Propyl oleate

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:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Propyl (9Z,12Z)-octadeca-9,12-dienoate

Molecular formula:

C21H38O2

IUPAC Name:

Propyl (9Z,12Z)-octadeca-9,12-dienoate

Constituent 2

Reference substance name:

Propyl oleate

EC Number:

203-885-3

EC Name:

Propyl oleate

Cas Number:

111-59-1

Molecular formula:

C21H40O2

IUPAC Name:

propyl oleate

Constituent 3

Reference substance name:

Propyl stearate

EC Number:

222-855-0

EC Name:

Propyl stearate

Cas Number:

3634-92-2

Molecular formula:

C21H42O2

IUPAC Name:

propyl stearate

Constituent 4

Reference substance name:

Heptadecanoic acid, propyl ester

Cas Number:

propyl heptadecanoate

Molecular formula:

C20H40O2

IUPAC Name:

Heptadecanoic acid, propyl ester

Constituent 5

Reference substance name:

Propyl (9Z)-9-hexadecenoate

Molecular formula:

C19H36O2

IUPAC Name:

Propyl (9Z)-9-hexadecenoate

Constituent 6

Reference substance name:

Propyl palmitate

EC Number:

218-803-1

EC Name:

Propyl palmitate

Cas Number:

2239-78-3

Molecular formula:

C19H38O2

IUPAC Name:

propyl palmitate

Constituent 7

Reference substance name:

Pentadecanoic acid, propyl ester

Molecular formula:

C18H36O2

IUPAC Name:

Pentadecanoic acid, propyl ester

Constituent 8

Reference substance name:

Propyl myristate

EC Number:

238-241-0

EC Name:

Propyl myristate

Cas Number:

14303-70-9

Molecular formula:

C17H34O2

IUPAC Name:

propyl myristate

Constituent 9

Reference substance name:

Propyl laurate

EC Number:

222-961-7

EC Name:

Propyl laurate

Cas Number:

3681-78-5

Molecular formula:

C15H30O2

IUPAC Name:

propyl laurate

Test material form:

liquid

Details on test material:

Name: 9-Octadecenoic acid (9Z)-, propyl ester
Product Description: Propyl oleate
CAS No.: 111-59-1
Physical state: colourless to yellow liquid at 20 °C
Batch No.: 37584
Re-certification date of batch: 08 March 2018
Purity: > 95 % (mono constituent substance, water content 0.33 % (w/w))
Free Fatty Acid, % Oleic: 0,1
Color, Gardner: 3
Iodine Value, cg I2/g 75
Cloud Point, degrees C 0
Saponification Number, mg KOH/g 177
Moisture, % 0,07
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

Specific details on test material used for the study:

Name: 9-Octadecenoic acid (9Z)-, propyl ester
Product Description: Propyl oleate
CAS No.: 111-59-1
Physical state: pale yellow to yellow liquid at 20 °C
Batch No.: 37584
Re-certification date of batch: 08 March 2018
Purity: > 99 % (fatty acid propyl esters, max water content 0.3 % (w/w))
Free Fatty Acid, % Oleic: 0.1
Color, Gardner: 3
Iodine Value, cg I2/g 75
Cloud Point, degrees C 0
Saponification Number, mg KOH/g 177
Moisture, % 0,07
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

Method

Target gene:

The Salmonella typhimurium histidine (his) reversion system measures his- → his+ reversions. The S. typhimurium strains are constructed to differentiate between base pair (TA 100, TA 1535, TA 102) and frameshift (TA 98, TA 1537) mutations.

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 Mix

Vehicle / solvent:

The test item was dissolved in ethanol and diluted prior to treatment. The solvent was compatible with the survival of the bacteria and the S9 activity.

Controls

Untreated negative controls:

yes

Remarks:

A. dest.

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 1. 4-NOPD; 4-nitro-o-phenylene-diamine; 2. 2-AA; 2-aminoanthracene

Evaluation criteria:

Evaluation of Mutagenicity

The Mutation Factor is calculated by dividing the mean value of the revertant counts by the mean values of the solvent control (the exact and not the rounded values are used for calculation).

A test item is considered as mutagenic if:

- a clear and dose-related increase in the number of revertants occurs and/or
- a biologically relevant positive response for at least one of the dose groups occurs

in at least one tester strain with or without metabolic activation.

A biologically relevant increase is described as follows:

- if in tester strains TA 98, TA 100 and TA 102 the number of reversions is at least twice as high
- if in tester strains TA 1535 and TA 1537 the number of reversions is at least three times higher

than the reversion rate of the solvent control.

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary. A test item producing neither a dose related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups is considered to be non-mutagenic in this system.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

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

Remarks:

Ethanol

Untreated negative controls validity:

valid

Remarks:

A. dest.

Positive controls validity:

valid

Remarks:

NaN3 (-S9); 2-AA (+S9)

Test results 2

Key result

Species / strain:

S. typhimurium 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

Remarks:

ethanol

Untreated negative controls validity:

valid

Remarks:

A. dest.

Positive controls validity:

valid

Remarks:

NaN3 (-S9); 2-AA (+S9)

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

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

Remarks:

ethanol

Untreated negative controls validity:

valid

Remarks:

A. dest.

Positive controls validity:

valid

Remarks:

NaN3 (-S9); 2-AA (+S9)

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

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

Remarks:

ethanol

Untreated negative controls validity:

valid

Remarks:

A. dest.

Positive controls validity:

valid

Remarks:

4-NOPD (-S9); 2-AA (+S9)

Test results 5

Key result

Species / strain:

S. typhimurium TA 102

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

Remarks:

ethanol

Untreated negative controls validity:

valid

Remarks:

A. dest.

Positive controls validity:

valid

Remarks:

MMS (-S9); 2-AA (+S9)

Remarks on result:

other: Experiment I (Plate-incorporation Test) & Experiment II (Plate-incorporation Test)

Any other information on results incl. tables

Discussion

The test item Propyl oleate was investigated for its potential to induce gene mutations according to the plate incorporation test (experiment I and II) using Salmonella typhimurium strains TA 98,TA 100, TA 1535, TA 1537 and TA 102.

In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item were prepared and used in the experiments:

Experiment I: 0.0316, 0.100, 0.316, 1.0, 2.5 and 5.0 µL/plate

Experiment II: 0.0158, 0.050, 0.158, 0.50, 1.58 and 5.0µL/plate

No precipitation of the test item was observed in any tester strain used in experiment I and II (with and without metabolic activation).

No toxic effects of the test item were noted in any of the five tester strains used up to the highest dose group evaluated with and without metabolic activation in experiment I and II. No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with Propyl oleate at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II. All criteria of validity were met.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, Propyl oleate did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, Propyl oleate is considered to be non-mutagenic in this bacterial reverse mutation assay.

Applicant's summary and conclusion

Conclusions:

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, Propyl oleate did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, Propyl oleate is considered to be non-mutagenic in this bacterial reverse mutation assay.

Executive summary:

The test item Propyl oleate was investigated for its potential to induce gene mutations according to the plate incorporation test (experiment I and II) using Salmonella typhimurium strains TA 98,TA 100, TA 1535, TA 1537 and TA 102.

In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with Propyl oleate at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II. All criteria of validity were met. In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, Propyl oleate did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, Propyl oleate is considered to be non-mutagenic in this bacterial reverse mutation assay.