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Toxicological information

Genetic toxicity: in vitro

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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:
05 May - 26 Sep 1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study performed according to acceptable standards including GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report date:
1989

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
yes
Remarks:
- limited documentation
GLP compliance:
yes
Type of assay:
bacterial forward mutation assay

Test material

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
IUPAC Name:
octadec-9-en-1-amine
Constituent 2
Reference substance name:
(Z)-octadec-9-en-1-amine
IUPAC Name:
(Z)-octadec-9-en-1-amine
Constituent 3
Reference substance name:
Oleyl Alkylamines
IUPAC Name:
Oleyl Alkylamines
Details on test material:
- Name of test material (as cited in study report): Oleylamine

Method

Target gene:
thymidine kinase locus
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Aroclor induced rat liver S9-mix
Test concentrations with justification for top dose:
initial tests:
without metabolic activation: 0.13-10 nL/mL
with metabolic activiation: 1.3-100 nL/mL

confirmatory test.
without metabolic activation: 0.2-2 nL/mL
with metabolic activation: 1.5-15 nL/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
aceton
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: ethylmethanesulphonate in DMSO (nonactivated), 7,12-dimethylbenzanthracene in DMSO (activated)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in suspension

DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 2 days

SELECTION AGENT (mutation assays): trifluorthymidine (final concentration: 4 µg/mL)

NUMBER OF REPLICATIONS: two initial assays (using single cultures per dose) were performed to achieve the desired range of toxicity and one confirmatory assay (using duplicate cultures per dose) was carried out

DETERMINATION OF CYTOTOXICITY
- Method: Toxicity was determined by comparing the cell population growth at each dose level with that of the solvent controls.

Evaluation criteria:
Positive - if there is a positive dose response and one or more of the 3 highest doses in the 10% or greater Total Growth range exhibit a mutant frequency wich is two-fold greater tahn the background level. All data including that from cultures with less than 10% Total Growth will be used to establish the dose response relationship. The first assay and the confirmatory assay must both demonstrate a positive response to call a test article a positive mutagen.

The spontaneous mutant frequency of the solvent control must be between 0.2 and 1.0 per 10000 surviving animals.
The plating efficiency of the solvent controls must be greater than 50%.
Mutant frequency of the positive control at least twice that of the solvent control.

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
100% toxicity at 100 nL/mL (with metabolic activation); 100% toxicity at 10 nL/mL (without metabolic activation)
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
Confirmatory assay:
None of the nonactivated cultures that was cloned exhibited a mutant frequency which was two times the mean mutant frequency of the solvent controls. The total growths of these cultures ranged from 6% to 109%. A dose dependent response was not noted in the treated cultures. None of the S9 activated cultures that was cloned showed a mutant frequency which was twice the mean mutant frequency of the solvent controls. The total growth of these cultures ranged from 2% to 115%. A dose dependent response was not noted in the treated cultures.
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

Based on the study results the test article oleylamine was negative in both the presence and absence of exogenous metabolic activation in this Mouse Lymphoma Mutagenesis Assay.
Executive summary:

The test article Oleylamine was tested in the L5178Y TK+/- Mouse Lymphoma Mutagenesis Assay in the presence and absence of Aroclor induced rat liver S-9. Two initial and one confirmatory assays were conducted.

The nonactivated cultures selected for cloning in the first initial assay were treated with doses of 0.32 to 0.13 nl/ml and exhibited total growths from 75% to 99%. The S-9 activated cultures selected for cloning in the first initial assay were treated with doses of 10 to 1.3 nl/ml which produced from 6% to 106% total growths. None of the nonactivated or S-9 activated cultures that was cloned exhibited a mutant frequency which was at least twice the mean mutant frequency of the solvent controls. A dose dependent response was not noted in the treated cultures.

The nonactivated cultures selected for cloning in the second initial assay were treated with doses of 1.8 to 0.13 nl/ml and exhibited total growths from 3% to 110%. The S-9 activated cultures selected for cloning in the second initial assay were treated with doses of 13 to 1.3 nl/ml which produced from 13% to 107% total growths. None of the nonactivated or S-9 activated cultures that was cloned exhibited a mutant frequency which was at least twice the mean mutant frequency of the solvent controls. A dose dependent response was not noted in the treated cultures.

The nonactivated cultures selected for cloning in the confirmatory assay (duplicate cultures) were treated with doses of 1.0 to 0.2 nl/ml and exhibited total growths from 6% to 109%. The S-9 activated cultures selected for cloning in the confirmatory assay were treated with doses of 11 to 1.5 nl/ml which produced from 2% to 115% total growths. None of the nonactivated or S-9 activated cultures that was cloned exhibited a mutant frequency which was at least twice the mean mutant frequency of the solvent controls. A dose dependent response was not noted in the treated cultures.

The results indicate that, under the conditions of these mutagenicity tests, the test article Oleylamine was negative in both the presence and absence of exogenous metabolic activation in the Mouse Lymphoma Mutagenicity Assay.

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