Ethyl (2E,4Z)-2,4-decadienoate

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:

October 20-31, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl (2E,4Z)-2,4-decadienoate

EC Number:

221-178-8

EC Name:

Ethyl (2E,4Z)-2,4-decadienoate

Cas Number:

3025-30-7

Molecular formula:

C12H20O2

IUPAC Name:

ethyl deca-2,4-dienoate

Test material form:

not specified

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0007631647
- Expiration date of the lot/batch: 10/2018
- Purity test date: September 27, 2016


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, protected from light, then under nitrogen
- Solubility and stability of the test substance in the solvent/vehicle: soluble in DMSO

Method

Target gene:

S. typhimurium: histidine dependence
E. coli: tryptophan locus

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

Test concentrations with justification for top dose:

5.00, 15.0, 50.0, 150, 500, 1500, and 5000 ug/plate
No precipitate was observed, some toxicity was noted at 500 ug/plate and above.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility of the test substance and compatibility with target cells

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)
- Cell density at seeding (if applicable): 2.0 - 5.2 x 10^9 cells per milliliter
- Fixation time (start of exposure up to fixation or harvest of cells): 12 hrs

DURATION
- Exposure duration: 48-72 hrs

SELECTION AGENT (mutation assays): BBL selective agar

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: all

DETERMINATION OF CYTOTOXICITY
- Method:relative growth
- Any supplementary information relevant to cytotoxicity: A dose level was considered toxic if there was at least a 50% reduction in mean number of revertants per plate and a dose-dependent drop in the revertant count, and moderate to significant reduction in the background lawn.

Rationale for test conditions:

In the initial assay, dose levels of 1.50, 5.00, 15.0, 50.0, 150, 500, 1500, and 5000 ug/plate were evaluated. Toxicity was observed beginning at 500 ug/plate. No precipitation was observed, thus the maximum dose for the confirmatory assay was 5000 ug/plate.

Evaluation criteria:

All S. typhimurium strains must demonstrate presence of rfa mutations and deletion in the uvrB gene. S. typhimuirum strains TA98 and TA100 must demonstrate presence of the pKM101 plasmid R-factor. E. coli culture must demonstrate deletion in the uvrA gene.

Mean number of spontaneous revertants: 10-50 (TA98), 80-240 (TA100), 5-45 (TA1535), 3-21 (TA1537), 10-60 (WP2 uvrA)

Tester strain culture titers must be >= 0.3 x 10^9 cells/mL.

Postitive controls must exhibit at a 3-fold increase in the number of revertants.

A minimum of 4 non-toxic dose levels.

Statistics:

Mean, standard deviation

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxicity was observed starting at 500 ug/plate.

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: None

RANGE-FINDING/SCREENING STUDIES: In the initial assay, dose levels of 1.50, 5.00, 15.0, 50.0, 150, 500, 1500, and 5000 ug/plate were evaluated. Toxicity was observed beginning at 500 ug/plate. No precipitation was observed.


HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: see table below
- Negative (solvent/vehicle) historical control data: see table below

Any other information on results incl. tables

Historical Control Data (2015 Revertants per Plate)

Strain	TA98	TA98	TA100	TA100	TA1535	TA1535	TA1537	TA1537	WP2 uvrA	WP2 uvrA
Control	Negative	Positive	Negative	Positive	Negative	Positive	Negative	Positive	Negative	Positive
Mean (no S9)	16	190	90	697	13	624	7	392	25	336
Standard Deviation (no S9)	5	191	12	172	5	196	3	292	8	112
Minimum (no S9)	6	42	62	239	2	50	1	24	7	89
Maximum (no S9)	43	2468	233	1767	35	2509	20	2887	73	1026
95% Confidence Limit (no S9)	6 -26		66 -114		3 -23		1 -13		9 -41
Mean (S9)	23	329	98	671	13	137	9	73	28	352
Standard Deviation (S9)	7	176	15	284	5	110	3	53	8	117
Minimum (S9)	5	51	63	138	3	24	2	19	10	78
Maximum (S9)	53	1786	15 7	2692	33	1060	23	574	96	1409
95% Confidence Limit (S9)	9 -37		68 -128		3 -23		3 -15		12 -44

Applicant's summary and conclusion

Conclusions:

Not mutagenic either in the presence or absence of S9.

Executive summary:

The mutagenicity of ethyl decadienoate was tested in an OECD Guideline 471 Ames test. The mutagenicity was tested both in the presence and absence of S9 using DMSO as a solvent. Doses of 5.00, 15.0, 50.0, 150, 500, 1500, and 5000 ug/plate were tested. Toxicity was seen at doses of 500 ug/plate or higher. The test substance was not mutagenic either in the presence or absence of S9.