(Z)-N-octadec-9-enylhexadecan-1-amide

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:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study with acceptable restrictions. Although 5 strains were tested no strain for detection of cross-linking or oxidising mutagens was included, but in accordance with guideline at time of testing.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Strain for detection of oxidising or cross-linking mutagens is missing, but in accordance with guideline at the time of testing.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

(Z)-N-octadec-9-enylhexadecan-1-amide

EC Number:

240-367-6

EC Name:

(Z)-N-octadec-9-enylhexadecan-1-amide

Cas Number:

16260-09-6

Molecular formula:

C34H67NO

IUPAC Name:

N-octadec-9-en-1-ylhexadecanamide

Constituent 1

Reference substance name:

(Z)-N-octadec-9-en-1-ylhexadecanamide

Molecular formula:

C34H67NO

IUPAC Name:

(Z)-N-octadec-9-en-1-ylhexadecanamide

Constituent 2

Reference substance name:

oleyl palmitamide

IUPAC Name:

oleyl palmitamide

Test material form:

other: solid, not further specified

Method

Target gene:

His operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98, TA 100

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9 mix

Test concentrations with justification for top dose:

100, 333, 1000, 3330, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Very low water solubility of test substance in common solvents. Appropriate solvents not suitable for bacterial culture; therefore, suspension of test substance in DMSO added to cultures.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 1535 without S9 mix, 1 µg/plate

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA 100 without S9 mix, 0.5 µL/plate

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylene-diamine (4 NPD), 10 µg/plate

Remarks:

TA 98 and TA 1538 without S9 mix

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 without S9 mix; 60 µg/plate

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2AA), 0.5 µg/plate

Remarks:

All strains with S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: triplicates

DETERMINATION OF CYTOTOXICITY
- Method: reduction of background growth

Evaluation criteria:

Acceptability of Assay:
1) Negative control data (number of spontaneous revertants per plate) should reasonably fall within the laboratory background historical range for each tester strain.
2) The positive control chemicals should produce responses in all tester strains which also reasonably fall within the laboratory historical range documented for each positive control substance.
3) The selected dose range should include a clearly toxic concentration as demonstrated by a preliminary toxicity range-finding test with strain TA 100.

Statistics:

Mean and standard deviation of revertant numbers.

Results and discussion

Test results

Test results 1

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

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

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

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1538

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

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

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

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

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

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation occurred up to limit concentrations.

RANGE-FINDING/SCREENING STUDIES:
Eleven serial dilutions of the test substance (0.1, 0.3, 1.0, 3.3, 10.0, 33.3, 100, 333, 1000, 3330, 5000 µg/plate) were plated with a diluted TA 100 culture on non-selective agar for viability counting. For viability counting equal numbers of bacterial cells were plated in the presence of test substance. The percentage of survival of an appropriately diluted TA 100 culture on non-selective agar was determined by comparing the number of colonies on the solvent control plate with those on the test substance plates. Even at the highest test substance concentration used (5000 µg/plate) there was no reduction in survival of TA 100. Therefore, the test substance was tested up to a concentration of 5000 µg/plate in the main test, which is the maximum test concentration to be used according to the OECD guideline.

COMPARISON WITH HISTORICAL CONTROL DATA:
The observed numbers of revertants are well within the ranges of the historical controls.

Any other information on results incl. tables

Maximum mean numbers of revertants (± SD):

Strain	-S9		+S9
	Control	Test substance (µg/plate)	Control	Test substance (µg/plate)
TA 1535	9 ± 2	9 ± 1 (100)	12 ± 4	14 ± 2 (333)
TA 1537	12 ± 4	14 ± 3 (1000)	9 ± 2	20 ± 19 (5000)
TA 1538	14 ± 2	19 ± 6 (100)	26 ± 6	28 ± 3 (100)
TA 98	36 ± 10	38 ± 9 (333)	30 ± 14	36 ± 8 (333)
TA 100	121 ± 6	119 ± 7 (5000)	98 ± 19	138 ± 9 (5000)

 

The test substance did not induce a statistically significant dose-related increase in the numbers of revertants in any of the tester strains.

Conclusion:

The test substance does not have to be considered as mutagenic to bacteria.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative