N-[3-(trimethoxysilyl)propyl]aniline

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Qualifier:

according 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]aniline

EC Number:

221-328-2

EC Name:

N-[3-(trimethoxysilyl)propyl]aniline

Cas Number:

3068-76-6

Molecular formula:

C12H21NO3Si

IUPAC Name:

N-[3-(trimethoxysilyl)propyl]aniline

Details on test material:

- Name of test material (as cited in study report): SILQUEST Y-9669 Silane
- Physical state: clear colourless liquid
- Analytical purity: 97.1%
- Lot/batch No.: 16984-17
- Storage condition of test material: original container at room temperature, protected from light

Method

Target gene:

his operon

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:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.

Test concentrations with justification for top dose:

preincubation test: 100, 333, 1000, 3333, 5000 µg/plate (experiment B1; valid for all strains)
preincubation test: 100, 333, 1000, 3333, 5000 µg/plate (experiment B3; valid for WP2 uvrA)
preincubation test: 3.3, 10, 33, 100, 333, 1000, 5000 µg/plate (experiment B4; valid for all Salmonella strains)

Vehicle / solvent:

Acetone

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Remarks:

+S9: 2-AA (all salmonella strains + WP2uvrA; 1 + 10 µg/plate); sterigmatocystin (WP2; 100 µg/plate); -S9: 2-NF (TA 98; 1 µg/plate); SA (TA 100, TA 1535; 1 µg/plate); 9-AA (TA 1537; 75 µg/plate); MMS (both E. coli strains; 1000 µg/plate)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 60 min
- Exposure duration: 48-72 h

NUMBER OF REPLICATIONS: 2 independent experiments with 3 plates/concentration.

DETERMINATION OF CYTOTOXICITY
- Method: reduction of bacterial background lawn; mean number of revertants per plate

Evaluation criteria:

Cytotoxicity:
- reduction of colony background lawn (code 1-5)
- reduction of mean number of revertants per plate as compared to the mean vehicle control value of >50%
Mutagenicity:
Positive reaction: dose-related increase in the mean revertants per plate of at least one tester strain with a minimum of two increasing concentrations of test article. Data sets for strains TA1535 and TA1537 were judged positive if the increase in mean revertants at the peak of the dose response is equal to or greater than three times the mean vehicle control value. Data sets for strains TA98, TA100 and WP2 uvrA (pKM101) and WP2 (pKM101) were judged positive if the increase in mean revertants at the peak of the dose response is equal to or greater than two times the mean vehicle control value.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: not observed up to 5000 µg/plate

RANGE-FINDING/SCREENING STUDIES: In the preliminary test cytotoxicity was not observed up to 5000 µg/plate

Any other information on results incl. tables

With or without S9-Mix	Test substance concentration	Mean number of revertant colonies per plate in the plate incorporation test
	(µg/plate)	(average of 3 plates±standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	WP2 uvrA	TA98	TA1537
–	Solvent control	225±6	7±3	13±2	13±3	3±1
–	100	232±9	7±3	15±1	14±2	4±1
–	333	220±18	8±3	14±1	13±3	5±3
–	1000	209±14	8±1	16±2	14±4	3±2
–	3333	215±23	6±5	11±3	11±1	4±3
–	5000	229±28	6±3	10±1	16±4	3±2
Positive controls, –S9	Name	sodium azide	sodium azide	methyl methane sulfonate	2-nitrofluorene	9-amino-acridine
	Concentrations (µg/plate)	1	1	1000	1	75
	Average of 3 plates±sd	678±17	399±38	445±215	289±54	623±36
+	Solvent control	234±2	7±3	13±2	14±3	3±1
+	100	272±10	7±3	15±1	16±3	4±1
+	333	272±9	8±3	14±1	16±5	5±3
+	1000	232±15	8±1	16±2	14±4	3±2
+	3333	212±12	6±5	11±3	24±2	4±3
+	5000	237±44	6±3	10±1	22±8	3±2
Positive controls, +S9	Name	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene
	Concentrations (µg/plate)	1	1	10	1	1
	Average of 3 plates±sd	1022±56	272±18	72±13	813±79	413±101

Table 1: Test results of main test B1.

 

Table 2: Test results of the repeat experiment B3 (WP2 uvrA) and B4 (all Salmonella strains).

With or without S9-Mix	Test substance concentration	Mean number of revertant colonies per plate in the plate incorporation test
	(µg/plate)	(average of 3 plates±standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	WP2 uvrA	TA98	TA1537
–	Solvent control	126±5	8±2	20±2	14±3	3±2
–	3.3	114±13	14±1	-	17±4	4±1
–	10	111±6	9±0	-	17±5	5±4
–	33	108±20	6±2	-	16±3	6±0
–	100	123±5	7±2	14±4	14±6	5±2
–	333	76±23	12±3	11±3	16±5	4±1
–	1000	104±13	6±3	12±4	16±7	4±2
–	3333	-	-	12±1	-	-
–	5000	594±5	7±4	13±3	17±3	4±3
Positive controls, –S9	Name	sodium azide	sodium azide	methyl methane sulfonate	2-nitrofluorene	9-amino-acridine
	Concentrations (µg/plate)	1	1	1000	1	75
	Average of 3 plates±sd	594±5	657±69	363±4	260±20	52±18
+	Solvent control	129±8	10±2	16±5	22±1	7±0
+	3.3	138±13	10±3	-	19±2	8±5
+	10	134±10	11±6	-	28±6	5±2
+	33	130±8	11±4	-	22±2	4±1
+	100	138±8	8±4	14±6	20±7	6±2
+	333	143±12	9±1	17±2	25±7	5±2
+	1000	53±19	8±2	14±0	22±7	3±3
+	3333	-	-	17±5	-	-
+	5000	131±22	8±3	14±2	24±6	7±4
Positive controls, +S9	Name	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene
	Concentrations (µg/plate)	1	1	10	1	1
	Average of 3 plates±sd	858±31	94±6	106±19	731±15	97±23

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative

N-[3-(trimethoxysilyl)propyl]aniline has been tested for mutagenicity in bacteria, in a study which was conducted according to OECD 471 and 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 a preincubation experiment. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.