-1-amide, mixture with N,N-Dimethyloctanc-1-amide

IUPAC Name:

N,N-Dimethyldecan-1-amide, mixture with N,N-Dimethyloctanc-1-amide

Details on test material:

- Chemical name: mixture of N,N-Dimethydecan-1-amide and N,N-Dimethyloctan-1-amide
- Physical state: liquid
- Storage condition of test material: roomtemperature

Method

Target gene:

HGPRT locus

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

V79 cell line was originally derived from the lung of a
male Chinese hamster (Chu and Mailing, 1968) kind gift from Prof. G. Speit, University of Ulm, Germany

Metabolic activation:

with and without

Metabolic activation system:

S9 Mix Aroclor 1254 induced from Wistar rats

Test concentrations with justification for top dose:

Cytotoxicity test: 7.9, 15.7, 31.3, 62.5, 125.0, 250.0, 500.0, 750.0, 1000.0 µg/ml
Main test: 25.0, 50.0, 100.0, 125.0, 150.0, 200.0, 250.0µg/ml

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: solubility

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: Ethylmethanesulfonate (EMS) and Dimethylbenzanthracene (DMBA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium (containing 2% FCS)

according to Guideline

DURATION
- Preincubation period: 16-24h
- Exposure duration: 5h
- expression period: 6 days
- Selection time (if incubation with a selection agent): 7 days

SELECTION AGENT (mutation assays): 6-Thioguanine
STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: triplicates for cytotoxicity
Main test (repeated completly two times ): 8 dishes (from 1 flask) were examined, for each dosage 2 flask were used.

NUMBER OF CELLS EVALUATED:
for each dose level one flask with 4*10e6 cells were avaiable.

DETERMINATION OF CYTOTOXICITY
- Method: cell count, average count, cytotoxicity was expressed by comparison of
colonies in treated cultures versus vehicle control cultures (relative cloning efficiency)

Evaluation criteria:

The number of 6-TG resistant colonies in the mutation assay dishes and the number of colonies in the
cloning efficiency dishes were determined.Colonies with 50 cells or less were excluded

Statistics:

Weighted analysis of variance as well as to a weighted recursive regression, both with Poisson derived weights. Weighted analysis of
variance followed by pairwise comparisons using Dunnetts test.

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

up to cytotoxic concentration

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Precipitation of the test article after addition of the test article-Ethanol solution to
the medium starting to be evident at 1500 µg/ml. Therefore, a preliminary cytotoxicity test was conducted with a series of
9 concentrations ranging from 7.9 µg/ml to 1000 µg/ml

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

the test article is considered to be nonmutagenic in the V79-HGPRT Forward Mutation Assay both with and without metabolic activation.

Executive summary:

The test material, confidential substance name, was assayed for mutagenic activity at the HGPRT locus in V79 cells from 25 to 250 µg/ml both, with and without metabolic activation according to OECD guideline 476. Under both treatment conditions, cytotoxic effects were induced . The vehicle control mutant frequencies were all in the normal range of background frequencies for the assay. In contrast, the positive controls EMS and DMBA induced a distinct mutagenic effect in mutant frequency, which was significantly increased over the negative controls demonstrating the sensitivity of the test system and the ability to detect known mutagens.