(2-ethyl-2-methyl-1,3-dioxolan-4-yl)methyl prop-2-enoate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Bacterial reverse mutation, in vitro - Negative with and without metabolic activation.

Human Lymphocytes micronucleus assay, in vitro - Mutagenic with and without metabolic activation

Chromosome aberration, in vitro -

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

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%

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

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.

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)

Conclusions:

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

Endpoint conclusion

Endpoint conclusion:

no study available (further information necessary)

Additional information

A Bacterial reverse mutation test was conducted (UBE Scientific Analysis Laboratory, Inc, 2006, Study USA-R-06344) to examine the potential for MEDOL-10 to cause gene mutation. The study was conducted according Japan Guidelines for Screening Mutagenicity Testing Of Chemicals.

 

The strains of Salmonella Typhimurium (TA1535, TA1537, TA98, and TA100) and Escherichia Coli WP2uvrA were exposed to MEDOL-10 as a solution in Dimethylsulphoxide (DMSO) using the pre-incubation methods at up to six dose levels, in duplicate in both the presence and absence of metabolic activation. The six concentrations of MEDOL-10 were 156, 313, 625, 1250, 2500 and 5000µg/plate.

 

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 during the experiment.

 

In the Reverse Mutation Assay using strains of Salmonella typhimurium and Escherichia coli MEDOL-10 did not induce a significant increase in the frequency of revertant colonies at any of the dose levels used either with or without metabolic activation. Under the conditions of MEDOL-10 was considered to be non-mutagenic.

The study was performed (Covance Laboratories Limited, 2020, Study LB03SD) to detect the clastogenic and aneugenic potential of MEDOL-10 on the nuclei of normal human lymphocytes. The study was performed in accordance to OECD test guideline 487 and in compliance with GLP.

Duplicate cultures of human lymphocytes were evaluated for micronuclei in binucleate cells at up to four dose levels, together with quadruplicate vehicle and duplicate positive controls cultures. Three exposure conditions in a single experiment were used: a 4-hour exposure in the presence and absence of a standard metabolizing system (S9) at a 2% final concentration, and a 24-hour exposure in the absence of metabolic activation. At the end of the exposure period, the cell cultures were washed and then incubated for a further 24 hours in the presence of Cytochalasin B.

There were binucleate cells present up to 125 μg/mL in the 4 hour exposure in the absence of S9, up to 800 μg/mL in the presence of S9 and up to 54.69 μg/mL in the 24-hour exposure group. The CBPI data for the 4-hour exposure groups in the absence and presence of S9 and for the 24-hour confirm the qualitative observations in that a dose-related toxicity was observed in all three exposure groups.

MEDOL-10 induced statistically significant dose related increases in the frequency of binucleate cells with micronuclei in both the absence and presence of a metabolizing system. The test item was considered to be mutagenic to human lymphocytes in vitro.