(Z)-N-octadecyldocos-13-enamide

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:

guideline study with acceptable restrictions

Remarks:

GLP guideline study with acceptable restrictions. Although 5 strains were tested no strain for detection of cross-linking or oxidising mutagens was included.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

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

Constituent 1

Reference substance name:

(Z)-N-octadecyldocos-13-enamide

EC Number:

233-226-5

EC Name:

(Z)-N-octadecyldocos-13-enamide

Cas Number:

10094-45-8

Molecular formula:

C40H79NO

IUPAC Name:

(Z)-N-octadecyldocos-13-enamide

Constituent 2

Reference substance name:

stearyl erucamide

IUPAC Name:

stearyl erucamide

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 S-9

Test concentrations with justification for top dose:

50, 150, 500, 1500, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: hexane
- Justification for choice of solvent/vehicle: solubility of test compound in vehicle

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

hexane

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 without S-9 Migrated to IUCLID6: 80 µg/plate

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

hexane

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

TA 100 and TA 1535 without S-9 Migrated to IUCLID6: 3 µg/plate (TA 100), 5 µg/plate (TA 1535)

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

hexane

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 and TA 1538 without S-9 Migrated to IUCLID6: 1 µg/plate (TA 98) and 2 µg/plate (TA 1538)

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

hexane

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene: 0.5 µg/plate with TA 1538 and TA 98, 1 µg/plate with TA 100, and 2 µg/plate with TA 1535 and TA 1537.

Remarks:

all strains with S-9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 3 days

NUMBER OF REPLICATIONS: triplicates; independent repetition of complete test

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth; other: reduction of background growth

Evaluation criteria:

Positive: increase in revertant colony numbers of at least twice of those of the concurrent solvent controls, with some evidence of a positive dose-relationship, in two separate experiments, with any bacterial strain either in presence or absence of S9-mix. No statistical analysis is performed.
Negative: no reproducible increases in revertant colony numbers of at least 1.5 times of those of the concurrent solvent controls, at any dose level with any bacterial strain. No statistical analysis is performed.
Ambiguous: If the results fail to satisfy the criteria for a clear positive or negative response as described the following approach is taken: 1) Repeat tests with modified experimental method may be performed. These modifications include (but are not restricted to) the use of a narrower dose range than that already tested; the use of different levels of liver homogenate S-9 fraction in the S-9 mix. Should an increase in revertant colony numbers be observed which satisfies the 'positive' criteria the material is considered to be mutagenic. No statistical analysis is performed. 2) If no clear 'positive' response can be obtained the test data may be subjected to analysis to determine the statistical significance of any observed increases in revertant colony numbers. The statistical procedure used is normally analysis of variance followed by Student's t-test.

Statistics:

In case of ambiguous results analysis of variance followed by Student's t-test is performed. In case of unequivocally positive or negative results according to the described criteria, no statistical analysis is performed.

Results and discussion

Test results

Test results 1

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

Remarks:

Precipitation starting at 1500 µg/plate in test 1 and at 5000 µg/plate in test 2.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

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

Remarks:

Precipitation starting at 1500 µg/plate in test 1 and at 5000 µg/plate in test 2.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True 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

Remarks:

Precipitation starting at 1500 µg/plate in test 1 and at 5000 µg/plate in test 2.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True 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

Remarks:

Precipitation starting at 1500 µg/plate in test 1 and at 5000 µg/plate in test 2.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

Precipitation starting at 1500 µg/plate in test 1 and at 5000 µg/plate in test 2.

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: Starting at 1500 µg/plate with and without metabolic activation in all strains in test 1 and at 5000 µg/plate with and without activation in all strains in test 2.

RANGE-FINDING/SCREENING STUDIES:
Dose range finding tests were performed with 5, 50, 500 and 5000 µg/plate. No cytotoxicity was observed, therefore 5000 µg/plate was chosen as maximum concentration for main test.

ADDITIONAL INFORMATION ON CYTOTOXICITY: No cytotoxicity was observed.

Any other information on results incl. tables

Maximum mean number of revertants (Test with highest revertant numbers chosen):

	Mean revertant numbers ±SD
	without S-9		with S-9
Strain	Solvent control	Test substance (µg/plate)	Solvent control	Test substance (µg/plate)
TA 1535	10±2.6	11±3.1 (150)	13±1.5	13±3.0 (50)
TA 1537	12±2.5	11±2.6 (150)	9±1.4	11±2.0 (50)
TA 1538	11±1.5	11±1.5 (500)	10±1.7	15±1.5 (500)
TA 98	23±1.2	25±2.0 (500)	23±3.6	24±0.6 (500)
TA 100	75±5.1	87±6.1 (150)	93±5.5	90±9.1 (5000)

 

The concurrent positive controls demonstrated the validity of the test system.

No substantial increases in revertant colony numbers of any of the tester strains were observed following treatment with the test substance at any dose level, neither in the presence nor absence of metabolic activation.

Conclusion:

Under the conditions chosen the test substance was not mutagenic in bacteria.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative