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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

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
Study period:
2010-09-29 and 2010-11-19
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-conform and according to guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1997
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of trans-5-hexyldihydro-4-methylfuran-2(3H)-one and cis-5-hexyldihydro-4-methylfuran-2(3H)-one
EC Number:
922-963-4
Molecular formula:
C11H20O2
IUPAC Name:
Reaction mass of trans-5-hexyldihydro-4-methylfuran-2(3H)-one and cis-5-hexyldihydro-4-methylfuran-2(3H)-one

Method

Target gene:
Salmonella typhimurium: histidine operon
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:
Phenobarbital/b-naphthoflavone induced rat liver S9
Test concentrations with justification for top dose:
- Pre-Experiment and Experiment I (plate incorporation test) : 3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate
- Experiment II (pre-incubation test): 1; 3; 10; 33; 100; 333; 1000; and 2500 µg/plate
Vehicle / solvent:
- Vehicle/solvent used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative non-toxicity to the bacteria.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: - S9: sodium azide (TA1535, TA100), 4-nitro-o-phenylene-diamine (TA1537, TA98), methyl methane sulfonate (WP2 uvrA); + S9: 2-aminoanthracene (TA 1535, TA 1537, TA 98, TA 100, WP2 uvrA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 60 minutes (pre-incuabtion method)
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of spontaneous revertants or a clearing of the bacterial background lawn


Evaluation criteria:
- A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
- A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
- An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
- A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.

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
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: The test item precipitated in the overlay agar in the test tubes from 2500 µg/plate up to 5000 µg/plate in the presence of metabolic activation. Precipitation on the incubated agar plates was observed from 1000 µg/plate up to 5000 µg/plate in the presence of metabolic activation in experiment I.

RANGE-FINDING/SCREENING STUDIES: In the pre-experiment the concentration range of the test item was 3 – 5000 µg/plate. The following concentrations were used in the main study: 1; 3; 10; 33; 100; 333; 1000; and 2500 µg/plate

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

ADDITIONAL INFORMATION ON CYTOTOXICITY: see "any other information on results"
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

The plates incubated with the test item showed reduced background growth at the following concentrations (µg/plate):

Strain

Experiment I

Experiment II

 

Without S9

With S9

Without S9

With S9

TA 1535

1000 - 5000

/

333 - 2500

1000 - 2500

TA 1537

333 - 5000

1000 - 5000

333 - 2500

1000 - 2500

TA 98

333 - 5000

/

100 - 2500

1000 - 2500

TA 100

333 - 5000

/

333 - 2500

1000 - 2500

WP2 uvrA

333 - 5000

/

1000 - 2500

1000 - 2500

Toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in the test groups at the following concentrations (µg/plate):

Strain

Experiment I

Experiment II

 

Without S9

With S9

Without S9

With S9

TA 1535

1000 - 5000

1000 - 5000

333 - 2500

1000 - 2500

TA 1537

333 - 5000

1000 - 5000

333 - 2500

1000 - 2500

TA 98

/*

2500 - 5000

100 - 2500

1000 - 2500

TA 100

2500 - 5000

2500 - 5000

1000 - 2500

1000 - 2500

WP2 uvrA

1000 - 5000

1000 - 5000

1000 - 2500

1000 - 2500

/* = no analysis possible at 5000 µg/plate

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under the experimental conditions, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Executive summary:

This study was performed to investigate the potential of the test substance to induce gene mutations in the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100, and the Escherichia coli strain WP2 uvrA with an without liver microsomal activation. The test item was tested at the following concentrations: 3, 10, 33, 100, 333, 1000, 2500, and 5000 µg/plate (pre-experiment/experiment I); 1, 3, 10, 33, 100, 333, 1000, and 2500 µg/plate (experiment II). Reduced background growth in the test groups with and without metabolic activation in both independent experiments was observed at higher concentrations. Toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in all strains. No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Peacholide at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies. In conclusion, these results indicated that the test substance under the experimental conditions described, was not mutagenic to Salmonella typhimurium strains and Escherichia coli in the absence and presence of a metabolizing system.