Reaction mass of 2-{[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and 2-{benzyl[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and N-[2-(benzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride and N-[2-(dibenzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride and N-benzyl-2-{benzyl[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and N-benzyl-N-[2-(dibenzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride.

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:

22 June - 06 July 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The study was conducted according to OECD TG 471 with acceptable restrictions. The restriction was that no additional strain for the detection of crosslinking agents was included into the study. The study was further conducted in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(1983)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(1997)

Deviations:

yes

Remarks:

(no additional strain for the detection of crosslinking agents was included into the study)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: EU Method B.14 (Mutagenicity - Reverse Mutation Test Using Bacteria) (1984)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2-{[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and 2-{benzyl[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and N-[2-(benzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride and N-[2-(dibenzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride and N-benzyl-2-{benzyl[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and N-benzyl-N-[2-(dibenzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride.

Molecular formula:

Not applicable

IUPAC Name:

Reaction mass of 2-{[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and 2-{benzyl[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and N-[2-(benzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride and N-[2-(dibenzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride and N-benzyl-2-{benzyl[3-(trimethoxysilyl)propyl]amino}ethanaminium chloride and N-benzyl-N-[2-(dibenzylamino)ethyl]-3-(trimethoxysilyl)propan-1-aminium chloride.

Test material form:

liquid: viscous

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

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

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:

- experiment 1: 3.3, 10, 33, 100, 200 µg/plate
- experiment 2: 3.3, 10, 33, 100, 333 µg/plate

Vehicle / solvent:

- Solvent used: DMSO (spectroscopic quality (Merck) to which molecular sieve (0.4 nm) beads (about 2 mm) were added to absorb the water)
- Justification for choice of solvent: The test article was found to be not stable in water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: sodium azide (SA; 1 µg/plate;TA1535); 9-aminoacridine (9AC; 60 µg/plate; TA1537); daunomycin (DM; 4 µg/plate; TA98); methylmethanesulfonate (MMS; 650 µg/plate; TA100);+S9: 2-aminoanthracene (2AA; 5 µg/plate:TA1535 and TA1537;0.5 µg/plate:TA98/TA100)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: none
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replicates in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth (In a preliminary cytotoxicity experiment with TA100 both in absence and presence of metabolic activation nine concentrations of the test item were tested for cytotoxicity. The highest concentration of the test article used in the subsequent mutagenesis assay was that which gave a reduced survival on the non-selective plates.)

Evaluation criteria:

A test substance is considered negative (not mutagenic) in the Ames test if:
- The total number of revertants in any tester strain at any concentration is not greater than two times the solvent control value, with or without metabolic activation.
- The negative response should be reproducible in at least one independently repeated experiment.

A test substance is considerd positive (mutagenic) in the Ames test if:
- It induces at leats a 2-fold, dose related increase in the number of revertants with respect to the number induced by the solvent control in any of the tester strains, either with or without metabolic activation. However, any mean plate count of less than 20 is considered to be not significant. If the test substance showed in the first test only a positive response at one or two concentrations, the assay was repeated with doses below and exceeding those showing positive effects in the first test.
- The positive response should be reproducible in at least one independently repeated experiment.

The preceding criteria were not absolute and other modifying factors might enter into the final evaluation decision.

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:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- none

ADDITIONAL INFORMATION ON CYTOTOXICITY: The cytotoxicity of the test item was tested with TA100 in absence and presence of metabolic activation. In the presence of S9-mix the survival of strain TA100 is not reduced up to and including test substance concentrations of 33 µg/plate, slightly reduced at a concentration fo 100 µg/plate, extremely reduced at concentrations from 333 µg/plate up to and including 3330 µg/plate and absent at a concentration of 5000 µg/plate.
In the absence of S9-mix the survival of strain TA100 is not reduced up to and including test substance concentrations of 33 µg/plate, moderately reduced at a concentration fo 100 µg/plate, extremely reduced at a concentrations of 333 µg/plate and absent at test substance concentrations of 1000 µg/plate and upwards.
Based on these data, the test substance was tested up to and including a concentration of 200 µg/plate in the absence and presence of metabolic activation (experiment 1). In the first experiment no toxicity was observed, therefore it was decided to test up to and including a concentration of 333 µg/plate in the second experiment.

Any other information on results incl. tables

Tab. 1: Mutagenic response of ORGANOFUNCTIONAL SILANE A-1128 in the Ames Salmonella/microsome plate test. Given is the mean number of revertant (His+) colonies / 3 replicate plates (± SD) with different strains of S. typhimurium (experiment 1).

Dose [µg/plate]		TA1535		TA1537		TA100		TA98
		+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9
Test item	3.3	17 ± 5	16 ± 1	7 ± 3	6 ± 2	29 ± 6	21 ± 1	97 ± 7	88 ± 8
	10	20 ± 5	17 ± 3	6 ± 1	3 ± 2	27 ± 3	20 ± 2	97 ± 4	85 ± 10
	33	15 ± 5	15 ± 4	6 ± 1	4 ± 2	28 ± 1	21 ± 5	105 ± 4	85 ± 12
	100	16 ± 2	15 ± 6	9 ± 2	6 ± 1	29± 6	25 ± 3	112 ± 6	87 ± 1
	200	12 ± 3	9 ± 2	9 ± 1	6 ± 2	26 ± 6	22 ± 2	113 ± 6	75 ± 21
Solvent control		14 ± 1	19 ± 1	11 ± 1	4 ± 2	30 ± 5	22 ± 4	87 ± 6	89 ± 8
Positive control		282 ± 49	273 ± 15	902 ± 64	154 ± 22	362 ± 25	321 ± 34	585 ± 71	844 ± 99

 

Tab. 2: Mutagenic response of ORGANOFUNCTIONAL SILANE A-1128 in the Ames Salmonella/microsome plate test. Given is the mean number of revertant (His+) colonies / 3 replicate plates (± SD) with different strains of S. typhimurium (experiment 2).

Dose [µg/plate]		TA1535		TA1537		TA100		TA98
		+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9
Test item	3.3	7 ± 2	9 ± 3	8 ± 3	6 ± 1	33 ± 2	32 ± 4	88 ± 7	80 ± 8
	10	7 ± 1	10 ± 4	8 ± 5	6 ± 3	30 ± 3	25 ± 3	84 ± 5	69 ± 3
	33	11 ± 2	9 ± 2	8 ± 3	5 ± 2	30 ± 8	22 ± 7	84 ± 8	63 ± 20
	100	11 ± 3	13 ± 1	10 ± 6	7 ± 1	32± 1	28 ± 6	100 ± 19	84 ± 10
	333	12 ± 2	6 ± 5	5 ± 2	3 ± 1	25 ± 5	19 ± 9	70 ± 2	40 ± 8
Solvent control		10 ± 1	8 ± 1	6 ± 2	7 ± 2	30 ± 6	27 ± 4	92 ± 26	80 ± 10
Positive control		252 ± 59	198 ± 28	1008 ± 10	145 ± 7	493 ± 20	443 ± 68	576 ± 13	1030 ± 18

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative with and without metabolic activation

The test item was tested for mutagenicity to bacteria according to the OECD TG 471 (1983) and in compliance with GLP. No additional strain for the detection of crosslinking agents was included into the study. No test material related increase in revertants was observed up to cytotoxic concentrations. Hence, the test item is not mutagenic under the conditions of the study.