3-(chlorodimethylsilyl)propyl methacrylate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28th Jan 2003 to 17th Feb 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

121177-93-3

Test material form:

liquid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature, protected from light
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: The test item was dissolved in DMSO. The solvent was compatible with the survival of the bacteria and S9 activity.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was dissolved in DMSO
- Preliminary purification step (if any): not performed
- Final dilution of a dissolved solid, stock liquid or gel: not applicable
- Final preparation of a solid: not applicabl

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA97a, TA98, TA100 and TA102

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital (80 mg/kg b.w.) and ß-Naphtoflavone (100 mg/kg b.w.) induced rat liver S9

Test concentrations with justification for top dose:

pre-experiment: 3.2, 10.2, 32.1, 101.5, 320.8, 1015.2, 2538, 5076 µg/plate. To determine the toxicity of the test item in TA98 and TA100
exposure concentrations: 32.1, 101.5, 320.8, 1015.2, 2538 and 5076 µg/plate. Concentrations based on results from the pre-experiment.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was compatible with the survival of the bacteria and S9 activity.

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylene-diamine, 4-NOPD

Remarks:

without metabolic activation, TA 97a, TA 98

Controls 2

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

without metabolic activation, TA 102

Controls 3

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

without metabolic activation, TA 1535, TA 100

Controls 4

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene, 2-AA

Remarks:

with metabolic activation, TA 1535, TA 97a, TA 98, TA 100 and TA 102

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation
- Cell density at seeding (if applicable): not specified

ACTIVATION: The protein concentration in the S9 preparation was 44.5 mg/ml. 100 mM sodium-ortho-phosphate buffer, pH 7.4 were ice-cold added to the following pre-weighed reagents to give final concentrations in the S9 mix of: 8 mM MgC12, 33 mM KCI, 5 mM Glucose-6-phosphate, 5 mM NADP.

DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells):

NUMBER OF REPLICATIONS: Triplicate

NUMBER OF CELLS EVALUATED: not specified

DETERMINATION OF CYTOTOXICITY
- Method: background lawn growth; number of revertants; mutation factor
- Any supplementary information relevant to cytotoxicity: not applicable

OTHER EXAMINATIONS:
- Determination of polyploidy: not applicable
- Determination of endoreplication: not applicable

Rationale for test conditions:

Concentrations based on results from the pre-experiment.

Evaluation criteria:

A test item was considered mutagenic when:
- a dose-related increase in the number of revertants occured and/or;
- a reproducible biologically relevant positive response for at least one of the dose groups occurs in at least one strain with or without metabolic activation

Statistics:

Not specified

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: TA1535, TA97a, TA100, TA98 and TA102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium, other: TA1535 and TA100

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 2538 µg/plate and 5076 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium, other: TA1535 and TA100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5076 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium, other: TA97a and TA102

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5076 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 2538 and 5076 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not specified
- Effects of osmolality: not specified
- Evaporation from medium: not specified
- Water solubility: not specified
- Precipitation: not specified
- Definition of acceptable cells for analysis: not specified
- Other confounding effects: not specified

RANGE-FINDING/SCREENING STUDIES: No mutagenic or cytotoxic effects were observed.

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: Results within historical control data ranges.
- Negative (solvent/vehicle) historical control data: Results within historical control data ranges.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: background lawn
- Other observations when applicable: not applicable

Remarks on result:

other:

Remarks:

Pre-experiment

Any other information on results incl. tables

Table 1: Experiment I, plate incorporation test

Concentration μg/plate	TA1535		TA98		TA97a		TA100		TA102
	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA
A dest.	24	25	23	20	105	130	117	103	274	263
DMSO	23	23	24	21	106	97	101	101	269	277
32.1	25	27	22	18	111	112	117	88	405	333
101.5	23	26	23	14	124	108	101	82	319	355
320.8	22	26	21	21	105	105	117	95	413	219
1015.2	22	23	19	15	140	123	114	85	431	396
2538	26	26	16	15	115	101	93	89	436	434
5076	25	25	17	20	103	107	101	90	286	280
NaN3 10	/	519	/	/	1364	/	/	1166	/	/
2-AA 2.5	261	/	2055	/	/	/	2297	/	669	/
4-NOPD 40	/	/	/	/	/	1036	/	/	/	/
4-NOPD 10	/	/	/	863	/		/	/	/	/
MMS 1200	/	/	/	/	/	/	/	/	/	1035

Table 2: Experiment II, pre-incubation test

Concentration μg/plate	TA1535		TA98		TA97a		TA100		TA102
	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA
A dest.	10	12	31	26	140	91	120	106	384	364
DMSO	11	10	26	23	166	118	104	96	346	361
32.1	9	18	33	20	89	93	109	123	309	227
101.5	11	20	27	20	117	112	119	113	323	272
320.8	11	15	29	23	135	113	107	122	348	295
1015.2	12	14	27	26	163	140	112	100	327	283
2538	9	14	21	32	167	137	101	88	268	269
5076	11	12	33	22	133	99	119	177	357	209
NaN3 10	/	1185	/	/	/	/	/	730	/	/
2-AA 2.5	193	/	2783	/	1459	/	1632	/	742	/
4-NOPD 40	/	/	/	/	/	1261	/	/	/	/
4-NOPD 10	/	/	/	1165	/	/	/	/	/	/
MMS 1200	/	/	/	/	/	/	/	/	/	2422

Applicant's summary and conclusion

Conclusions:

[Dimethoxy(methyl)silyl]methyl methacrylate (CAS No 121177-93-3) has been tested in a valid bacterial reverse mutation assay, according to the OECD TG 471 (1989), and under GLP, using Salmonella typhimurium strains TA 1535, TA 97a, TA 98, TA 100 and TA 102. No increase in the number of revertants was observed in any test strain, with or without metabolic activation. Appropriate positive, negative and solvent controls were added and gave expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.