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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

1-(Vinyloxy)octadecane was shown to be non mutagenic in a bacterial reverse mutation assay (Ames test). 1-(Vinyloxy)octadecane was considered to be non mutagenic in the HPRT assay. In the structural chromosome aberrations test in V79 cells (Chinese hamster cell line) in vitro 1 -(vinyloxy)octadecane did not induced chromosomal aberrations.

Link to relevant study records
Reference
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:
1989-01-19 to 1989-03-20
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study. Only four bacterial strains tested (S. typh. TA98, TA100, TA 1535, TA1537). No GLP.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1988
Deviations:
no
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S-9 mix
Test concentrations with justification for top dose:
0; 20; 100, 500; 2500; 5000 µg/plate
Vehicle / solvent:
DMSO
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (2-AA)
Remarks:
with S-9 mix; TA98, TA100,TA1535, TA1537
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
Remarks:
without S-9 mix; TA100, TA1535
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylenediamine
Remarks:
without S-9 mix; TA98
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without S-9 mix; TA1537
Details on test system and experimental conditions:
METHOD OF APPLICATION: Standard plate test
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: 3 plates per dose and control

METHOD OF APPLICATION: Preincubation test
DURATION
- Preincubation period: 20 min at 37°C
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: 3 plates per dose and control

DETERMINATION OF CYTOTOXICITY
- Method: background growth
Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results .
Statistics:
NA
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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
The slight but not dose-dependent increase in the number of his+ revertants using TA 1537 in the 1st experiment could not be confirmed either in a additional standard plate test or in a preincubation assay; therefore the findings of this 1st experiment must be regarded as incidental.
Toxicity: No bacteriotoxic effect (reduced his- background growth) was observed.
Solubility: Complete solubility of test substance in DMSO.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Ames test:

The substance 1-(vinyloxy)octadecane was tested for its mutagenic potential based on the ability to induce back mutations in selected loci of several bacterial strains in the Ames test. The following strains were tested: TA 1535, TA 100, TA 1537, TA 98 with the dose range of 20 µg - 5000 µg/plate (plate incorporation and precincubation test, all strains). The test conditions were as followed: Standard plate test (SPT) and preincubation test (PIT) both with and without metabolic activation (Aroclor induced rat liver S-9 mix).There were no precipitations of the test substance.

The test item showed no bacteriotoxic effect and an increase in the number of his+ or trp+ revertants was not observed both in the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system. According to the results of the present study, 1 -(vinyloxy)octadecane is not mutagenic in the Ames test under the experimental conditions chosen here (BASF, 1989).

HPRT-Test:

No gene mutation assay in mammalian cells is available for 1-(vinyloxy)octadecane. HPRT Tests are available for 3 other Vinylethers, all indicating no mutagenic potential of 1 -(vinyloxy)octadecane.

Hydroxybutylvinylether (CAS 17832 -28 -9, weight of evidence):

The substance Hydroxybutylvinylether was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro. Two independent experiments were carried out, both with and without the addition of liver S9 mix from induced rats (exogenous metabolic activation). According to an initial range-finding cytotoxicity test for the determination of the experimental doses the following doses were tested and evaluated in this study:

1st Experiment: without S9 mix (4-hour exposure period) 0; 150; 300; 600; 1200 μg/mL with S9 mix (4-hour exposure period) 0; 150; 300; 600; 1200 μg/mL

2nd Experiment: without S9 mix (24-hour exposure period) 0; 150; 300; 600; 1200 μg/mL with S9 mix (4-hour exposure period) 0; 450; 600; 900; 1200 μg/mL

After an attachment period of 20-24 hours and a treatment period of 4 hours both with and without metabolic activation and 24 hours without metabolic activation, an expression phase of about 6-8 days and a selection period of about 1 week followed. The colonies of each test group were fixed with methanol, stained with Giemsa and counted. The negative controls gave mutant frequencies within the range expected for the CHO cell line. Both positive control substances, EMS and MCA, led to the expected increase in the frequencies of forward mutations. In this study, after 4 and 24 hours exposure in both experiments in the absence and the presence of metabolic activation no cytotoxicity was observed up to the highest required concentrations tested for gene mutations.

On the basis from the results of the present study, the test substance did not cause any biologically relevant increase in the mutant frequencies both without S9 mix and after adding a metabolizing system in two experiments performed independently of each other.

Thus, under the experimental conditions of this study, Hydroxybutylvinylether is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation (BASF, 2010).

Vinylethylether (CAS 109 -92 -2, weight of evidence):

The study was performed to investigate the potential of vinylethylether to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster. The assay was performed in three independent experiments, using two parallel cultures . The first experiment was performed both, in the presence and absence of metabolic activation . The second experiment was performed in the absence of metabolic activation with a treatment interval of 24 h. The second experiment had to be terminated prematurely due to insufficient cell growth. Therefore, a third experiment was performed as an exact replica of the second experiment .

No visible precipitation of the test article occurred up to the maximal concentration of 720.0 μg/mL (10 mM) . According to the pre-test on toxicity and the molecular weight of the test article the concentration ranges were selected. No relevant toxicity of the test article was observed, neither in the presence nor in the absence of metabolic activation. No substantial or dose dependent increase in mutant colony numbers occurred in both experiments up to the maximal concentration. Appropriate reference mutagens were used as positive controls and showed a distinct increase in induced mutant colonies.

In conclusion, Vinylethylether did not induce gene mutations at the HPRT locus in V79 cells. Therefore, vinylethylether is considered to be non-mutagenic in this HPRT assay (RCC, 1998).

Vinylisobutylether (CAS 109 -53 -5, weight of evidence):

VinyIisobutylether was assessed for its potential to induce gene mutations at the HPRT locus using CHO cells. Two independent experiments were carried out, both with and without exogenous metabolic activation, using identical procedures. Reduced cloning efficiencies were found 18-20 hours post exposure to the test substance at concentrations >= 5 mg/mL both in the absence and in the presence of metabolic activation. No increases in mutant frequency were observed in any of the experiments both in the absence and the presence of metabolic activation.

It is concluded that vinylisobutylether is not mutagenic under the test conditions employed in this in vitro test system (BASF, 1993)

Discussion:

1-(Vinyl)octadecylether was tested for mutagenic activity in a non-GLP bacterial reverse mutation assay according to OECD guideline 471. No signs of mutagenicity were found in this test.

A weight of evidence approach was performed for HPRT tests of three structural analoguous substances: Hydroxybutyvinylether, Vinylethylether and Vinylisobutylether were tested for mutagenic potential in the HPRT mammalian in vitro cell gene mutation assay in Chinese Hamster CHO and V79 cells with and without metabolic activation. None of the three substances was found to show mutagenic potential in the HPRT assay with and without metabolic activation. Therefore, it was concluded that 1-(Vinyl)octadecylether is also non mutagenic in the HPRT assay.

Chromosom Aberration Test

The test item 1-(vinyloxy)octadecane, dissolved in THF, was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in two independent experiments. The following study design was performed without S9 mix and with S9 mix. In each experimental group two parallel cultures were set up. 100 metaphases per culture were evaluated for structural chromosome aberrations, except for the positive control in Experiment II at preparation interval 28 hours without S9 mix, where 50 metaphases were evaluated.

The highest applied concentration (3707.0 µg/mL; approx.10 mM) was chosen with regard to the molecular weight and the purity (80 %) of the test item and with respect to the current OECD Guideline 473. Dose selection for the cytogenetic experiments was performed considering the toxicity data. No cytotoxicity was observed up to the highest applied concentration. No clastogenicity was observed at the concentrations evaluated either with or without metabolic activation. No evidence of an increase in polyploid metaphases was found after treatment with the test item as compared to the frequencies of the control cultures.

Appropriate mutagens (EMS and CPA) were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations.

In conclusion, the test item did not induce structural chromosome aberrations in V79 cells (Chinese hamster cell line) in vitro.

Therefore, 1-(vinyloxy)octadecane is considered to be non-clastogenic in this chromosome aberration test in the absence and presence of metabolic activation, when tested up to the highest required concentration.

Justification for classification or non-classification

Classification is not warranted according to the criteria of EU Classification, Labelling and Packaging of Substances and Mixtures (CLP, GHS) Regulation (EC) No 1272/2008.