Registration Dossier

Diss Factsheets

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:
26 Jun 2006 - 20 Jul 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
(2-ethyl-2-methyl-1,3-dioxolan-4-yl)methyl prop-2-enoate
EC Number:
807-159-2
Cas Number:
69701-99-1
Molecular formula:
C10H16O4
IUPAC Name:
(2-ethyl-2-methyl-1,3-dioxolan-4-yl)methyl prop-2-enoate
Test material form:
liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 19892501
- Purity: 97.5%

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Source: Dr. T. Matsushima, Japan Bioassay Laboratory, Japan Industrial Safety and Health Association, Hadano-city, Kanagawa.
Bacterial cultures were freshly prepared in Oxoid nutrient broth #2 (248458), by inoculating bacteria from frozen stock cultures (kept at -80 °C). 30µL stock solution was inoculated into the 15 mL broth and incubated for 10 hours at 37 °C and 120 rpm using rotary shaker.

S9 and S9 mix:
- Lot No: 06051202
- Source: Oriental Yeast Corporation, Tokyo, Japan.
- Date of preparation: 12 May 2006
- Preparation: prepared by a slight modification of the method described by Ames, McCann, and Yamazaki: 5,6-Benzoflavone (BF) and phenobarbital (PB) were used as an incuder of drug-metabolising enzyme system.

Animals:
- Species/ Strain: Rat/ Sprague-Dawley
- Sex: Male
- Age: 7 weeks
- Body weight: 217.5 ± 10.9 g
- Amount of inducing substance (g/kg bw): PB – 4 times, 0.03-0.06; BF – 1 time, 0.08

The S9 mix contained 4mM NADPH, 4mM NADH, 5mM Glucose-6-Phosphate, 8mM MgCl2, 33 mM KCl, 100mM sodium phosohate buffer (pH 7.4) and 10% (50µL S9 per plate).
Test concentrations with justification for top dose:
Range finder test (µg/ plate): 0, 4.88, 19.5. 78.1. 313, 1250 and 5000
Mutgenicity test (µg/ plate): 0, 156, 313, 625, 1250, 2500 and 5000
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility
Controls
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide); 2-aminoamthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION:
The mutagenicity was assayed from a maximum level of 5000 µg test substance/ plate by the pre-incubation method (37 °C, 20 min) with and without metabolic activation system.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 5000 µg/plate
Vehicle controls validity:
not specified
Positive controls validity:
not specified
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 2500 µg/plate
Vehicle controls validity:
not specified
Positive controls validity:
not specified
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 5000 µg/plate
Vehicle controls validity:
not specified
Positive controls validity:
not specified
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 5000 µg/plate
Vehicle controls validity:
not specified
Positive controls validity:
not specified
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 5000 µg/plate
Vehicle controls validity:
not specified
Positive controls validity:
not specified
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 5000 µg/plate
Vehicle controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
MEDOL-10 did not increase revertant colonies when tested up to 5000 µg/plate with or without metabolic activation.
Toxicity was observed at 5000µg/plate with or without metabolic activation for all bacteria tested.
Toxicity was also observed at 2500 µg/plate without metabolic activation for Salmonella Typhimurium TA100.
No test item precipitation was observed.
Remarks on result:
no mutagenic potential (based on QSAR/QSPR prediction)

Applicant's summary and conclusion

Conclusions:
Test item did not show mutagenicity in any bacteria strain by the preincubation method with or without metabilic activation system.