DODECANEDIOIC ACID, 1,12-BIS(2-OCTYLDODECYL) ESTER

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Based on the results of this ames study it is concluded that Dioctyldodecyl Dodecanedioate is not mutagenic in the Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and E. coli WP2 in the presence and absence of meta-bolic activation under the experimental conditions in this study.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05May2021 - 24Jun2021

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 26Jun2020

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

adopted 30 May 2008

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

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:

benzo(a)pyrene

Test concentrations with justification for top dose:

Plate incorporation method: 5, 1.5, 0.5, 0.15 and 0.05 μL/plate.
Pre-incubation method: 5, 2.5, 1.25, 0.63, 0.31 ,0.16 and 0.08 μg/plate.

Vehicle / solvent:

Acetone
Based on the non-GLP pre-test, acetone was chosen as vehicle, because the test item was sufficiently soluble, and this solvent does not have any effects on the viability of the bacteria or the number of spontaneous revertants in the tested concentrations.

Untreated negative controls:

yes

Remarks:

spontaneous revertant rates

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulfoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Without S9 mix for 20µg plate for TA98
Without S9 mix for 30µg plate for TA1537

Untreated negative controls:

yes

Remarks:

spontaneous revertant rates

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulfoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

With S9 mix for 20 µg/plate for TA98

Untreated negative controls:

yes

Remarks:

spontaneous revertant rates

Negative solvent / vehicle controls:

yes

Remarks:

Demin Water

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Without S9 mix for 1.8µg/plate for TA100
Without S9 mix for 1µg/plate for TA1535

Untreated negative controls:

yes

Remarks:

spontaneous revertant rates

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulfoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

With S9 mix for 1µg/plate for TA100
With S9 mix for 1µg/plate for TA1535
With S9 mix for 2.4µg/plate for TA1537
With S9 mix for 50µg/plate for E. Coli

Untreated negative controls:

yes

Remarks:

spontaneous revertant rates

Negative solvent / vehicle controls:

no

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

Without S9 mix for 2µg/plate for E. coli

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
Per bacteria strain and concentration, three plates with (+S9) and three plates without metabolic activation (-S9) were used.

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable):
- Test substance added in medium; in agar (plate incorporation);
preincubation;

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 20 mins
- Exposure duration/duration of treatment: 48 hrs
- Harvest time after the end of treatment (sampling/recovery times):


FOR GENE MUTATION:

- Number of cells seeded and method to enumerate numbers of viable and mutants cells: at least 109 bacteria/mL (correlating to 100 colo-nies/plate after dilution),
- Criteria for small (slow growing) and large (fast growing) colonies:

METHODS FOR MEASUREMENT OF CYTOTOXICITY
The bacterial background lawn was visible and not affected. The number of revertant colo-nies was not reduced. Thus, no signs of toxicity towards the bacteria strains could be ob-served.

METHODS FOR MEASUREMENTS OF GENOTOXICIY

- OTHER:
Sterility was examined

Evaluation criteria:

A biologically relevant increase is described as follows:
 if in the bacteria strains S. typhimurium TA98, TA100 and E. coli WP2 the number of revertants is at least twice as high than the reversion rate of the negative controls (increase factor of at least 2.0)
 if in the bacteria strains S. typhimurium TA1535 and TA1537 the number of re-vertants is at least three times higher than the reversion rate of the negative con-trols (increase factor of at least 3.0).

Statistics:

standard deviation

Key result

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:

valid

Positive controls validity:

valid

Key result

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:

valid

Positive controls validity:

valid

Key result

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:

valid

Positive controls validity:

valid

Key result

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:

valid

Positive controls validity:

valid

Key result

Species / strain:

E. coli WP2

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

True negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

Based on the results of this study it is concluded that Dioctyldodecyl Dodecanedioate is not mutagenic in the Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and E. coli WP2 in the presence and absence of meta-bolic activation under the experimental conditions in this study.

Executive summary:

Based on the results of this study it is concluded that Dioctyldodecyl Dodecanedioate is not mutagenic in the Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and E. coli WP2 in the presence and absence of meta-bolic activation under the experimental conditions in this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Justification for classification or non-classification

Based on the results of this ames study it is concluded that Dioctyldodecyl Dodecanedioate is not mutagenic in the Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and E. coli WP2 in the presence and absence of meta-bolic activation under the experimental conditions in this study.