2-Octen-1-ylsuccinic anhydride, mixture of cis and trans

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-(oct-2-en-1-yl)dihydrofuran-2,5-dione

EC Number:

629-679-7

Cas Number:

42482-06-4

Molecular formula:

C12H18O3

IUPAC Name:

3-(oct-2-en-1-yl)dihydrofuran-2,5-dione

Test material form:

other: Waxy solid

Details on test material:

Octenyl Succinic Anhydride (OSA), Gulf Bayport Chemicals, LP. Lot # 44514821. Purity 99.0%, expiry date: 21 December 2012

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

S9 liver microsomal fraction from male Sprague Dawley rats induced with PB and BNF, purchased from Trinova Biochem GmbH, Giessen, Germany.

Test concentrations with justification for top dose:

3.16 to 5000 µg/plate for TA 98, TA 100, TA 1535, and E. coli WP2.
5.0 to 5000 µg/plate for TA 1537.

Vehicle / solvent:

DMSO

Controls

Controls 1

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

For all strains with S9

Controls 2

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Remarks:

TA 98 and TA 1537 without S9

Controls 3

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

E. coli WP2 without S9

Controls 4

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100 and TA 1535 without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION:in agar (plate incorporation); and preincubation;
DURATION
- Preincubation period: 60 minutes
- Exposure duration: in agar is 48 hours


DETERMINATION OF CYTOTOXICITY
- Method: Clearing of diminution of the background lawn

Colonies were counted using a ProtoCOL counter (Meintrup DWS Laborgerate GmbH.) In addition, TA 1535 and TA 1537 were counted manually.

Evaluation criteria:

The Mutation Factor is calculated by dividing the mean value of the revertant counts through 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 occur at non-toxic doses of the test item, and/or
- a biologically relevant positive response for at least one of the dose groups occurs in at least one tester strain with our without metabolic activation.

A biologically relevant increase is described as follows:
- if in tester strains TA 98, TA 100 and E. coli WP2 uvrA the number of reversions is at least twice as high
- if in tester strains TA 1535 and TA 1537 the number of revisions is at least three times higher

as compared to the reversion rate of the solvent control (5).

According to the 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.

Statistics:

No data.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In both the plate incorporation and pre-incubation method there was no increase in the mutation rate in any strains with our without S9.

Remarks on result:

other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:

Octenyl Succinic Anhydride was tested in an Ames Assay (OECD 471) using both a plate incorporation method and pre-incubation method. No increase in mutation rate was seen. The test substance is not mutagenic under the conditions of this assay.