5-cyclohexadecen-1-one

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:

24 February - 27 June 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

See below deviations to procotol

Principles of method if other than guideline:

Inital protocol: The bacterial cultures were incubated in a shaking water bath for 8 hours at 37°C.
New: The bacterial cultures were incubated in a shaking water bath at 37°C for 3.5 hours in the pre-experiment and in experiment I (strains TA 98 and TA 100) and for 3 hours in the experiment I and experiment II.
Reason for the alteration: The bacterial density was reached at the incubation times stated above.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(E)-cyclohexadec-5-enone

Cas Number:

35951-24-7

Molecular formula:

C16H28O

IUPAC Name:

(E)-cyclohexadec-5-enone

Constituent 2

Reference substance name:

(Z)-cyclohexadec-5-enone

Cas Number:

21944-95-6

Molecular formula:

C16H28O

IUPAC Name:

(Z)-cyclohexadec-5-enone

Specific details on test material used for the study:

Name (as stated in the report): Velvione
Batch: 9000353015
Expiration date: January 26, 2002

Method

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

33, 100, 333, 1000, 2500 and 5000 microg/plate (No relevant toxic effects occurred up to the maximal concnetrations) in the pre experiment study.

Vehicle / solvent:

Water deionised and DMSO

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

Negative solvent / vehicle controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Concentration 10 microg/plate

Controls 2

Untreated negative controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

Negative solvent / vehicle controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylene diamine (4-NOPD)

Remarks:

Concentrations of 10 microg/plate (TA 98) and 50 microg/plate (TA 1537)

Controls 3

Untreated negative controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

Negative solvent / vehicle controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

5,0 microg/plate

Controls 4

Untreated negative controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

Negative solvent / vehicle controls:

yes

Remarks:

Concurrent untreated and solvent control were performed

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene, 2 AA

Remarks:

2.5 microg/plate and 10 microg/plate for TA 102 only

Results and discussion

Test results

Key result

Species / strain:

other: TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects were observed in strains Ta 1535, TA 1537, TA 98 (Experiment II only)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

In both experiment, toxic effects, evident as a reduction in the number of revertants, were observed in strains TA 1535, TA 1537 and TA 98 (experiment II only) at the higher concentrations with and wihtout metabolic activation. The plates incubated with the test item showed normal background growth up to 5000 microg/plate with and without S9 mix in all strains used.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Velvione at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with the increasing concentrations in the range below the generally acknowledge border of biological relevance.

Applicant's summary and conclusion

Conclusions:

In conclusion it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Therefore the VELVIONE is considered to be non mutagenic in this Samonella Typhimurium reverse mutation assay.VELVIONE does not met the criteria for mutagenic classication according the CLP Regulation (EC) No. 1272/2008.

Executive summary:

This study was performed to investigate the potential of Velvione to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella Typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102.

The assay was performed in two independant experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The tes item was tested at the following concentrations: 33; 100; 333; 1000; 2500; and 5000 microg/plate.

In both experiment, toxic effects, evident as a reduction in the number of revertants, were observed in strains TA 1535, TA 1537 and TA 98 (experiment II only) at the higher concentrations with and wihtout metabolic activation. The plates incubated with the test item showed normal background growth up to 5000 microg/plate with and without S9 mix in all strains used.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Velvione at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with the increasing concentrations in the range below the generally acknowledge border of biological relevance.

In conclusion it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

Therefore the VELVIONE is considered to be non mutagenic in this Samonella Typhimurium reverse mutation assay.VELVIONE does not met the criteria for mutagenic classication according the CLP Regulation (EC) No. 1272/2008.