2-Propenoic acid, heptadecyl ester, branched

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Reference 2

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

heptadecyl (branched) acrylate

EC Number:

810-816-6

Cas Number:

1473386-36-5

Molecular formula:

C20H38O2

IUPAC Name:

heptadecyl (branched) acrylate

Constituent 2

Reference substance name:

C17 Acrylate

IUPAC Name:

C17 Acrylate

Details on test material:

- Name of test material (as cited in study report): C17 Acrylate
- Physical state: liquid, colorless, clear
- Analytical purity: 96.2 %
- Lot/batch No.: S731910108

Method

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9 mix, prepared for rat livers

Test concentrations with justification for top dose:

3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine; aminoanthracene

Details on test system and experimental conditions:

For each strain and dose level, including the controls, three plates were used.
The following materials were mixed in a test tube and poured onto the selective agar plates:
Experiment I (Plate Incorporation)
100 μL Test solution at each dose level (solvent or reference mutagen solution (positive control)),
500 μL S9 mix (for test with metabolic activation) or S9 mix substitution buffer (for test without metabolic activation),
100 μL Bacteria suspension (cf. test system, pre-culture of the strains),
2000 μL Overlay agar
Experiment II (Pre-Incubation)
In the pre-incubation assay 50 μL test solution or solvent or 100 μL reference mutagen solution (positive control)), 500 μL S9 mix / S9 mix substitution buffer and 100 μL bacterial suspension were mixed in a test tube and incubated at 37 °C for 60 minutes. After pre-incubation 2.0 mL
overlay agar (45 °C) was added to each tube. The mixture was poured on minimal agar plates.
After solidification the plates were incubated upside down for at least 48 hours at 37 °C in the dark. In parallel to each test a sterile control of the test item was performed and documented in the raw data. Therefore, 100 μL (experiment I) and 50 μL (experiment II) of the stock solution, 500 μl
S9 mix / S9 mix substitution buffer were mixed with 2.0 mL overlay agar and poured on minimal agar plates.

Evaluation criteria:

A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.

Statistics:

A statistical analysis of the data is not mandatory.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Additional information on results:

Precipitation of the test item in the overlay agar on the incubated agar plates was observed from 1000 to 5000 μg/plate in experiment I with and without S9 mix and in experiment II without S9 mix. In experiment II with S9 mix precipitation was observed from 2500 to 5000 μg/plate. The undissolved particles had no influence on the data recording.
The plates incubated with the test item showed normal background growth up to 5000 μg/plate with and without S9 mix in all strains used. Only in experiment I reduced background growth was observed in strain TA 1537 from 1000 to 5000 μg/plates in the absence of metabolic activation.
No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in the test groups with and without metabolic activation in both experiments.
Only in experiment I a reduction in the number of revertants (below the indication factor of 0.5), was observed in strain TA 1537 from 1000 to 5000 μg/plates in the absence of metabolic activation. No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with C 17 Acrylate at any dose level, neither in the presence nor absence of metabolic activation (S9 mix).

Applicant's summary and conclusion

Conclusions:

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Therefore, C 17 Acrylate is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.