Glycerides, C16-18 and C18-unsatd., deodorizer distillates

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not available

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to standard guidelines in compliance with GLP

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Glycerides, C16-18 and C18-unsatd.

EC Number:

266-948-4

EC Name:

Glycerides, C16-18 and C18-unsatd.

Cas Number:

67701-30-8

Molecular formula:

No generic molecular formula is available

IUPAC Name:

Glycerides, C16-18 and C18-unsatd.

Details on test material:

- Name of test material (as cited in study report): Pine nut oil. Under the SDA nomenclature, the name of this substance is 'Glycerides, C16-18 and C18-unsatd.'
- Substance type: Triglycerides of vegetable origin
- Analytical purity: Analysis for contamination and microbiological analysis were preformed
- Composition of test material, percentage of components: Pinolenic acid 15.3%, mono unsaturated fatty acid 26.5%, poly unsaturated fatty acids 66%, oleic acid 25.1%, linoleic acid 45.5%, saturated fatty acids 7.1%
- Lot/batch No.: 7284/20071217-076
- Storage condition of test material: 119 d at 15 °C or 4 d at room temperature

Method

Target gene:

No data

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

rat liver microsomal enzyme homogenate

Test concentrations with justification for top dose:

10, 33, 100, 333 and 1,000 µg/plate

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

for TA 1535

Positive control substance:

sodium azide

Remarks:

None Migrated to IUCLID6: 5 µg/plate dissolved in DMSO

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

for TA 98

Positive control substance:

2-nitrofluorene

Remarks:

None Migrated to IUCLID6: 20 µg/plate dissolved in DMSO

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

for TA 100

Positive control substance:

methylmethanesulfonate

Remarks:

None Migrated to IUCLID6: 650 µg/plate dissolved in DMSO

Controls 4

Untreated negative controls:

yes

Remarks:

ethanol

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

for WP2 uvrA

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

None Migrated to IUCLID6: 10 µg/plate dissolved in DMSO

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

For the bacterial test with metabolic activation: TA1535, TA98 and TA100

Positive control substance:

other: 1 µg/plate 2-aminoanthracene dissolved in DMSO

Remarks:

none

Controls 6

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

For the bacterial test with metabolic activation: TA1537 and TA100

Positive control substance:

other: 2.5 µg/plate 2-aminoanthracene dissolved in DMSO

Remarks:

None

Controls 7

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

For the bacterial test with metabolic activation: TA1537

Positive control substance:

other: 5 µg/plate 2-aminoanthracene dissolved in DMSO

Remarks:

None

Controls 8

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

For the bacterial test with metabolic activation: WP2uvrA

Positive control substance:

other: 10 µg/plate 2-aminoanthracene dissolved in DMSO

Remarks:

None

Details on test system and experimental conditions:

METHOD OF APPLICATION: In agar (plate incorporation)

Evaluation criteria:

The negative control data (number of spontaneous revertants per plate) should be within the laboratory historical range for each tester strain. The positive control chemicals, sodium azide for TA1535; 9-aminoacridine for TA1537, 2-nitrofluorene for TA98, methylmethanesulphonate for TA100 and 4-nitroquinoline N-oxide for WP2uvrA should produce positive responses in all tester strains, which are within the laboratory historical range documented for each positive control substance. Furthermore, the mean plate count should be at least three times the concurrent vehicle control group means.
A test substance was considered negative (not mutagenic) in the test if (a) The total number of revertants in tester strain TA100 was not greater than
two times the concurrent control, and the total number of revertants in the tester strains TA1535, TA1537, TA98 or WP2uvrA was not greater than three times the concurrent control, (b) The negative response was reproducible in at least one independently repeated
experiment.

Statistics:

Not applicable

Results and discussion

Test results

Test results 1

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

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: The negative and strain-specific positive control values were within the laboratory historical control data ranges.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

None

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the conditions of this test, the test substance was considered to be non-mutagenic.

Executive summary:

A study was conducted to determine the potential mutagenicity of ‘glycerides, C16-18 and C18-unsatd.’ (as pine nut oil). The study was performed according to the requirements of the OECD Guideline 471.

Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA- were treated with the test substance using the Ames plate incorporation method at five dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system (10% liver S9 in standard co-factors). The dose range was determined in a preliminary toxicity assay and was 3 to 5,000 μg/plate. The experiment was repeated using the same dose range as the range-finding test.

The test substance precipitated at the two highest concentrations tested. The test substance was, therefore, tested up to the third dose level of 1,000 μg/plate. No test substance precipitate was observed on the plates at any of the doses tested in either the presence or absence of S9-mix.

No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test substance, either with or without metabolic activation.

Under the conditions of this test, the test substance was considered to be non-mutagenic.