N-(3-(trimethoxysilyl)propyl)ethylenediamine

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:

1998-11-10 - 1998-11-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-(3-(trimethoxysilyl)propyl)ethylenediamine

EC Number:

217-164-6

EC Name:

N-(3-(trimethoxysilyl)propyl)ethylenediamine

Cas Number:

1760-24-3

Molecular formula:

C8H22N2O3Si

IUPAC Name:

N-(3-(trimethoxysilyl)propyl)ethylenediamine

Method

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, other: E coli WP2 trp pKM101 (CM881)

Species / strain 3

Species / strain / cell type:

E. coli, other: E coli WP2 trp uvrA pKM101 (CM891)

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9

Test concentrations with justification for top dose:

50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: Sponsors choice as completely miscible in water

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

without metabolic activation TA1535 5 µg/plate, TA100 3 µg/plate, CM881 and CM891 2 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:

TA1537 without metabolic activation 30 µg/plate

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA98 without metabolic activation 1 µg/plate

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene 2 µg/plate TA1535, 10 µg/plate CM881 and CM891

Remarks:

with metabolic activation

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA1537, TA98 and TA100 with metabolic activation 5 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

ACTIVATION: S9 mix contained S9 fraction 10%, MgCl, KCl, sodium orthophosphate buffer pH 7.4, glucose-6-phosphate and NADP. 0.5 ml S9 mix and 0.1ml of test solution were added to 2ml agar.

DURATION

- Preincubation period: 30 minutes
- Exposure duration: 3 days

SELECTION AGENT (mutation assays): histidine deficient agar

NUMBER OF REPLICATIONS: triplicate plates, experiment repeated

DETERMINATION OF CYTOTOXICITY
- Method: other: condition of background lawn

ACTIVATION
- Aroclor induced rat liver S9 was present at 10% in the S9 mix, which included NADP and glucolse-6-phosphate as cofactors. 0.5 ml S9 mix was added to 2 ml top agar, 0.1 ml test solution or control and 0.1 ml bacterial culture giving a final concentration of approximately 2%.

Evaluation criteria:

A result is considered positive if there is a three-fold increase in the number of revertants in TA1535, TA1537 and TA98 over the solvent control and a two-fold increase for other strains. This should occur in both experiments and show some evidence of a dose-relationship. The positive controls must cause at least a doubling of mean revertant colony numbers over the mean of the solvent control.

Statistics:

None in report

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

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

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

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

Test results 3

Key result

Species / strain:

E. coli, other: E. coli WP2 pKM101 (CM881)

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

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

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

Test results 5

Key result

Species / strain:

S. typhimurium TA 98

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

Test results 6

Key result

Species / strain:

S. typhimurium TA 100

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

Any other information on results incl. tables

Average number of revertants per plate (mean of three plates)

Treatment µg/plate	TA 1535		TA 1537		TA 98		TA 100		CM 881		CM 891
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
Solvent control	14	16	9	10	19	23	108	127	46	53	179	180
50	14	19	7	6	22	27	110	133	56	67	201	190
150	14	20	7	7	19	24	111	132	63	77	189	203
500	14	16	7	8	24	24	109	113	46	56	193	203
1500	13	16	6	7	25	23	111	123	61	73	199	195
5000	10	11	9	9	15	23	87	114	61	63	188	197
Positive control	370	179	316	46	115	164	456	718	817	392	1703	1799
Average Revertants per plate with and without metabolic activation – Exp 2 pre-incubation
Treatment µg/plate	TA 1535		TA 1537		TA 98		TA 100		CM 881		CM 891
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9		+S9
Solvent control	18	15	9	9	29	27	134	143	61	69	247		242
50	20	16	8	11	27	31	115	134	55	73	238		253
150	18	18	9	10	21	30	110	135	47	66	216		237
500	13	21	10	9	26	26	106	129	58	62	261		253
1500	18	18	7	8	21	28	113	114	54	80	230		235
5000	15	18	8	6	25	28	105	97	48	60	215		268
Positive control	244	115	120	144	130	270	456	569	792	438	1569		758

Average Revertants per plate with and without metabolic activation – Exp 1

Applicant's summary and conclusion

Conclusions:

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria, in a study that was conducted according to the OECD TG 471 and in compliance with GLP. No evidence of test substance related increase in the number of revertants was observed with or without activation in the initial experiment using the plate incorporation method or the repeat pre-incubation assay in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, E coli WP2 trp uvrA pKM101 (CM891) and E coli WP2 trp pKM101 (CM881). The solvent used was water, and hydrolysis was likely to have occurred under test conditions, so it is assumed that exposure was to the hydrolysis product as well as the parent substance. It is not considered to invalidate the study, as it is considered that hydrolysis may occur in vivo. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.