Registration Dossier

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
(Z)-[(octadec-9-enyloxy)methyl]oxirane
EC Number:
262-268-7
EC Name:
(Z)-[(octadec-9-enyloxy)methyl]oxirane
Cas Number:
60501-41-9
Molecular formula:
C21H40O2
IUPAC Name:
2-[(octadec-9-en-1-yloxy)methyl]oxirane
Details on test material:
- Name of test material (as cited in study report): Cremophor Vorstufe
- Degree of purity/composition: 51.9 area% (main component)
- Physical state: Yellowish liquid
- Lot/batch No.: PN2
- Storage condition of test material: Room temperature (N2 conditions)

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S-9 mix
Test concentrations with justification for top dose:
0, 20, 100, 500, 2500, and 5000 µg/plate (Standard Plate Test)
0, 4, 20, 100, 500, and 2500 µg/plate (Preincubation Test)
Vehicle / solvent:
- Vehicle/solvent used: Acetone
- Justification for choice of solvent/vehicle: Due to the limited solubility of the test substance in water, acetone was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical contral data are available.
Controls
Untreated negative controls:
yes
Remarks:
sterility control
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
other: 2-aminoanthracene, 4-nitro-o-phenylendiamine, N-methyl-N'-nitro-N-nitrosoguanidine
Remarks:
with metabolic activation: 2-aminoanthracene (all strains); without metabolic activation: N-methyl-N'-nitro-N-nitrosoguanidine (TA 1535, TA 100), 4-nitro-o-phenylendiamine (TA 98), 9-aminoacridine (TA 1537) and 4-nitroquinoline-N-oxide (E Coli WP2 uvrA)
Details on test system and experimental conditions:
METHOD OF APPLICATION:
- 1st Experiment (standard plate test with and without S-9 mix)
- 2nd Experiment: (preincubation test with and without S-9 mix)

DURATION
- Preincubation period: 20 min (preincubation test only)
- Exposure duration: 48 - 72 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY: decrease in the number of revertants, reduced his or trp background growth and reduction in the titer.
Evaluation criteria:
Generally, the experiment is to be considered valid if the following criteria are met:
- The number of revertant colonies in the negative controls was within the normal range of the historleal control data for each tester strain.
- The sterility controls revealed no indleation of bacterial contamination.
- The positive control articles both with and without S-9 mix induced a significant increase in the number of revertant colonies within the range of the historical control data.
- The titer of viable bacteria was >1E9/mL.

The test chemical is considered positive in this assay if a dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

A test substance is generally considered nonmutagenic in this test if the number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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
Species / strain:
E. coli WP2 uvr A
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:
- A very weak bacteriotoxic effect (slight decrease in the number of revertants, slight reduction in the titer) was occasionally observed depending an the strain and test conditions at >2500 µg/plate.
- Precpitation of the test substance was found from about 500 µg/plate onward.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative