Reactionmass of dodecan-2-yl prop-2-enoate and dodecan-3-yl prop-2-enoate and dodecan-4-yl prop-2-enoate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

July, 1997

Deviations:

no

Remarks:

No deviations occurred that had a negative impact on integrity of the study.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reactionmass of dodecan-2-yl prop-2-enoate and dodecan-3-yl prop-2-enoate and dodecan-4-yl prop-2-enoate

EC Number:

947-818-2

Cas Number:

1612783-92-2

Molecular formula:

C15H28O2

IUPAC Name:

Reactionmass of dodecan-2-yl prop-2-enoate and dodecan-3-yl prop-2-enoate and dodecan-4-yl prop-2-enoate

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M Company, Lot 6
- Expiration date of the lot/batch: 04 October, 2019
- Purity test date: 04 October, 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temp
- Stability under test conditions: No data
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was prepared in 200 proof ethanol at a concentration of 50 mg/mL on the day of each assay. The lower concentrations were prepared by serial dilution with the vehicle to attain the appropriate concentration for testing.

FORM AS APPLIED IN THE TEST: Prepared in ethanol

Method

Target gene:

Histidine and tryptophan operon

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9

Test concentrations with justification for top dose:

0 (vehicle), 50, 100, 250, 500, 1000, 2500, and 5000 μg/plate. Precipitates were observed at 2500 ug/plate and higher.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: Test article solubility and tester strain compatibility.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

200 proof ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

other: See remarks

Details on test system and experimental conditions:

METHOD OF APPLICATION:in agar (plate incorporation)
- Cell density at seeding (if applicable):

DURATION
- Preincubation period: Overnight culture of the tester strains
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2.5 days
- Selection time (if incubation with a selection agent): 2 days
- Fixation time (start of exposure up to fixation or harvest of cells): 2 days

SELECTION AGENT (mutation assays): Histidine and tryptophan-minimal agar.

NUMBER OF CELLS EVALUATED: All colonies on all plates were counted.

DETERMINATION OF CYTOTOXICITY
- Method: background lawn health

Rationale for test conditions:

Per OECD 471.

Evaluation criteria:

The test substance was considered positive for mutagenicity if it induced an increase of revertants per plate with increasing concentration. The increases should be at least 2 times the vehicle control background frequency for strains with high spontaneous levels (i.e., TA100) and 3 times for those with low spontaneous levels (TA1537, TA98, TA1535, and WP2 uvrA). These increases should be seen in at least 2 or more successive concentrations or the response should be repeatable at a single concentration.

The test substance was considered to be negative for inducing mutagenicity if it did not induce a response which fulfills the criteria for a positive response.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: None observed
- Effects of osmolality: None observed
- Precipitation: Observed at concentrations greater than 2500 ug/plate

Applicant's summary and conclusion

Conclusions:

 No increase in revertant colonies was observed in any strain in the presence or absence of metabolic activation. MTDID 44430 was not mutagenic under the conditions of this assay.

Executive summary:

The mutagenicity potential of MTDID 44430 was evaluated in the bacterial reverse mutation assay (Ames assay).  The study was conducted according to OECD 471 (1997) in compliance with OECD GLP. S. typhimurium strains TA98, TA100, TA1535, and TA1537 and E.coli strain WP2 uvrA were utilized in the presence and absence of metabolic activation (Aroclor 1254 -induced rat liver S9 mix). A stock solution of MTDID 44430 was prepared in 200 proof ethanol and administered at concentrations up to 5000 ug/plate. The vehicle control for all assays was 200 proof ethanol. Precipitation was observed at concentrations of 2500 ug/plate and greater.  No increase in revertant colonies was observed in any strain in the presence or absence of metabolic activation. MTDID 44430 was not mutagenic under the conditions of this assay.