Ditetradecyl peroxydicarbonate

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:

2011-09-22 to 2011-10-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

30 May 2008

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ditetradecyl peroxydicarbonate

EC Number:

258-436-4

EC Name:

Ditetradecyl peroxydicarbonate

Cas Number:

53220-22-7

Molecular formula:

C30H58O6

IUPAC Name:

1-[({[(tetradecyloxy)carbonyl]peroxy}carbonyl)oxy]tetradecane

Test material form:

solid

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97a, TA98, TA100, TA102 and TA1535

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9 mix

Test concentrations with justification for top dose:

In the first experiment:
21, 62, 185, 556 and 1667 μg per plate without external metabolisation, and
21, 62, 185, 556 and 1667 μg per plate with S9-mix from Aroclor 1254 induced microsomes of rat liver as an external metabolising system.
In the second experiment:
7, 21, 62, 185 and 556 μg per plate without external metabolisation, and
7, 21, 62, 185 and 556 μg per plate with S9-mix from Aroclor 1254 induced microsomes of rat liver as an external metabolising system.

Vehicle / solvent:

Acetone
- Vehicle/solvent used: acetone
- Justification for choice of solvent/vehicle: The test substance was not soluble in water or DMSO. Acetone is a common vehicle for the Ames test.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

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

Remarks:

TA 97a, without S9 mix

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100, TA 1535 without S9 mix

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminonthracene (2-AA)

Remarks:

TA 98, TA 100, TA 1535 with S9 mix

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 without S9 mix

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

Remarks:

TA 97a with S9 mix

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 1,8-dihydroxyanthraquinone (DHA)

Remarks:

TA 102 with S9 mix

Controls 7

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: t-butyl-hydroperoxide (tBHPO)

Remarks:

TA 102 without S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation;

DURATION
- Preincubation period: 20 min
- Exposure duration: 2 days

NUMBER OF REPLICATIONS: 3 per dose and positive controls; 6 per vehicle control

DETERMINATION OF CYTOTOXICITY
- Method: reduced or no bacterial background lawn, cloning efficiency

Evaluation criteria:

Means and standard deviations were calculated for the number of mutants in every concentration group.
The criteria for a positive result are:
A reproducible increase of the number of revertants to more than the following threshold values for at least one of the concentrations:
• For the strains with a low spontaneous revertant rate i.e. TA98 and TA1535: The 2½ fold of the amount of the spontaneous revertants.
• For the strains with a high spontaneous revertant rate i.e. TA97a, TA100 and TA102: The 1 2/3 fold of the amount of the spontaneous revertants.
These threshold values were derived from the variations in the control samples of our historic data of the Ames test.

Statistics:

NA

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 97a

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

(1667 µg/plate)

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 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

(1667 µg/plate)

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:

(1667 µg/plate)

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 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

(1667 µg/plate)

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 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

(1667 µg/plate)

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
- Water solubility: no
- Precipitation: at 1667 µg/plate
- Other confounding effects: none

RANGE-FINDING/SCREENING STUDIES: In the preliminary test and in the main test no toxicity was seen up to 1667 μg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: yes

Applicant's summary and conclusion

Conclusions:

Dimyristylperoxydicarbonate is nonmutagenic in the Ames test with the strains TA 97a, TA 98, TA 100, TA 102 and TA 1535 with and without an external metabolising system up to 1667 μg/plate, which is the limit of solubility.

Executive summary:

The test item Dimyristylperoxydicarbonate was tested for mutagenic activity with the "Salmonella typhimurium Reverse Mutation Test" (Ames Test).

The study was conducted in accordance with the OECD-guideline 471 and the Council Regulation (EC) No 440/2008, Method B.13/14. The test substance was dissolved in acetone.

The following concentrations were tested:

In the first experiment:

21, 62, 185, 556 and 1667 μg per plate without external metabolisation, and

21, 62, 185, 556 and 1667 μg per plate with S9-mix from Aroclor 1254 induced microsomes of rat liver as an external metabolising system.

In the second experiment:

7, 21, 62, 185 and 556 μg per plate without external metabolisation, and

7, 21, 62, 185 and 556 μg per plate with S9-mix from Aroclor 1254 induced microsomes of rat liver as an external metabolising system.

In the first experiment the test was performed according to the "direct plate incorporation method", in the second experiment according to the "preincubation method".

As test system the bacterial strains Salmonella typhimurium TA 97a, TA 98, TA 100, TA 102 and TA 1535 were used.

Negative and positive controls were included.

According to the results obtained in this study, Dimyristylperoxydicarbonate is nonmutagenic in the Ames test with the strains TA 97a, TA 98, TA 100, TA 102 and TA 1535 with and without an external metabolising system up to 1667 μg/plate, which is the limit of solubility.