Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
21 July 1997
Deviations:
no
Qualifier:
according to
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

Reference
Name:
Unnamed
Type:
Constituent
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.
Controlsopen allclose all
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
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
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
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
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
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
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
Key result
Species / strain:
other: Salmonella typhimurium TA97a, TA98, TA100, TA102 and TA1535
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
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.