Dihydro-3-[3-(triethoxysilyl)propyl]furan-2,5-dione

Genetic toxicity: in vitro

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:

2002-08-20 to 2002-09-06

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

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:

3-[3-(triethoxysilyl)propyl]oxolane-2,5-dione

IUPAC Name:

3-[3-(triethoxysilyl)propyl]oxolane-2,5-dione

Constituent 2

Reference substance name:

Dihydro-3-[3-(triethoxysilyl)propyl]furan-2,5-dione

EC Number:

297-566-6

EC Name:

Dihydro-3-[3-(triethoxysilyl)propyl]furan-2,5-dione

Cas Number:

93642-68-3

Molecular formula:

C13H24O6Si

IUPAC Name:

3-[3-(triethoxysilyl)propyl]dihydrofuran-2,5-dione

Test material form:

other: liquid

Method

Target gene:

histidine operon (Salmonella strains); tryptophan operon (E.coli strain)

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

Experiment 1: 3, 10, 33, 100, 333, 1000, 3330, 5000 μg/plate. Experiment 2: 100, 333, 1000, 2000, 3330, 5000 μg/plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

- Justification for choice of solvent/vehicle: Solubility properties and relative non-toxicity to bacteria

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 1535 (without activation) 1 μg/plate

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 (without activation) 60 μg/plate

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: daunomycine

Remarks:

TA 98 (without activation) 4 μg/plate

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA 100 (without activation) 650 μg/plate

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

WP2 uvrA (without activation) 10 μg/plate

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

All strains (with activation) TA 1537 2.5 μg/plate; TA 1535, TA 98 & TA100 1 μg/plate; WP2 uvrA 5 μg/plate

Details on test system and experimental conditions:

ACTIVATION: S9-mix contained per 10 mL:
- 30 mg NADP
- 15.2 mg g-6-p in 5.5 or 5.0 mL Milli-Q water
- 2 mL 0.5 M sodium phosphate buffer pH 7.4
- 1 mL 0.08 M MgCl2
- 1 mL 0.33 M KCl
In experiment 1, 0.5 mL S9-fraction was added (5% (v/v)) was added to 9.5 mL of S9-mix components. 0.5 ml S9 mix was added to 3.2 ml top agar with test substance and bacterial giving a final concentration of approximately 0.7% S9
In experiment 2, 1.0 mL S9-fraction was added (10% (v/v)) was added to 9.0 mL of S9-mix components. 0.5 ml S9 mix was added to 3.2 ml top agar with test substance and bacterial giving a final concentration of approximately 1.4% S9

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Expression time (cells in growth medium): 48 hours at 37°C

NUMBER OF REPLICATIONS: 3 plates for each test concentration; the experiment was repeated, and a third experiment carried out with TA100 without metabolic activation.

DETERMINATION OF CYTOTOXICITY
- Method: Background lawn assessment, revertant colony counts.

Evaluation criteria:

For a test substance to be considered positive, it must cause at least a doubling in the mean revertants per plate of at least one tester strain.

The increase must be accompanied by a dose response towards increasing concentrations of the test article.

The positive response should be reproducible in at least one independently repeated experiment.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

3330 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 6

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

3330 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 7

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 8

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

3330 μg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 9

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: Results were within range of historical control data

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Experiment 1: Number of revertants (Mean of 3 plates)

Concentration (µg/plate)	TA 98		TA 100		TA 1535		TA 1537		WP2 uvrA
	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA
DMSO	14	20	116	127	10	13	9	10	20	22
3			113	138					23	24
10			99	130					25	20
33	17		137	109	10		5	11	24	27
100	12	17	127	110	11	12	7	7	25	26
333	15	18	139	107	11	7	11	10	21	18
1000	13	19	133	102	10	8	5	10	27	22
3330	7**	17	37**	101	MC****	7	MC****	8	15	26
5000		14	0****	86*		5	347	435	13*	19
Positive control	588	868	679	804	644	218			455	100

MC = microcolonies

*Bacterial background lawn slightly reduced

**Bacterial background lawn moderately reduced

***Bacterial background lawn extremely reduced

****Bacterial background lawn absent

Experiment 2: Number of revertants (Mean of 3 plates)

Concentration (µg/plate)	TA 98		TA 100		TA 1535		TA 1537		WP2 uvrA
	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA
DMSO	17	18	119	81	10	6	6	6	17	19
100	14	20	74	84	11	9	7	6	16	19
333	15	19	135	149	14	7	8	8	14	17
1000	16	19	122	68	5	11	5	6	15	19
2000	11	-	104	-	8*	-	5	-	-	-
3330	12*	17	73	80	4*	9	2*	6	14	19
5000	-	18	-	83	-	7	-	6	13	15
Positive control	263	278	715	406	325	64	346	60	288	137

*Bacterial background lawn slightly reduced

Experiment 3: Number of revertants (Mean of 3 plates)

Concentration (µg/plate)	TA 100
	-MA
DMSO	101
100	127
333	118
1000	106
3330	71*
5000	MC***
Positive control	433

MC = microcolonies

*Bacterial background lawn slightly reduced

***Bacterial background lawn extremely reduced

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

3-[3-(Triethoxysilyl)propyl]oxolane-2,5-dione has been tested for mutagenicity to bacteria, in a study which was conducted according to the OECD TG 471, in compliance with GLP. No evidence of a test-substance related increase in the number of revertants was observed with or without metabolic activation in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 or E. coli WP2 uvrA in the initial or the repeat experiments up to cytotoxic or limit concentrations. Appropriate positive and solvent controls were included and gave the expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.