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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
The substance has been tested in a guideline bacterial reverse mutation assay and found to be negative (not mutagenic). Other members of the C8-12 Alkenyl Succinic Anhydride category have also been nonmutagenic in in vitro genotoxicity tests. The conclusion is that the substance is not genotoxic.
Link to relevant study records
Reference
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:
other: Guideline study under GLP.
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
Target gene:
Histidine
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, and E. coli WP2.
1.0 to 5000 µg/plate for TA 1535 and TA 1537.
Vehicle / solvent:
DMSO
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
For all strains with S9
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylene-diamine
Remarks:
TA 98 and TA 1537 without S9
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
E. coli WP2 without S9
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 or 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.
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
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative

Tetrapropenyl Sussinic 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.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Tetrapropenyl succinic anhydride (TPSA) has been tested in a guideline bacterial reverse mutation assay (both plate incorporation method and preincubation method) and found to be negative (not mutagenic). Other members of the C8-12 Alkenyl Succinic Anhydride category (n-dodecenyl succinic anhydride, octenyl succinic anhydride, and tripropenyl succinic anhydride) have also been nonmutagenic in the Ames assay.  Tripropenyl succinic anhydride was tested in a guideline chromosomal aberrations assay and mammalian mutation assay (mouse lymphoma assay), and was found to be negative. An in vivo Unscheduled DNA Synthesis (UDS) assay on tripropenyl succinic anhydride was negative. In the review by the World Health Organization of Cyclic Acid Anhydrides (CICAD 75, 2009), genotoxicity tests for a variety of category members were negative for genotoxicity.  These data indicate that the alkenyl succinic anhydrides are not genotoxic.

A category approach is used for the hazard assessment of several endpoints. The hypothesis for the category of C8-12 Alkenyl Succinic Anhydrides is that data can be read-across among members of the category because their properties and behaviours are similar, based on common functional groups and similar breakdown products, and based on a constant pattern in changing of the potency of properties of the various carbon chain lengths. Functional groups include a dihydro-2,5 -furandione cyclic anhydride ring, a carbon chain of length 8 -12 carbons, and a single double-bond within the carbon chain. The primary functional group associated with toxicity is the succinic anhydride moiety, which quickly is hydrolysed to form a butanedioic acid. A constant pattern may also be displayed in acute toxicity, dermal irritancy and biodegradation, with the lowest carbon chain length (C8) displaying the highest activity. Irritation, toxicity and degradation potential diminish with increasing carbon chain length. Read-across among the category members is substantiated by the common behaviour in physico-chemical and toxicity behaviours, as provided in the Chemical Category Report Format (CCRF) attached to the IUCLID file. It is adequate to fulfill the information requirements of Annex IX, to be the basis for classification and labelling decisions, and for risk assessment. 


Justification for selection of genetic toxicity endpoint
Guideline study under GLP

Justification for classification or non-classification

This substance does not meet the criteria for mutagenicity in Regulation EC No. 1272/2008.