N,N,N',N'-tetramethyl-N''-[3-(trimethoxysilyl)propyl]guanidine

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:

2015-11-26 to 2015-12-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N,N',N'-tetramethyl-N''-[3-(trimethoxysilyl)propyl]guanidine

EC Number:

274-092-8

EC Name:

N,N,N',N'-tetramethyl-N''-[3-(trimethoxysilyl)propyl]guanidine

Cas Number:

69709-01-9

Molecular formula:

C11H27N3O3Si

IUPAC Name:

N,N,N',N'-tetramethyl-N''-[3-(trimethoxysilyl)propyl]guanidine

Test material form:

other: liquid

Method

Target gene:

histidine operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

phenobarbital (80 mg/kg bw) and β-naphthoflavone (100 mg/kg bw) induced rat liver S9

Test concentrations with justification for top dose:

0.0316, 0.100, 0.316, 1.0, 2.5 and 5.0 μL/plate

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 the S9 activity

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

treatment medium

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100 and TA 1535, without metabolic activation

Controls 2

Untreated negative controls:

yes

Remarks:

treatment medium

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Remarks:

TA 98 and TA 1537, without metabolic activation

Controls 3

Untreated negative controls:

yes

Remarks:

treatment medium

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA 102, without metabolic activation

Controls 4

Untreated negative controls:

yes

Remarks:

treatment medium

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

TA 98, TA 100, TA 1535, TA 1537 and TA 102, with metabolic activation

Details on test system and experimental conditions:

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

ACTIVATION: The S9 mix contained 9.5 parts co-factor solution and 0.5 parts liver preparation. The co-factor solution consisted of 8 mM MgCl2, 33 mM KCl, 5 mM glucose-6-phosphate, 4 mM NADP in 100 mM of sodium-ortho-phosphate-buffer, pH 7.4. The final protein concentration in the S9 preparation was 30.1 mg/mL.

DURATION
- Preincubation period: 60 min at 37°C
- Exposure duration: 48 hours in the dark

SELECTION AGENT (mutation assays):

NUMBER OF REPLICATIONS: triplicate cultures

DETERMINATION OF CYTOTOXICITY
- Method: background lawn, number of revertant colonies

OTHER EXAMINATIONS:
-precipitation

Evaluation criteria:

A test item is considered as mutagenic if:
- a clear and dose-related increase in the number of revertants occurs and/or
- a biologically relevant positive response for at least one of the dose groups occurs in at least one tester strain with or without metabolic activation.

Statistics:

Mean value and statistical significance were calculated.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Experiement II: TA 98, TA 100 and TA 1535 at 5.0 μL/plate (-S9), and 2.5 μL/plate and higher (+S9); TA 1537 and TA 102 at 2.5 μL/plate and higher (+/- S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitation of the test item was observed in experiment I and II, with and without metabolic activation

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Experiment I: no cytotoxic effect was noted in any of the test strains at any of the test doses, with and without metabolic activation.
- Experiment II: cytotoxic effect of the test item was noted in test strains TA 98, TA 100 and TA 1535 at 5.0 μL/plate, without metabolic activation, and at 2.5 μL/plate and higher, with metabolic activation. In test strains TA 1537 and TA 102 toxic effect was noted at concentration 2.5 μL/plate and higher, with and without metabolic activation.

Any other information on results incl. tables

Table 1: Experiment I, mean number of revertants, with and without metabolic activation

Concentration µL/plate	TA 98		TA 100		TA 1535		TA 1537		TA 102
	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA
Negative control	41	31	78	78	8	10	7	8	417	230
DMSO	34	31	73	75	7	6	5	6	352	208
0.0316	37	38	99	76	8	11	10	8	393	268
0.100	38	34	81	74	9	12	9	7	362	228
0.316	35	35	94	76	5	9	6	4	346	228
1.0	38	33	74	71	6	7	8	7	302	219
2.5	39	35	70	67	7	12	8	10	319	235
5.0	40	23	81	62	11	12	8	10	362	272
4-NOPD 10 μg	-	361	-	-	-	757	-	-	-	-
4-NOPD 40 μg	-	-	-	-	-	-	-	86	-	-
NaN3 10 μg	-	-	-	528	-	-	-	-	-	-
MMS 1 μL	-	-	-	-	-	-	-	-	-	1084
2-AA 2.5 μg	2335	-	1255	-	180	-	338	-	-	-
2-AA 10 μg	-	-	-	-	-	-	-	-	882	-

Table 2: Experiement II, mean number of revertants, with and without metabolic activation

Concentration µL/plate	TA 98		TA 100		TA 1535		TA 1537		TA 102
	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA
Negative control	41	39	100	88	11	9	10	6	384	339
DMSO	39	30	77	74	10	11	9	4	292	296
0.0316	40	23	77	80	10	9	11	8	376	266
0.100	39	29	80	68	7	10	9	7	387	284
0.316	37	34	87	78	9	8	8	5	351	233
1.0	40	33	80	86	10	10	8	8	360	236
2.5	31	30	67	70	8	11	7	5	283	231
5.0	32	27	55	0	8	0	8	6	288	241
4-NOPD 10 μg	-	459	-	-	-	-	-	-	-	-
4-NOPD 40 μg	-	-	-	-	-	-	-	103	-	-
NaN3 10 μg	-	-	-	770	-	1019	-	-	-	-
MMS 1 μL	-	-	-	-	-	-	-	-	-	2382
2-AA 2.5 μg	1153	-	1132	-	81	-	104	-	-	-
2-AA 10 μg	-	-	-	-	-	-	-	-	599	-

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative with and without metabolic activation

1,1,3,3-tetramethyl-2-[3-(trimethoxysilyl)propyl]guanidine has been tested in a valid bacterial mutation assay, conducted according to OECD TG 471, and in compliance with GLP, using S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102. No increase in the number of revertants was observed in any of the five test strains, with and without metabolic activation, when tested up to cytotoxic concentration. Appropriate solvent, negative and positive 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.