Fatty acids, C18-unsatd., dimers, compds. with coco alkylamines

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

Study period:

1984/1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(Z)-octadec-9-enylamine

EC Number:

204-015-5

EC Name:

(Z)-octadec-9-enylamine

Cas Number:

112-90-3

Molecular formula:

C18H37N

IUPAC Name:

octadec-9-en-1-amine

Details on test material:

- Name of test material (as cited in study report): (Z)-Octa-9-decenylamine
- Test item ID: ODA-FG-11-27-84
- Physical state: cloudy white liquid
- Lot/batch No.: not provided
- Stability under test conditions: responsibility of the sponsor
- Storage condition of test material: room temperature

Method

Target gene:

HGPRT: hypoxanthine-guanine phosphoribosyl transferase

Species / strain

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

- Type and identity of media: Ham's F12 medium, supplemented with 5% FBS, 1% penicillin-streptomycin and 1% L-glutamine
- Properly maintained: yes

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9

Test concentrations with justification for top dose:

First experiment
0.1, 0.5, 1.0, 1.5, 2.0nl/ml without S9
5.0, 6.0, 7.0, 8.0, 9.0 nl/ml with S9

Second Experiment:
1.0, 1.5, 2.0, 2.25, 2.5nl/ml without S9
7.0, 8.0, 9.0, 9.5, 10.0nl/ml with S9

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

ethylmethanesulphonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 5h
- Expression time (cells in growth medium): 7-9days
- Selection time (if incubation with a selection agent): 7 days
- Fixation time (start of exposure up to fixation or harvest of cells): 14 - 16 days

SELECTION AGENT (mutation assays): 6-thioguanine

NUMBER OF REPLICATIONS: 2

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

Evaluation criteria:

The assay is considered positive, if a dose-dependent increase in mutation frequency is observed, with one or more concentrations inducing a mutation frequency, which is at least twice as high as that of the solvent control.

The assay is considered suspect, if no dose response is observed, but one or more doses induce mutant frequencies twice that of the solvent control.

The assay is considered negative, if none of the doses tested induces twice the mutation frequency of the solvent control.

The assay is considered valid, if the cloning efficiency of the solvent controls are no less than 50%. The spontaneous mutation frequency of the solvent control must be within the range of historical control data (0-20 mutants per 10^6 clonable cells). The positive control must induce a mutation frequenc at least three times that of the solvent control.

Results and discussion

Test results

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not affected

RANGE-FINDING/SCREENING STUDIES:
In a range finding study, one culture each of CHO cells was exposed to concenctrations of the test substance from 0.2nl/ml - 10µl/ml without S9, and from 1-30nl/ml with S9. The highest dose was selected to yield 10-30% survival. Of the four lower doses, one was non-toxic.

COMPARISON WITH HISTORICAL CONTROL DATA:
Results from solvent and positive control cultures were within the historical control ranges obtained in 50 CHO/HGPRT mutation assays. No dose response nor reproducible increase in mutant frequency of the treated cells above concurrent solvent controls or historical control data was observed.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Treatment with the test substance led to a very steep dose response curve for cytotoxicity:
Without S9, treatment with 2.0nl/ml gave cloning efficiencies relative to control cultures of 58 - 65% (three independent experiments). Concentrations of 2.25 and 2.5nl/ml were too toxic to clone, while 1.5nl/ml did not induce cell death.
With S9 mix, 9nl/ml gave a relative cloning efficiency of 55 - 71% (three independent experiments). After incubation with 10nl/ml, only 1.5 - 5.5% of the cells survived.

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