Bis(triethoxysilylpropyl)amine

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:

08 Sep - 02 Nov 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 1997

Deviations:

yes

Remarks:

2-AA was used as sole positive control in the presence of S9-mix, which is in conflict with the OECD guideline 471 (adopted 1997). No further information on S9 characterisation was given from the supplier.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis(triethoxysilylpropyl)amine

EC Number:

236-818-1

EC Name:

Bis(triethoxysilylpropyl)amine

Cas Number:

13497-18-2

Molecular formula:

C18H43NO6Si2

IUPAC Name:

bis(triethoxysilylpropyl)amine

Method

Target gene:

S. typhimurium strains: his operon
E. coli strain: trp 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 pKM 101

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone

Test concentrations with justification for top dose:

Dose Range Finder: 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate (±S9)
Main test:
-S9: 9.78, 19.6, 39.1, 78.3, 157, 313 µg/plate (TA 100); 39.1, 78.1, 156, 313, 625, 1250 µg/plate (TA 98, TA 1535, TA 1537, E. coli)
+S9: 39.1, 78.1, 156, 313, 625, 1250 µg/plate

Vehicle / solvent:

DMSO (Lot No: K40982531)
- Justification for choice of solvent/vehicle: Based on sponsor information and the result of a preliminary solubility test.

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

other: 2-aminoanthracene (2-AA), 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF2)

Remarks:

+S9: 2-AA (TA 98, TA 100, TA 1535, TA 1537, WP2uvrA; 1, 2, 3, 3, 2 µg/plate); -S9: 2-NF (TA 98; 5 µg/plate); SA (TA 100, TA 1535; 1.5 µg/plate); 9-AA (TA 1537; 80 µg/plate); AF2 (WP2uvrA; 0.005 µg/plate)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

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

DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition

Evaluation criteria:

The results were considered positive when the following conditions are met:
- The number of revertant colonies in the test substance groups was increased at least twice as compared to the negative control group at one or more doses per plate in at least one strain.
- The numer of revertant colonies was increased dose dependently.
- The results were reproducible.

Statistics:

Mean values and standard deviation were calculated

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

625 µg/plate (without S9); 1250 µg/plate (with S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

313 µg/plate (without S9); 1250 µg/plate (with S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1250 µg/plate (with and without S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1250 µg/plate (with and without S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1250 µg/plate (with and without S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Deposition was observed at 5000 µg/plate

RANGE-FINDING/SCREENING STUDIES: In the preliminary test growth inhibition was observed in all strains at 1250 µg/plate in the absence and presence of S9-mix. In addition growth inhibition was observed at 313 µg/plate in TA 100 in the absence of S9-mix.

Any other information on results incl. tables

Table 1: Test results of main test

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 (pKM101)	TA98	TA1537
–	Solvent control	73±2	12±2	104±7	10±1	5±1
–	9.78	69±1	-	-	-	-
–	19.6	71±1	-	-	-	-
–	39.1	64±7	7±5	106±1	12±1	5±1
–	78.1 (78.3, TA 100)	68±4	9±1	114±4	11±2	4±2
–	156 (157, TA 100)	66±4	8±1	132±3	11±3	5±0
–	313	15±15	9±2	99±15	11±3	6±4
–	625	-	7±3	124±16	9±2	3±2
–	1250	-	4±1	90±12	7±3	5±2
Positive controls, –S9	Name	sodium azide	sodium azide	AF2	2-nitrofluorene	9-amino-acridine
	Concentrations (μg/plate)	1.5	1.5	0.005	5	80
	Average of 3 plates±sd	294±36	425±8	905±250	550±79	642±90
+	Solvent control	81±5	8±3	128±13	17±4	12±
+	39.1	85±12	7±1	130±33	19±1	10±3
+	78.1	91±9	5±1	54±6	21±0	9±2
+	156	81±6	6±1	127±42	21±2	10±4
+	313	82±13	7±1	142±48	19±1	10±4
+	625	80±7	7±1	148±1	18±1	14±2
+	1250	59±8	5±1	107±18	12±3	8±3
Positive controls, +S9	Name	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene
	Concentrations (μg/plate)	2	3	2	1	3
	Average of 3 plates±sd	194±27	118±21	544±28	191±30	239±39

 

Table 2: Test results of main test 2.

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 (pKM101)	TA98	TA1537
–	Solvent control	72±5	8±2	114±8	12±3	5±3
–	9.78	69±15	-	-	-	-
–	19.6	70±1	-	-	-	-
–	39.1	65±3	10±2	119±2	10±4	4±2
–	78.1 (78.3, TA 100)	65±9	9±0	117±8	10±3	6±2
–	156 (157, TA 100)	66±8	10±2	102±1	12±4	5±3
–	313	49±8	8±3	137±7	10±3	4±1
–	625	-	6±2	130±8	10±5	4±1
–	1250	-	7±4	121±13	6±1	3±1
Positive controls, –S9	Name	sodium azide	sodium azide	AF2	2-nitrofluorene	9-amino-acridine
	Concentrations (μg/plate)	1.5	1.5	0.005	5	80
	Average of 3 plates±sd	349±38	420±15	917±15	376±34	313±20
+	Solvent control	85±11	8±3	149±19	16±7	11±4
+	39.1	69±3	8±2	136±8	14±6	10±
+	78.1	71±10	6±2	148±9	16±4	12±4
+	156	70±11	7±2	154±18	14±3	9±5
+	313	79±4	11±4	149±11	16±	13±1
+	625	79±3	7±2	156±10	20±7	11±6
+	1250	62±7	7±3	155±10	17±2	7±3
Positive controls, +S9	Name	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene	2-aminoanthracene
	Concentrations (μg/plate)	2	3	2	1	3
	Average of 3 plates±sd	298±25	99±7	482±31	150±13	171±3

 

 

Applicant's summary and conclusion

Conclusions:

negative

3-(Triethoxysilyl)-N-[3-(triethoxysilyl)propyl]-1-propanamine 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.